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Color Prove Manual MK

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Color Prove Manual MK

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marco.hang00

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Manual

Methods for color

measurements

S
pectroquant ®

Prove
Spectroquant® Prove - Methods for color measurements

2 Release 10/2020
Spectroquant® Prove - Methods for color measurements

Contents
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . 5
I Colorimetry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

II Overview of methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

III Typical areas of application . . . . . . . . . . . . . . . . . . . . . . . 10

Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
1 ADMI color measurement . . . . . . . . . . . . . . . . . . . . . . . . 14

2 Anisidine value . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

3 ASTM color measurement . . . . . . . . . . . . . . . . . . . . . . . . 24

4 CIE color distance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

5 CIELAB color space (brightness, chroma) . . . . . . . . . . . . 31

6 CIELUV color space . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34

7 CIExyY color space . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

8 Color (ASBC method) . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

9 Color (EBC method) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47

10 Color Hazen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52

11 Color 410 acc. to EN 7887 . . . . . . . . . . . . . . . . . . . . . . . . 57

12 Color – Spectral absorption coefficient

acc. to DIN EN ISO 7887 . . . . . . . . . . . . . . . . . . . . . . . . . 61

13 Gardner color measurement . . . . . . . . . . . . . . . . . . . . . . 68

14 Hess-Ives color scale . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71

15 Hunter color distance . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74

16 HunterLab color space . . . . . . . . . . . . . . . . . . . . . . . . . . . 78

17 ICUMSA Color GS1/3-7 . . . . . . . . . . . . . . . . . . . . . . . . . . 81

18 ICUMSA Color GS2/3-9 . . . . . . . . . . . . . . . . . . . . . . . . . . 85

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Methods
19 ICUMSA Color GS2/3-10 . . . . . . . . . . . . . . . . . . . . . . . . . 89

20 ICUMSA Color GS9/1/2/3-8 . . . . . . . . . . . . . . . . . . . . . . 93

21 Iodine color number . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98

22 Klett color index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103

23 Saybolt color measurement . . . . . . . . . . . . . . . . . . . . . . . 107

24 Spectral absorption coefficient, SAC α(254)* . . . . . . . . . 110

25 Spectral absorption coefficient, SAC α(436) . . . . . . . . . . 114

26 Spectral attenuation coefficient, SAC µ(254)* . . . . . . . . 117

27 Spectral attenuation coefficient, SAC µ(254) -

corrected* . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121

28 Tint index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125

29 Transmittances TX, TY, TZ . . . . . . . . . . . . . . . . . . . . . . . . . 128

30 UV-absorbing organic matter (UV absorption 254)* . . . . 132

31 UV irradiation (UV absorption 254 -

UV transmission 254)* . . . . . . . . . . . . . . . . . . . . . . . . . . 137

32 Whiteness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 144

33 Yellowness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147

List of smart icons on display . . . . . . . . . . . .150

* not for Spectroquant® Prove 100

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Introduction
I Colorimetry
The perception of color and its interpretation play an essential role in everyday life. In nature, for example,
red and yellow count as warning colors. With their bright yellow or vivid red appearance, poison-dart frogs
scare off their potential enemies, thus securing their survival and at the same time signalling their toxicity.
The effect of these colors is used to communicate messages in many areas of everyday life, for instance in
the form of signalling systems, mandatory instructions, and warning signs. In the food area, the colors can
provide valuable information on the maturity of the product. Whenever products like paper or drinking water
are affected, discolorations are an andesirable effect and are taken as an indication of contamination or
impurity. In other applications, however, a comparison of colors plays a major role. These include, for
instance, the assessment of the quality of beer, surfaces, printing inks, paints, and oils, the analysis of
printed materials – or using color scales to assess detection reactions, e. g. in the determination of the pH of
products using pH test strips.
In the easiest cases, the human eye is the ideal tool for the assessment of a color. The differences in per-
ception of colors between one person and the next, however, can result in considerable variances in the final
interpretation. This is why color scales or color charts, colored solutions, or color glasses are used in many
cases to facilitate an objective color comparison or color match. The most important color scales used in this
regard are the Ph Eur and the US Pharmacopoeia color scales, the Hazen or Pt/Co color scale, and the
iodine color value. Beyond these, colors can be compared visually using color systems such as the RAL® or
Pantone® systems, for which there are a broad variety of sample books and charts as well as catalogues
depicting innumerable color nuances covering the entire visual color spectrum.
One weakness of the visual method for comparing colors – besides the influence of the individual observer –
is the impact of other factors, with the ambient light, for instance, or the angle of observation, or the dis-
tance between the observer and the object – all potentially affecting the way in which the color is perceived.
As a measure to prevent these – in some cases subjective – factors from distorting the objective results,
many scientists resort to spectral methods using state-of-the-art measurement instruments. These devices
run a wavelength scan over the entire visual spectrum (approx. 360 nm to 800 nm) or measure at specific
wavelengths, determining the reflection or transmission or else the absorption of the sample ander investi-
gation. The results can then be used to calculate the most varied color spaces (e. g. LAB, LUV), color values
(e. g. Gardner, Saybolt), or fine color gradations (e. g. EBC, ICUMSA).
The precise conditions for measurement and the principle by which results are calculated are described in
the individual method descriptions of this manual or in the cited technical Literature.

Note
The measurement conditions and the calculation formulae of the colorimetric methods are pre-programmed
in the Prove spectrophotometers. After selecting the method, the device menu guides the user through the
procedure. The measurement results are automatically calculated and shown in the spectrophotometer dis-
play.

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II Overview of methods
Parameter Standard Sample material Measuring range Method principle Method No. Photometer Prove
ADMI color APHA 2120 F Liquid samples 2.0 - 100.0 ADMI Transmittances from 2518 100, 300, 600
measurement 10 - 600 ADMI 400 - 700 nm 2517
10 - 1000 ADMI 2516
Anisidine value ISO 6885, Animal and vegeta- 0.0 - 200.0 AV Color comparison at 2584 100, 300, 600
DGF C-VI 6e ble fats and oils 350 nm
ASTM color ASTM D6045 Liquid samples, 0.5 - 8.0 ASTM Transmittances from 2562 100, 300, 600
measurement petroleum products 380 - 780 nm

CIE color DIN EN 11664-4, Liquid samples ΔE*ab 0.00 - 200.00 Comparative measure- 2584 100, 300, 600
distance DIN 5033-3 ΔL* -200.00 - 200.00 ment of transmittances
Δa*-200.00 - 200.00 from 360 - 780 nm
Δb*-200.00 - 200.00
ΔC*ab -200.00 - 200.00
CIELAB color DIN EN 11664-4, Liquid samples ΔE*ab 0.00 - 200.00 Comparative measure- 2584 100, 300, 600
space (bright- DIN 5033-3 ΔL* -200.00 - 200.00 ment of transmittances
ness, chroma) Δa*-200.00 - 200.00 from 360 - 780 nm
Δb*-200.00 - 200.00
ΔC*ab -200.00 - 200.00
CIELUV color DIN 5033-3, Liquid samples L* 0.00 - 105.00 Transmittances from 2581 100, 300, 600
space ISO CIE 11664-5 u* -180.0 - 180.0 360 - 780 nm
v* -180.0 - 180.0
C*uv 0.00 - 300.00
S*uv 0.000 - 200.000
CIExyY color CIE 15:2004, Liquid samples x 0.0000 - 0.8000 Transmittances from 2582 100, 300, 600
space Technical Report y 0.0000 - 0.8000 360 - 780 nm
- Colorimetry Y 0.000 - 200.000
Color (ASBC ASBC Method Beers, worts, 0.0 - 50.0 °SRM Absorption at 430 nm 2633 100, 300, 600
method) Beer-10, liquid malt substi- 0.0 - 100.0 EBC Units
ASBC Method tutes
Wort-9
Color (EBC MEBAK Method Beers, worts, 0.0 - 60.0 °SRM Absorption at 430 nm 2602 100, 300, 600
method) 2.13.2, liquid malt substi-
EBC Method 8.5 tutes
and 9.6
Color Hazen
340 nm - Yellow to yellow- 0.2 - 500 CU Absorption at 340 nm 32 100, 300, 600
brownish liquid sam-
ples
445 nm DIN EN ISO Yellow to yellow- 1 - 1000 CU Absorption at 445 nm 179 100, 300, 600
6271, brownish liquid sam-
ASTM D 1209-05 ples
455 nm DIN EN ISO Yellow to yellow- 1 - 1000 CU Absorption at 455 nm 180 100, 300, 600
6271, brownish liquid sam-
APHA 2120C, ples
ASTM D 1209-05
465 nm DIN EN ISO Yellow to yellow- 1 - 1000 CU Absorption at 465 nm 181 100, 300, 600
6271, brownish liquid sam-
APHA 2120C, ples
ASTM D 1209-05
Color 410 acc. to DIN EN ISO Water from water- 2 - 2500 CU Absorption at 410 nm 303 100, 300, 600
EN 7887 7887:2011 treatment plants
Verfahren C

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Parameter Standard Sample material Measuring range Method principle Method No. Photometer Prove
Color – Spectral
absorption coef-
ficient acc. to
DIN EN ISO 7887

436 nm DIN EN ISO Water from water- 0.1 - 250.0 m-1 Absorption at 436 nm 15 100, 300, 600
7887:2011 - treatment plants
Verfahren B,
deutsche Trink-
wasserverord-
nung Anlage 3,
No. 7
SAK α(436) DIN EN ISO Water from water- 0.1 - 250.0 m-1 Absorption at 436 nm 302 100, 300, 600
7887:2011 - treatment plants
Verfahren B,
deutsche Trink-
wasserverord-
nung Anlage 3,
No. 7
525 nm DIN EN ISO Water from water- 0.1 - 250.0 m-1 Absorption at 525 nm 61 100, 300, 600
7887:2011 - treatment plants
Verfahren B
620 nm DIN EN ISO Water from water- 0.1 - 250.0 m-1 Absorption at 620 nm 78 100, 300, 600
7887:2011 - treatment plants
Verfahren B
436, 525 and DIN EN ISO Water from water- 0.0 - 250.0 m-1 Absorption at 436, 525 2588 100, 300, 600
620 nm 7887:2011 - treatment plants and 620 nm
Verfahren B
Gardner color DIN EN ISO Clear, yellow to yel- 1.0 - 18.0 Gardner Transmittances from 2561 100, 300, 600
measurement 4630-2, low-brownish liquid 360 - 780 nm
ASTM D6166 samples,
products made of
natural resins
Hess-Ives color DGK Prüfme- Fat derivatives, sur- 0.0 - 400 H-I Absorption at 460, 470, 2586 100, 300, 600
scale thode F 050.2 factants 560 and 640 nm
Hunter color HunterLab Liquid samples ΔE*H 0.00 - 200.00 Transmittances from 2585 100, 300, 600
distance Application Note ΔL* -200.00 - 200.00 360 - 780 nm
Vol. 8, No. 9, Δa*-200.00 - 200.00
06/08 Δb*-200.00 - 200.00
HunterLab color HunterLab Liquid samples L* 0.00 - 105.00 Transmittances from 2583 100, 300, 600
space Application Note a* -180.0 - 180.0 360 - 780 nm
Vol. 8, No. 9, b* -180.0 - 180.0
06/08
ICUMSA Color ICUMSA Method Sugar with a color 0 - 50 000 IU7.0 Absorption at 420 nm, 2548 100, 300, 600
GS1/3-7 Book GS1/3-7 factor >250 IU7.0 sample adjusted to pH 7
(raw sugar, strongly
colored white sugar
from plantations,
partly refined brown
sugar, sugar syrup)
ICUMSA Color ICUMSA Method Sugar with a color 0 - 600 IU7.0 Absorption at 420 nm, 2549 100, 300, 600
GS2/3-9 Book GS2/3-9 factor up to 600 IU7.0 sample adjusted to pH 7
(crystalline white
sugar, icing sugar,
sugar syrup)

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II Overview of methods
Parameter Standard Sample material Measuring range Method principle Method No. Photometer Prove
ICUMSA Color ICUMSA Method Sugar with a color 0 - 50 IU Absorption at 420 nm 2550 100, 300, 600
GS2/3-10 Book GS2/3-10 factor up to 50 IU
(crystalline white
sugar, icing sugar,
sugar syrup)
ICUMSA Color ICUMSA Method Sugar with a color 0 - 20 000 IU7.0 Absorption at 420 nm, 2551 100, 300, 600
GS9/1/2/3-8 Book factor up to 16000 sample adjusted to pH 7
GS9/1/2/3-8 IU7.0 (raw sugar,
white sugar from
plantations, refined
raw sugar)
Iodine color
number
445 nm DIN 6162, Yellow to yellow- 0.2 - 50.0 Absorption at 445 nm 21 100, 300, 600
DGF C-IV 4 a, brown liquid samples
F-I 2 and H-II 3
340 nm DIN 6162, Yellow to yellow- 0.010 - 3.00 Absorption at 340 nm 33 100, 300, 600
DGF C-IV 4 a, brown liquid sam-
F-I 2 and DGF ples, preferentially
H-II 3 for weakly colored
samples
Klett color index - Clear, yellow to yel- 0 - 1000 Klett417 Absorption at 417 nm 311 100, 300, 600
low-brown liquids
Saybolt color ASTM D6045 Liquid samples, -16.0 - 31.0 Saybolt Transmittances from 2563 100, 300, 600
measurement Petroleum products (50-mm cell) 380 - 780 nm 2564
-16.0 - 31.0 Saybolt
(100-mm cell)
Spectral absorp-
tion coefficient
254 nm DIN 38404-3 Water from water- 0.1 - 250 m-1 Absorption at 254 nm 300 300, 600
(SAC α(254)) treatment plants, filtered sample
surface water
436 nm DIN EN ISO Water from water- 0.1 - 250 m-1 Absorption at 436 nm 302 100, 300, 600
(SAC α(436)) 7887 - treatment plants filtered sample
Verfahren B,
deutsche Trink-
wasserverord-
nung Anlage 3,
No. 7,
EU-Trinkwasser-
richtlinie
98/83/EG
Spectral
attenuation
coefficient
254 nm
254 nm DIN 38404-3 Water from water- 0.1 - 250 m-1 Absorption at 254 nm, 301 300, 600
(SAC μ(254)) treatment plants, unfiltered sample
surface water
254 nm DIN 38404-3 Uncolored water 0.0 - 250 m-1 Absorption at 254 and 2571 300, 600
(SAC μ(254)), from water-treat- 550 nm, unfiltered,
corrected ment plants, uncol- uncolored sample
ored surface water
Tint index ASTM E313-15 Liquid samples -6.00 - 3.00 TI10mm Transmittances from 2577 100, 300, 600
(10-mm cell) 380 - 780 nm 2578
-6.00 - 3.00 TI50mm
(50-mm cell)
Transmittances DIN EN 1557 Liquid samples TX 0.0 - 150.0 Transmittances from 2579 100, 300, 600
TX, TY, TZ TY 0.0 - 150.0 380 - 780 nm
TZ 0.0 - 150.0

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Parameter Standard Sample material Measuring range Method principle Method No. Photometer Prove
UV-absorbing APHA 5910 Water from water- 0.0000 - 1.000 A/cm Absorption at 254 nm 309 300, 600
organic matter treatment plants, 0.0000 - 1.000 cm-1 filtered sample
surface water, 0.0 - 100 mm-1
seawater
UV absorption APHA 5910 Water from water- 0.0000 - 3.000 A/cm Absorption at 254 nm 310 300, 600
254 nm treatment plants, 0.0000 - 3.000 cm-1 filtered sample
surface water, 0.00 - 300.0 m-1
seawater
UV irradiation

UV absorption - Water from water- 0.0000 - 3.000 A/cm Absorption at 254 nm 310 300, 600
254 nm treatment plants, 0.0000 - 3.000 cm-1 unfiltered sample
surface water, 0.0 - 300.0 m-1
seawater
UV- tansmis- - Water from water- 0.00 - 105.00 %T/cm Absorption at 254 nm 2572 300, 600
sion 254 nm treatment plants, unfiltered sample
surface water,
seawater
Whiteness ASTM E313-15 Liquid samples 40.0 - 220.0 WI10mm Transmittances from 2575 100, 300, 600
(10-mm cell) 380 - 780 nm 2576
40.0 - 220.0 WI50mm
(50-mm cell)
Yellowness ASTM E313-15 Colorless to yellowish 0.0 - 30.0 YI10mm Transmittances from 2573 100, 300, 600
liquid samples (10-mm cell) 380 - 780 nm 2574
0.0 - 90.0 YI50mm
(50-mm cell)

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III Typical areas of application

ts
uc
od
pr
r

r
ga

he
er

Su
Be

Ot
Food and beverage
Parameter Samples
ADMI color measurement Liquid samples
Anisidine value Animal and vegetable fats and oils
ASTM color measurement Liquid samples, petroleum products
CIE color distance Liquid samples X
CIELAB color space (brightness, chroma) Liquid samples X
CIELUV color space Liquid samples X
CIExyY color space Liquid samples X
Color (ASBC method) Beers, worts, liquid malt substitutes X
Color (EBC method) Beers, worts, liquid malt substitutes X
Color Hazen
340 nm Yellow to yellow-brownish liquid samples
445 nm Yellow to yellow-brownish liquid samples
455 nm Yellow to yellow-brownish liquid samples
465 nm Yellow to yellow-brownish liquid samples
Color 410 acc. to EN 7887 Water from water-treatment plants
Color – Spectral absorption coefficient
acc. to DIN EN ISO 7887
436 nm Water from water-treatment plants
SAK α(436) Water from water-treatment plants
525 nm Water from water-treatment plants
620 nm Water from water-treatment plants
EN 7887 Water from water-treatment plants
Gardner color measurement Clear, yellow to yellow-brownish liquid samples,
products made of natural resins
Hess-Ives color scale Fat derivatives, surfactants
Hunter color distance Liquid samples X
HunterLab color space Liquid samples X
ICUMSA Color GS1/3-7 Sugar with a color factor >250 IU7.0
(raw sugar, strongly colored white sugar from plan- X
tations, partly refined brown sugar, sugar syrup)
ICUMSA Color GS2/3-9 Sugar with a color factor up to 600 IU7.0
X
(crystalline white sugar, icing sugar, sugar syrup)
ICUMSA Color GS2/3-10 Sugar with a color factor up to 50 IU
X
(crystalline white sugar, icing sugar, sugar syrup)
ICUMSA Color GS9/1/2/3-8 Sugar with a color factor up to 16000 IU7.0
(raw sugar, white sugar from plantations, refined X
raw sugar)
Iodine color number
445 nm Yellow to yellow-brown liquid samples
340 nm Yellow to yellow-brown liquid samples,
preferentially for weakly colored samples
Klett color index Clear, yellow to yellow-brown liquids

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III Typical areas of application

ts
uc
od
pr
r

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ga

he
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Su
Be

Ot
Food and beverage
Parameter Samples
Saybolt color measurement Liquid samples, petroleum products
Spectral absorption coefficient
254 nm (SAC α(254)) Water from water-treatment plants,
surface water
436 nm (SAC α(436)) Water from water-treatment plants
Spectral attenuation coefficient
254 nm (SAC μ(254))" Water from water-treatment plants,
surface water
254 nm (SAC μ(254)), korrigiert" Uncolored water from water-treatment plants,
uncolored surface water
Tint index Liquid samples
Transmittances TX, TY, TZ Liquid samples
UV-absorbing organic matter Water from water-treatment plants,
surface water, seawater
UVabsorption 254 nm Water from water-treatment plants,
surface water, seawater
UV irradiation Water from water-treatment plants,
surface water, seawater
UV absorption 254 nm Water from water-treatment plants,
surface water, seawater
UV transmission 254 nm Water from water-treatment plants,
surface water, seawater
Whiteness Liquid samples
Yellowness Colorless to yellowish liquid samples

12 Release 10/2020
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13
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Spectroquant® Prove - Methods for color measurements
Spectroquant® Prove - Methods for color measurements

Methods
1 ADMI color measurement
1.1 Method

The ADMI color scale originates from the “American Dye Manufacturers Institute” (ADMI) and is defined,
just like the Pt/Co color scale, by the color of hexachloroplatinate solutions.
The ADMI value is determined by measuring the transmittances (transmittances) in the wavelength spec-
trum between 400 and 700 nm and subsequent calculation according to Allen (see literature reference1).
Since the last step in the calculation of the ADMI value is based on the ΔE value of the sample relative to
water, this method can also be used to determine ADMI values for colors that lie beyond the yellow-orange
range of the Pt/Co scale.
The method is analogous to the APHA 2120 F standard (see literature reference2).

1.2 Measuring range

Method 2518 ADMI 50 2.0 - 100.0 ADMI

(50-mm rectangular cell)

Method 2517 ADMI 10 10 - 600 ADMI

(10-mm rectangular cell)

Method 2518 ADMI 10 10 - 1000 ADMI

(10-mm rectangular cell)

1.3 Sample material

Clear liquid samples

Turbid liquid samples after filtration

1.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114946 - Rectangular cells 10 mm and/or
Cat. No. 114944 - Rectangular cells 50 mm
ater for analysis EMSURE® or
• Cat. No. 116754 - W
distilled water
• Cat. No. 100716 - Sulfuric acid 25% for analysis EMSURE® (optional)
• Cat. No. 105588 - Sodium hydroxide solution min. 10% for analysis EMSURE® (optional)
• Cat. No. 100246 - Platinum Cobalt Color Reference Solution (HAZEN 500) with 500 mg/l Pt (optional)
• Membrane filters, pore size max. 0.45 µm (optional)

1.5 Preparation

• Filter turbid sample solutions over a membrane filter.

To do this, prerinse the membrane filter with approx. 50 ml of water for analysis, then rerinse with
approx. 50 ml of the sample solution and discard the filtrate.
Filter a further 50 ml of the sample solution over the prerinsed membrane filter and use the filtrate for
the analysis.
• Determination at the original pH: Filter turbid samples if necessary.
• Determination at pH 7.0: Filter turbid samples if necessary. Check the pH of the sample
and, if necessary, adjust to pH 7.0 with sulfuric acid or sodium
hydroxide solution.

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1.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2516 resp. 2517 or 2518 „ADMI”.

Zeroing the photometer:

•
• The photometer must be zeroed prior to each new measurement series.
• It is advisable to zero the photometer using the same cell as the one used for the measurement
sample or else a cell with identical optical characteristics and an identical absorption (matched pair).
The zero-adjustment procedure for the measurement series is automatically prompted by the instru-
ment.

• For the zero adjustment fill a corresponding rectangular cell with water for analysis.
• After prompting, insert the filled rectangular cell into the cell compartment. The zero adjustment is
performed automatically.
• Confirm the performance of the zero-adjustment procedure by clicking on <OK> 1 .
The zero adjustment is valid for the entire measurement series.

• Measurement:
Note
It is advisable to measure the sample using the same cell as the one used for the zero adjustment or else
a cell with identical optical characteristics and an identical absorption (matched pair).

• Fill the sample solution into a corresponding rectangular cell and insert the cell into the cell compart-
ment. The measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

The measurement result appears in the photometer display 4 .

• Click <START> 5 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

1.7 Evaluation

Statement of the results: ADMI Units

1.8 Method control

The method can be checked using Cat. No. 1.00246.0250 – Platinum cobalt color reference solution (HAZEN
500). Dilute this solution to a Hazen value in the middle of the measuring range with water for analysis or
distilled water and analyze.

1.9 Adjustment (Method 2517 and 2518 only)

• The ADMI factor of 1365 used in the calculation of the measurement result can be adjusted by the user.
The corrected ADMI factor must then be recalculated as follows:

ADMI factor corrected = ADMI factor x (specified method-check value / measured method-check value)

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• To alther the ADMI factor, select Method 2517 or Method 2518 from <Methods>.

• Click <Settings> 6 and select the list “FACTORS” 7 .

Adjustment is not possible in Method 2516, since this method uses a nonlinear function in the last stage of
the calculation of the ADMI factor to ensure an extended measuring range up to ADMI factors of 1000.

• Tip on the input field “ Factor” 8 , enter the corrected ADMI factor, and confirm by clicking on
<OK> 9 .

1.10 Literature

1. A
llen, W.; Prescott, W. B.; Derby, R. E.; Garland, C. E.; Peret, J. M.; Saltzman, M.; 1973. Determination
of color of water and wastewater by means of ADMI color values. Proc. 28th Ind. Waste Conf., Purdue
Univ., Eng. Ext. Ser. No. 142:661ff
2. S
tandard Methods for the Examination of Water and Wastewater (21th Edition); APHA Method 2120
Color -F. ADMI Weighted-Ordinate Spectrophotometric Method
3. M
cClaren; The Adams-Nickerson-Color Difference Formula; J. of the Society of Dyers and Columnists;
86, 354ff
4. Bridgeman; Inversion of the Munsell value equoation; J. of Optical Society of America; 1953; 499ff

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2 Anisidine value
2.1 Method

The Anisidine value is a measure for the amount of α, β-unsaturated aldehydes (2-alkenals) in animal and
vegetable fats and oils. It is defined as the 100-fold volume of the absorbance of a reaction solution mea-
sured at 350 nm in a 10-mm rectangular cell, relative to a concentration of the sample solution of 0.01 g/ml.
In the analysis, the sample under investigation is dissolved in isooctane, followed by the addition of p-anisi-
dine. After being left to react for 10 minutes, the absorbance is measured at 350 nm and the Anisidine
Value is calculated.
The method is analogous to the ISO 6885 standard (see literature reference1) and to DGF C-VI 6e (see liter-
ature reference2).

2.2 Measuring range

Method 2587 Anisidine value 0.0 - 200.0 AV (Anisidine value)

(10-mm rectangular cell)

2.3 Sample material

Animal and vegetable fats and oils

2.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114946 - Rectangular cells 10 mm
• Cat. No. 104718 - Isooctane Uvasol®
• Cat. No. A88255 - p-Anisidine (Caution! Carcinogenic!)
• Cat. No. 106649 - Sodium sulfate anhydrous for analysis EMSURE®
• Cat. No. 100063 - Acetic acid (lacial) 100% anhydrous for analysis EMSURE®
• Cat. No. 106652 - Sodium sulfite anhydrous EMPROVE® (optional)
• Cat. No. 102186 - Charcoal activated for analysis (optional)
• Paper filters, medium pore size (optional)
• 25-ml volumetric flasks
• 50-ml volumetric flasks
• Heating bath or drying cabinet (optional)
Standard laboratory glassware (e. g. glass beakers) and pipettes

2.5 Preparing the solutions

p-Anisidine, anhydrous cream-colored crystals:

•
The p-anisidine must not show any color (gray or pink). If it is colored, the p-anisidine must be purified
according to the ISO 6885 (see literature reference1) or the DGF C-VI 6e method (see literature refer-
ence2).

• Anisidine reagent:
The reagent must be prepared freshly on each new working day according to the ISO 6885 (see literature
reference1) or the DGF C-VI 6e method (see literature reference2).

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2.6 Preparation

• I f the moisture content of the sample is higher than w = 0.10%, the sample must be dried with sodium
sulfate according to the ISO 6885 (see literature reference1) or the DGF C-VI 6e method (see literature
reference2).
• W
eigh in a sufficient quantity of the sample (generally 0.4 g - 4.0 g), accurately weighed to 1 mg, into a
25-ml volumetric flask, note the sample weight, dissolve in 5 - 10 ml of isooctane, and subsequently
make up to the mark with isooctane. Solid samples can be heated up to a temperature 10 °C above their
melting point before they are weighed.
The instructions given in the ISO 6885 (see literature reference1) or the DGF C-VI 6e method (see litera-
ture reference2) must be followed.

2.7 Procedure and measurement

• Pre-reaction test solution (A0)

• Place 5 ml of the pretreated sample in a closeable vessel (e.g. a conical flask with a ground-glass stop-
per) and add 1 ml of acetic acid. Close the vessel and shake vigorously.
Leave the mixture to stand at room temperature in a dark place for 8 minutes.
Post-reaction test solution (A1)
•
• Place 5 ml of the pretreated sample in a closeable vessel (e.g. a conical flask with a ground-glass stop-
per) and add 1 ml of ansidine reagent acid. Close the vessel and shake vigorously.
Leave the mixture to stand at room temperature in a dark place for 8 minutes.
• Blank solution (A2)
• Place 5 ml of sooctane in a closeable vessel (e.g. a conical flask with a ground-glass stopper) and add
1 ml of ansidine reagent acid. Close the vessel and shake vigorously.
Leave the mixture to stand at room temperature in a dark place for 8 minutes.

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions and the blank solution or else a cell with identical optical characteristics and
an identical absorption (matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 2587 „Anisidine Value”.

2 1

A window with an input field to enter the sample weight pops up. Close the input field without entering a
sample weight by clicking on <X> 1 and click on the button <Settings> 2 .

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After clicking on the button <Settings> a window with selection options pops up.
• Select “ZERO ADJUSTMENT” 3 . The window changes.

• For the zero adjustment fill a clean and dry 10-mm rectangular cell with isooctane.
• After prompting, insert the filled rectangular cell into the cell compartment. The zero adjustment is
performed automatically.
• Confirm the performance of the zero-adjustment procedure by clicking on <OK> 4 .

• Measurement:
Note
It is advisable to measure the test solutions and the blank solution using the same cell as the one used
for the zero adjustment or else a cell with identical optical characteristics and an identical absorption
(matched pair).

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• Open the methods list (<Methods>) and select Method No. 2587 „Anisidine Value”. A window
with an input field to enter the sample weight pops up.

• Enter the weight of the sample in grams (g), accurate to 0.001 grams (g) 5 , confirm with <OK> 6 ,
and click on <START> 7 to switch to the measurement procedure.

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• Fill the post-reaction test solution (A1) into a clean and dry 10-mm rectangular cell after the reaction
time of 8 minutes; after the entire reaction time of 10 ± 1 minutes insert the cell into the cell compart-
ment. The measurement is performed automatically. A () symbol appears behind the cue “Insert
eacted solution (A1)” 8 .
• Confirm the measurement by clicking on <OK> 9 .

• Subsequently fill the pre-reaction test solution (A0) into a clean and dry 10-mm rectangular cell after
the reaction time of 8 minutes; after the entire reaction time of 10 ± 1 minutes insert the cell into the
cell compartment. The measurement is performed automatically. A () appears behind the cue “Insert
unreacted solution (A0)” 10 .
• Confirm the measurement by clicking on <OK>.

• Finally fill the blank solution (A2) into a clean and dry 10-mm rectangular cell after the reaction time of
8 minutes; after the entire reaction time of 10 ± 1 minutes insert the cell into the cell compartment.
The measurement is performed automatically. A () appears behind the cue “Insert blank solution
(A2)” 11 .
• Confirm the measurement by clicking on <OK>.

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14
13

The measurement result for the Anisidine value and the absorption values A1, A0, A2 appear in the pho-
tometer display 12 .
• Where applicable, use the scroll keys 13 to scroll down through the display to show other measure-
ment results.
• Click <START> 14 to start the measurement procedure for the next sample.

2.8 Evaluation

Statement of the results: Anisidine value

Absorptions of test solutions A1, A0, A2

2.9 Literature

1. ISO 6885:2016 Animal and vegetable fats and oils - Determination of anisidine value
2. DGF-Einheitsmethoden (2. Auflage einschließlich 14. Akt.-Lfg von 2009) - Abteilung Fette - C-VI 6e -
Anisidinzahl

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3 ASTM color measurement

3.1 Method

The ASTM color scale is used to evaluate the color of liquid petroleum products with a color darker than that
of the Saybolt color scale. The scale spans the range from 0.5 (lightest color) to 8.0 (darkest color).
The value is determined by measuring the transmittances in the wavelength spectrum between 380 - 780
nm and subsequent calculation of the tristimulus values X, Y, Z. In further calculation steps according to the
ASTM D6045 standard (see literature reference1) the tristimulus values are taken as a basis for the calcula-
tion of the ASTM color value.
The method is analogous to the ASTM D6045 procedure (see literature reference1).

3.2 Measuring range

Method 2562 ASTM Color 0.5 - 8.0 ASTM

(10-mm rectangular cell)

3.3 Sample material

Liquid samples
Petroleum products (e. g. uncolored gasoline and aviation fuel, naphtha, kerosine, pharmaceutical white
mineral oil, diesel oils, fuel oils, lubricant oils)

3.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114946 - Rectangular cells 10 mm
• Cat. No. 116754 - W
ater for analysis EMSURE® or
distilled water
• Ultrasound bath (optional)
• Heating bath (optional)
• Filter paper or centrifuge (optional)

3.5 Preparation

• Eliminate any air bubbles present in the sample by degassing the sample in the ultrasound bath.
• Centrifuge turbid samples and use the supernatant or else filter over a paper filter and use the filtrate.
• Melt solid or waxlike samples in a water bath with gentle heating and subsequently homogenize by
stirring. Take care not to overheat the sample in the melting process.

3.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2562 „ASTM Color”.

Zeroing the photometer:

•
• The photometer must be zeroed prior to each new measurement series.
• It is advisable to zero the photometer using the same cell as the one used for the measurement sam-
ple or else a cell with identical optical characteristics and an identical absorption (matched pair).
The zero-adjustment procedure for the measurement series is automatically prompted by the instru-
ment.

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• For the zero adjustment fill a 10-mm rectangular cell with water for analysis.
• After prompting, insert the filled rectangular cell into the cell compartment. The zero adjustment is
performed automatically.
• Confirm the performance of the zero-adjustment procedure by clicking on <OK> 1 .
The zero adjustment is valid for the entire measurement series.

• Fill the sample solution into a 10-mm rectangular cell and insert the cell into the cell compartment.
The measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

The measurement result appears in the photometer display 4 .

• Click <START> 5 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

3.7 Evaluation

Statement of the results: ASTM Units

3.8 Literature

1. ASTM D6045-12, Standard Test Method for Color of Petroleum Products by the Automatic Tristimulus
Method

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4 CIE color distance

4.1 Method

Differences in color can be quantified by measuring the color distance ΔE*ab (also referred to as Delta E).
The coordinates of the colorimetric loci (L*a*b*) of the samples to be compared are used to determine their
positions in the color space (colorimetric loci). Subsequently the Euclidean distance between the colorimet-
ric loci is calculated and the result is defined as the color distance (ΔE value). In the CIELAB color space
(see section 5, “Chroma (CIELAB color space)”), the values for the brightness difference (ΔL*), the distance
of the coordinates a*, b* (Δa* and Δb*), and the chromatic difference (ΔC*ab) also play a relevant role.
These values are determined by the spectrophotometric measurement of a reference sample and an analy-
sis sample. The transmittances for both samples in the wavelength spectrum between 360 and 780 nm are
measured, and subsequently the color distance, the brightness difference, the distance of the coordinates
a* and b*, and the chromatic difference are determined according to the methods of the DIN EN ISO
11664-4 standard (see literature reference 1).
This method is based on the DIN EN 11664-4 (see literature reference1) and DIN 5033-3 standards (see lit-
erature reference2).

4.2 Measuring range

Method 2584 CIE Color Distance D65/2° ΔE*ab 0.00 - 200.00

ΔL* -200.00 - 200.00
Δa*-200.00 - 200.00
Δb*-200.00 - 200.00
ΔC*ab -200.00 - 200.00
(10-mm rectangular cell)

Note
The measurement values are determined for standard illuminant D65 and the 2° standard observer.

4.3 Sample material

Clear liquid samples

Turbid liquid samples after filtration

4.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

4.5 Preparation

• Filter turbid sample solutions over a membrane filter.

4.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2584 „CIE Color Distance D65/2°”.

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Zeroing the photometer:

• Measurement:
Note
It is advisable to measure the reference solution and the measurement sample using the same cell as the
one used for the zero adjustment or else a cell with identical optical characteristics and an identical
absorption (matched pair).

• Fill the reference solution into a 10-mm rectangular cell and insert the cell into the cell compartment.
The measurement is performed automatically.

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A () appears behind the cue “Insert reference (R)” 2 .

• Confirm the measurement by clicking on <OK> 3 .
• Fill the sample solution into a 10-mm rectangular cell and insert the cell into the cell compartment. The
measurement is performed automatically.

A () appears behind the cue “Insert sample (T)” 4 .

• Confirm the measurement by clicking on <OK> 5 .

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7
6

The measurement results appear in the photometer display 5 .

• Where applicable, use the scroll keys 6 to scroll down through the display to show other measure-
ment results.
• Click <START> 7 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

4.7 Evaluation

E*ab value
Statement of the results: Δ
ΔL* value
Δa* value
Δb* value
ΔC*ab value
L*a*b* values for the reference sample are shown with a lower-case “R” (e. g. L*R)
L*a*b* values for the analysis sample are shown with a lower-case “T” (e. g. L*T)

C*ab,R value for the reference sample

C*ab,T value for the analysis sample
determined for standard illuminant D65 and the 2° standard observer

4.8 Literature

1. DIN EN ISO 11664-4:2012-06, Colorimetry - Part 4: CIE 1976 L*a*b* Colour space
2. DIN 5033-3:1992-07, Colorimetry; Colorimetric measures
3. ASTM E-308-06, Standard Practice for Computing the Colors of Objects by Using the CIE System

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5 CIELAB color space (brightness, chroma)

5.1 Method

The CIELAB color space, also known as CIE-L*a*b* or simply Lab, describes all perceivable colors and is the
most widely used color space. The CIELAB color space is based on the XYZ model set up by the CIE organi-
zation in 1931 (CIE – Commission internationale de l’éclairage, International Commission on Illumination).
This model uses the spectral characteristics of a color sample to calculate the so-called tristimulus values
X, Y, Z, which stand in direct correlation to the intensity of the stimulation of the three types of color recep-
tors (cone cells) of the human eye (red, green, blue). Since the XYZ model proved to be difficult to handle
in the assessment of color differences, in 1976 the CIE published the CIELAB model. The CIELAB model is
better capable of describing the human perception of color than the previous XYZ model, since color-per-
ception differences are presented in Euclidean distances within a Cartesian coordinate system. In this coor-
dinate system, each color can be defined by its coordinates L*, a*, and b*.
On the a* axis, green and red lie opposite each other, on the b* axis blue and yellow. Gray lies at the coor-
dinate origin (a*=0 and b*=0). The L* axis stands in the coordinate origin vertically at the a*b* level and
describes the brightness of the color, with values ranging between 0 and 100.
Negative a* values stand for green colors, positive a* values for red colors. On the b* axis, negative b* val-
ues describe blue colors, with positive b* values describing yellow colors.
In practice, the C*ab value – the chroma – is also frequently calculated. The chroma expresses the relative
color effect of a sample in relation to a reference white.
The L*a*b* values and the chroma C*ab value are determined by measuring the transmittances in a wave-
length spectrum between 360 and 780 nm and subsequently calculating the L*a*b* and chroma C*ab val-
ues according to the methods of the DIN EN ISO 11664-4 standard (see literature reference1).
The method is based on the DIN EN 11664-4 (see literature reference1) and DIN 5033-3 standards (see lit-
erature reference2).

5.2 Measuring range

Method 2580 CIELAB D65/2° L* 0.00 - 105.00

a* -180.0 - 180.0
b* -180.0 - 180.0
C*ab 0.00 - 300.00
(10-mm rectangular cell)

Note
The measurement values are determined for standard illuminant D65 and the 2° standard observer.

5.3 Sample material

Clear liquid samples

Turbid liquid samples after filtration

5.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

5.5 Preparation

• Filter turbid sample solutions over a membrane filter.

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5.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2580 “CIELAB D65/2°”.

Zeroing the photometer:

• Fill the sample solution into a 10-mm rectangular cell and insert the cell into the cell compartment. The
measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

6
5

The measurement results appear in the photometer display 4 .

• Where applicable, use the scroll keys 5 to scroll down through the display to show other measure-
ment results.
• Click <START> 6 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

5.7 Evaluation

Statement of the results: L

*a*b* values
C*ab value
Tristimulus values X, Y, Z
determined for standard illuminant D65 and the 2° standard observer

5.8 Literature

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6 CIELUV color space

6.1 Method

The CIELUV color room, also referred to as CIE-L*u*v*, was developed in 1976 by the CIE organization and
– similar to the CIELAB color space – describes all perceivable colors. Like the CIELAB color space, the
CIELUV color room is based on the CIE’s XYZ model from 1931. This model takes the spectral characteristics
of a color sample to calculate the so-called tristimulus values X, Y, Z, which stand in direct correlation to the
intensity of the stimulation of the three types of color receptors (cone cells) of the human eye (red, green,
blue). The CIELUV color room is predominantly used for the measurement of light colors of luminescent
objects and of the additive color mixture. Compared with the CIELab color room, the CIELUV color room
possesses a reduced green spectrum and an enlarged blue spectrum.
In this coordinate system, each color can be defined by its coordinates L*, u*, and v*.
On the u* axis, green and red lie opposite each other, on the v* axis blue and yellow. Gray lies at the coor-
dinate origin (u*=0 and v*=0). The L* axis stands in the coordinate origin vertically at the u*v* level and
describes the brightness of the color, with values ranging between 0 and 100.
Negative u* values stand for green colors, positive u* values for red colors. On the v* axis, negative v* val-
ues describe blue colors, with positive v* values describing yellow colors.
In practice, the C*uv value – the chroma – as well as the S*uv value – the saturation – are also frequently
calculated. The chroma expresses the relative color effect of a sample in relation to a reference white (white
point). The saturation describes the distance of the colorimetric locus from the achromatic axis, on which lie
all achromatic colors between white and black.
The L*u*v* values, the the chroma C*uv value, and the saturation S*uv value are determined by measur-
ing the transmittances in a wavelength spectrum between 360 and 780 nm and subsequently calculating the
L*u*v, the chroma C*uv, and the S*uv values according to the methods of the DIN 5033-3 (see literature
reference1) and ISO CIE 11664-5 standards (see literature reference2).
The method is based on the DIN 5033-3 (see literature reference 1) and ISO CIE 11664-5 standards (see
literature reference2).

6.2 Measuring range

Method 2581 CIELUV D65/2° L* 0.00 - 105.00

u* -180.0 - 180.0
v* -180.0 - 180.0
C*uv 0.00 - 300.00
S*uv 0.000 - 200.000
(10-mm rectangular cell)

Note
The measurement values are determined for standard illuminant D65 and the 2° standard observer.

6.3 Sample material

Clear liquid samples

Turbid liquid samples after filtration

6.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

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6.5 Preparation

• Filter turbid sample solutions over a membrane filter.

6.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2581 “CIELUV D65/2°”.

• Zeroing the photometer:

• The photometer must be zeroed prior to each new measurement series.
• It is advisable to zero the photometer using the same cell as the one used for the measurement sam-
ple or else a cell with identical optical characteristics and an identical absorption (matched pair).
The zero-adjustment procedure for the measurement series is automatically prompted by the instru-
ment.

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• Fill the sample solution into a 10-mm rectangular cell and insert the cell into the cell compartment. The
measurement is performed automatically.

A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

6
5

The measurement results appear in the photometer display 4 .

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6.7 Evaluation

Statement of the results: L

*u*v* values
C*uv value
S*uv value
Tristimulus values X, Y, Z
determined for standard illuminant D65 and the 2° standard observer

6.8 Literature

1. DIN 5033-3:1992-07, Colorimetry; Colorimetric measures

2. ISO CIE 11664-5:2016(E), Colorimetry - Part 5: CIE 1976 L*u*v* colour space and u’, v’ uniform
chromaticity scale diagram
3. ASTM E-308-06, Standard Practice for Computing the Colors of Objects by Using the CIE System

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7 CIExyY color space

7.1 Method

The CIExyY color space is based on the CIE Chromacity Diagram from the year 1931 and is one of the old-
est known color spaces. The CIExyY color space and the chromaticity diagram are derived from the CIE XYZ
model. This model takes the spectral characteristics of a color sample to calculate the so-called tristimulus
values X, Y, Z, which stand in direct correlation to the intensity of the stimulation of the three types of color
receptors (cone cells) of the human eye (red, green, blue). The XYZ model exhibits a number of weakpoints
when it comes to the assessment of color differences. The CIE subsequently developed a two-dimensional
graph with x/y coordinates derived from the tristimulus values X, Y, Z. This produced a graphic model (the
CIE Chromacity Diagram) with its characteristic horseshoe or shoesole shape, which can be used to define
the location of a color or color space. In the CIExyY color space the value Y is also added, which corre-
sponds to the brightness or the relative luminance of a color.
In the CIExyY color space x values stand for the red/purple axis and y values for the green axis. The values
of the Y axis can be depicted only in the three-dimensional diagram.
The CIExyY values are determined by measuring the transmittances in a wavelength spectrum between 360
and 780 nm and subsequently calculating the xyY values according to the methods of CIE 15:2004, Techni-
cal Report - Colorimetry (see literature reference1).

7.2 Measuring range

Method 2582 CIExyY D65/2° x 0.0000 - 0.8000

y 0.0000 - 0.8000
Y 0.000 - 200.000
(10-mm rectangular cell)

Note
The measurement values are determined for standard illuminant D65 and the 2° standard observer.

7.3 Sample material

Clear liquid samples

Turbid liquid samples after filtration

7.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

7.5 Preparation

• Filter turbid sample solutions over a membrane filter.

7.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2582 “CIExyY D65/2°”.

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Zeroing the photometer:

• Fill the sample solution into a 10-mm rectangular cell and insert the cell into the cell compartment. The
measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

The measurement results appear in the photometer display 4 .

• Click <START> 5 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

7.7 Evaluation

Statement of the results: x

yY values
determined for standard illuminant D65 and the 2° standard observer

7.8 Literature

1. CIE 15:2004, Technical Report - Colorimetry, ISBN 978-3-901906-33-6

2. ASTM E-308-06, Standard Practice for Computing the Colors of Objects by Using the CIE System

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8 Color (ASBC method)

8.1 Method

The spectrophotometric method is an official method of the ASBC (American Society of Brewing Chemists)
for determining the color of beers, worts, and liquid malt substitutes.
The color is determined by measuring the absorption of the sample at 430 nm and subsequently converting
the result into °SRM using a factor specified in the literature (see literature references1, 2). Taking an addi-
tional measurement of the absorption at 700 nm enables the assessment of any potential turbidity of the
sample.
The method is analogous to the ASBC Method Beer-10 (see literature reference1) and ASBC Method Wort-9
(see literature reference2).

8.2 Measuring range

Method 2633 Color - ASBC 0.0 - 50.0 °SRM

(10-mm rectangular cell)

0.0 - 100.0 EBC Units

(10-mm rectangular cell)

8.3 Sample material

Beers, production worts, laboratory worts, liquid malt substitutes

8.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114946 - Rectangular cells 10 mm
ater for analysis EMSURE® or
• Cat. No. 116754 - W
distilled water
• Cat. No. 107910 - Kieselguhr GR for analysis (optional)
• Membrane filters, pore size max. 0.45 µm (optional)
• Paper filters (optional)

8.5 Preparation

• Degas the sample to remove any carbon dioxide.

• Filter the sample over a membrane filter; filtration can be dispensed with in the case that the ratio of the
absorption values from 700 nm to 430 nm is ≤ 0.039 (A700 nm / A430 nm) (the absorption values at 700 nm
and 430 nm and the quotient “A700 nm / A430 nm” are also shown together with the result after the measure-
ment).
• If necessary, clarify the sample by adding 0.1% kieselguhr (Kieselguhr GR, Cat. No. 107910) and filtra-
tion before the membrane-filtration step
• In the case of a result °SRM Units > 50.0, dilute the sample in such a way that its color is within the
measurement range, noting the dilution factor.
• Worts must be filtered at 5 - 8°C directly after taking the sample or otherwise preserved prior to filtra-
tion (by pasteurization or storage in a refrigerator). After filtration, add 5 g of kieselguhr to 100 ml of fil-
tered wort, swirl to mix thoroughly, and leave to stand for 5 min. Subsequently filter the suspension over
a paper filter. Refilter the first 30 - 40 ml and transfer the collected filtrate in a clean vessel until required
for analysis.

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8.6 Procedure and measurement

Zeroing the photometer:

1
2

A window with an input field to enter the dilution factor pops up. Close the input field without entering
a ilution factor by clicking on <X> 1 and click on the button <Settings> 2 .

After clicking on the button <Settings> a window with selection options pops up.
• Select “ZERO ADJUSTMENT” 3 . The window changes.

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• Measurement:
Note
It is advisable to measure the sample solution using the same cell as the one used for the zero adjust-
ment or else a cell with identical optical characteristics and an identical absorption (matched pair).
• Open the methods list (<Methods>) and select Method No. 2633 “Color - ASBC”. A window with
an input field to enter the dilution factor pops up.

• Samples with a value > 50.0 °SRM must be diluted with water.
Enter the dilution factor in the proportion of 1 part sample + x parts water 5 and confirm by clicking
on <OK> 6 (e. g. 1 + 9, when 10 ml of sample is mixed with 90 ml of water).
If an undiluted sample is being measured, enter the value “0” and confirm with<OK> 6 .

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• Click on <START> 7 to switch to the measurement procedure.

• Fill the sample solution into a 10-mm rectangular cell and insert the cell into the cell compartment. The
measurement is performed automatically.

A () appears behind the cue “Insert cell” 8 .

• Confirm the measurement by clicking on <OK> 9 .

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12
11

The measurement results in the units °SRM and EBC as well as the absorptions at 430 nm and 700 nm
and also the quotient “A700 nm / A430 nm” appear in the photometer display 10 .
• Where applicable, use the scroll keys 11 to scroll down through the display to show other measure-
ment results.
• Click <START> 12 to start the measurement procedure for the next sample.

8.7 Evaluation

Statement of the results: °

SRM
EBC Units
Absorption at 430 nm and 700 nm
Quotient of the absorptions at 700 nm and 430 nm (A700 nm / A430 nm)

Note
If the quotient “A700 nm / A430 nm” is at a value ≥ 0.039, the sample must be filtered (see section 8.5,
“Preparation”).

Note
A spectrophotometric absorption curve does not reflect the color impression of the human eye, since light of
the same intensity in different parts of the spectrum has different effects on the eye. In addition, the
absorption curves at 430 nm are very steep, meaning that measurement errors can easily happen. What is
more, there are differences that occur in the comparison of light beers with diluted dark beers.

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8.8 Literature

1. ASBC Methods of Analysis , online. Beer-10, Color, A. Spectrophotometric color method [Release date
1958, revised 1975, reviewed 2015].
American society of brewing Chemists, St. Paul, Mn, U.S.A. doi: 10.1094/ASBCMOA-Beer-10
2. ASBC Methods of Analysis , online. Wort-9, Wort Color and Sample Preparation, A. Celite [Release date
1969, revised 1976, reviewed 2010].
American society of brewing Chemists, St. Paul, Mn, U.S.A. doi: 10.1094/ASBCMOA-Wort-9

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9 Color (EBC method)

9.1 Method

This spectrophotometric method counts as the official method of the EBC (European Brewery Convention)
for determining the color of beers, worts, and liquid malt substitutes.
The color is determined by measuring the absorption of the sample at 430 nm and subsequently converting
the result into EBC units using a factor specified in the literature (see literature references1, 2, 3).
This method is analogous to the MEBAK Method 2.13.2 (see literature reference1), EBC Method 8.5 (see lit-
erature reference2), and EBC Method 9.6 (see literature reference3).

9.2 Measuring range

Method 2602 Color - EBC 0.0 - 60.0 EBC Units

(10-mm rectangular cell)

9.3 Sample material

Beers, production worts, laboratory worts, liquid malt substitutes

9.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114946 - Rectangular cells 10 mm
• Cat. No. 116754 - W
ater for analysis EMSURE® or
distilled water
• Cat. No. 107910 - Kieselguhr GR for analysis (optional)
• Membrane filters, pore size max. 0.45 µm (optional)

9.5 Preparation

• Degas the sample to remove any carbon dioxide.

• Filter the sample over a membrane filter; filtration can be dispensed with in the case that the turbidity of
the diluted sample is below 1 EBC turbidity units.
• If necessary, clarify the sample by adding 0.1% kieselguhr (Kieselguhr GR, Cat. No. 107910) and filtra-
tion before the membrane-filtration step
• In the case of a result EBC Units > 60.0, dilute the sample in such a way that its color is within the mea-
surement range; include the dilution factor when calculating the final result (see Note “Dilution”, section
9.6).

9.6 Procedure and measurement

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 2602 „Color - EBC”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

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If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• Open the methods list (<Methods>) 4 and select Method No. 2602 „Color - EBC”.

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• Fill the sample solution into a 10-mm rectangular cell and insert the cell into the cell compartment. The
measurement is performed automatically and the result appears in the display 5 .

Note „Dilution“

If the sample exhibits a color stronger than 60.0 EBC, it must be diluted with water for analysis until the
color is within the measurement range. The dilution factor must be taken into account in the calculation
of the result and can be entered into the photometer.

Do this by clicking on the button <Settings> 6 and subsequently selecting the field “DILUTION” 7 .

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A window with an input field to enter the dilution factor is opened.
Activate the input field behind the “+” character 8 and enter the dilution factor in the ratio of 1 part
sample + x parts water and confirm by clicking on <OK> 9 (e. g. 1 + 9, when 10 ml of sample is mixed
with 90 ml of water).

The window changes to the methods display. The activated dilution factor appears in the display 10 and is
automatically taken into account in the calculation of the result 11 .

9.7 Evaluation

Statement of the results: E

BC Units

9.8 Literature

1. MEBAK Brautechnische Analysemethoden 4. Auflage 2002, Band II, Method 2.13.2, Seite 88ff
2. Analytica-EBC, section 8 Wort, Method 8.5 (2000-10)
3. Analytica-EBC, section 9 Beer, Method 9.6 (2000-10)

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10 Color Hazen
10.1 Method

The color according to Hazen, also known as the Hazen color index, APHA color, or Pt/Co color index, is used
to assess the color of clear liquids that possess color characteristics similar to those shown on the platinum/
cobalt scale.
This method was developed by Allen Hazen in 1892 and was originally based on a visual comparison of the
color of the sample solution with a defined color standard prepared from platinum and cobalt salts in acidic
solution.
The method is described in various regulations and is referenced in many technical textbooks. The originally
visual color comparison has meanwhile been replaced by photometric methods, since these enable a far
higher degree of accuracy to be achieved.
The regulations and technical textbooks describe a broad variety of measurement conditions. An aspect
common to all sources of reference, however, is that the measurement results achieved under the respec-
tive measurement conditions must be referenced and traceable to a defined color standard consisting of
platinum and cobalt salts in acidic solution.
The “Color Hazen” method can be run on the Spectroquant® Prove spectrophotometers at various wave-
lengths:
• 465 nm (analogous to DIN EN ISO 6271 - see literature reference1, analogous to APHA 2120C - see liter-
ature reference2, analogous to ASTM D 1209-05 - see literature reference3)
• 455 nm (analogous to DIN EN ISO 6271 - see literature reference1, analogous to APHA 2120C - see liter-
ature reference2, analogous to ASTM D 1209-05 - see literature reference3)
• 445 nm (analogous to DIN EN ISO 6271 - see literature reference1, analogous to ASTM D 1209-05 - see
literature reference3)
• 340 nm
The measurement result can be expressed in CU (color units) or Hazen units, with the unit being is defined
as mg of platinum per litre of solution. The regulations and technical textbooks also use units such as mg/l
Pt/Co and mg/l Pt. The conversion factor between the units is 1.

10.2 Measuring range

Method 32 Color Hazen 340 0.2 - 100.0 CU

(50-mm rectangular cell)

1 - 250 CU
(20-mm rectangular cell)

1 - 500 CU
(10-mm rectangular cell)

Method 179 Color Hazen 445 1 - 1000 CU

(50-mm rectangular cell)

Method 180 Color Hazen 455 1 - 1000 CU

(50-mm rectangular cell)

Method 181 Color Hazen 465 1 - 1000 CU

(50-mm rectangular cell)

10.3 Sample material

Clear liquid samples with color characteristics similar to those on the Pt/Co color scale (= yellow to
yellow-brown)
Turbid liquid samples with color characteristics after filtration similar to those on the Pt/Co color scale
(= yellow to yellow-brown)

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10.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

10.5 Preparation

• Filter turbid sample solutions over a membrane filter.

10.6 Procedure and measurement

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 32 “Color Hazen 340” resp. No. 179 “Color Hazen 445” or No. 180 “Color Hazen 455” or
No. 181 “Color Hazen 465”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

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If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

• For the zero adjustment fill a suitable rectangular cell with water for analysis.
• After prompting, insert the filled rectangular cell into the cell compartment. The zero adjustment is
performed automatically.
• Confirm the performance of the zero-adjustment procedure by clicking on <OK> 3 .

Measurement:
•
Note
It is advisable to measure the sample solution using the same cell as the one used for the zero adjust-
ment or else a cell with identical optical characteristics and an identical absorption (matched pair).

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• Open the methods list (<Methods>) 4 and select Method No. 32 “Color Hazen 340” resp.
No. 179 “Color Hazen 445” or No. 180 “Color Hazen 455” or No. 181 “Color Hazen 465”.

• Fill the sample solution into a corresponding rectangular cell and insert the cell into the cell compart-
ment. The measurement is performed automatically and the result appears in the display 5 .

10.7 Evaluation

Statement of the results: C

U or
Hazen or
mg/l Pt/Co or
mg/l Pt

10.8 Method control

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10.9 Recalibration

The methods can be recalibrated using Cat. No. 1.00246.0250 – Platinum cobalt color reference solution
(HAZEN 500). Recalibration can be done by measuring value pairs, by entering value pairs, or by entering
coefficients of a calibration function. Please refer to the Operating Manual of your spectrophotometer for
details on the recalibration procedure.

10.10 Literature

1. DIN EN ISO 6271 - Clear liquids - Estimation of colour by the platinum-cobalt colour scale
(ISO 6271:2015); German version EN ISO 6271:2015
2. Standard Methods for the Examination of Water and Wastewater (21th Edition), APHA 2120 Color - C.
Spectrophotometric - Single-Wavelength Method
3. ASTM D 1209-00 - Standard Test Method for Color of Clear Liquids (Platinum-Cobalt-Scale)
4. Water Research Vol. 30, No.11, 2771-2775, 1996 - Hongve and Akesson, Spectrophotometric
Determination of Water Colour in Hazen Units

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11 Color 410 acc. to EN 7887

11.1 Method

The examination and determination of color 410 according to the EN 7887 standard serves the assessment
of water quality. This procedure determines the true color of a filtered water sample caused by dissolved
substances.
The measurement is made at a wavelength of 410 nm. This is the shortest wavelength at which the absorp-
tion spectrum of a natural water sample with a color factor of 100 mg/l Pt overlaps with that of a corre-
sponding color reference solution.
The method is referenced and traceable to a defined color standard prepared from platinum and cobalt salts
in acidic solution (also known as the Hazen standard).
According to DIN EN ISO 7887, the measurement result is expressed as mg/l Pt. The technical literature,
however, also uses the units mg/l Pt/Co and CU (Color Unit). The conversion factor between the units is 1.
The method is analogous to the standard DIN EN ISO 7887:2011 - Method C (see literature reference1).

11.2 Measuring range

Method 303 Color 410 EN7887 2 - 500 CU

(50-mm rectangular cell)

5 - 1250 CU
(20-mm rectangular cell)

10 - 2500 CU
(10-mm rectangular cell)

11.3 Sample material

Water from water-treatment plants (crude water, drinking water, weakly colored industrial wastewater)

11.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

11.5 Preparation

• Filter turbid sample solutions over a membrane filter.

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11.6 Procedure and measurement

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 303 „Color 410 EN7887”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• Open the methods list (<Methods>) 4 and select Method No. 303 “Color 410 EN7887”.

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11.7 Evaluation

Statement of the results: m

g/l Pt or
mg/l Pt/Co or
CU

11.8 Method control

The method can be checked using Cat. No. 1.00246.0250 – Platinum cobalt color reference solution (HAZEN
500). When using a 50-mm rectangular cell, this solution must be diluted to a value in the middle of the
measuring range with water for analysis or distilled water and analyzed.

11.9 Recalibration

11.10 Literature

1. DIN EN ISO 7887:2011 - Water quality - Examination and determination of colour (ISO 7887:2011); Ger-
man version EN ISO 7887:2011

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12 Color - Spectral absorption coefficient acc. to DIN EN ISO 7887

12.1 Method

The examination and determination of color is used in the assessment of water quality. This procedure
determines the true color of a filtered water sample caused by dissolved substances.
The maximum absorption of a sample is dependent on its color (e. g. yellow, red, or green). This is why the
spectral absorption coefficient should be determined according to the standard DIN EN ISO 7887 - Method B
at three different wavelengths (436 nm, 525 nm, 620 nm).
The measurement results for the wavelengths and the respective spectral absorption coefficients are
expressed in the unit m-1.
The methods are analogous to the standard DIN EN ISO 7887:2011 - Method B (see literature reference1).
Methods Nos. 15 “Color 436” and 302 “SAC α(436)” are identical and differ solely in the method designa-
tion. Furthermore, the two methods are both analogous to the procedures prescribed in the German “Trink-
wasserverordnung” (Drinking Water Ordinance) Annex 3, No. 7 (see literature reference2).

12.2 Measuring range

Method 15 Color 436 0.1 - 50.0 m-1

(mono-wave- (50-mm rectangular cell)
length method,
436 nm) 0.3 - 125.0 m-1
(20-mm rectangular cell)

1.0 - 250 m-1

(10-mm rectangular cell)

Method 302 SAC α(436) 0.1 - 50.0 m-1

(mono-wave- (50-mm rectangular cell)
length method,
436 nm) 0.3 - 125.0 m-1
(20-mm rectangular cell)

1 - 250 m-1
(10-mm rectangular cell)

Method 61 Color 525 0.1 - 50.0 m-1

(mono-wave- (50-mm rectangular cell)
length method,
525 nm) 0.3 - 125.0 m-1
(20-mm rectangular cell)

1.0 - 250 m-1

(10-mm rectangular cell)

Method 78 Color 620 0.1 - 50.0 m-1

(mono-wave- (50-mm rectangular cell)
length method,
620 nm) 0.3 - 125.0 m-1
(20-mm rectangular cell)

1.0 - 250 m-1

(10-mm rectangular cell)

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Method 2588 Color EN 7887 0.0 - 50.0 m-1

(multi-wave- (50-mm rectangular cell)
length method,
436 nm, 525 nm, 0.0 - 125.0 m-1
620 nm) (20-mm rectangular cell)

0.0 - 250.0 m-1

(10-mm rectangular cell)

Note
The colorimetric measurement can be performed with a multi-wavelength method or consecutively with
mono-wavelength methods.

12.3 Sample material

Water from water-treatment plants (crude water, drinking water, weakly colored industrial wastewater)

12.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

12.5 Preparation

• Filter turbid sample solutions over a membrane filter.

12.6 Procedure and measurement - Mono-wavelength methods

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 15 “Color 436” resp. 302 “SAC α(436)” or 61 “Color 525” or 78 “Color 620”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

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2
1

If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• Open the methods list (<Methods>) 4 and select Method No. 15 “Color 436” resp. 302
“SAC α(436)” or 61 “Color 525” or 78 “Color 620”.

12.7 Procedure and measurement - Multiwavelength method

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 2588 “Color EN 7887”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

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If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 6 and clicking on the
selection button “ZERO ADJUSTMENT” 7 .

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• Open the methods list (<Methods>) 9 and select Method No. 2588 “Color EN 7887”.
• Fill the sample solution into a corresponding rectangular cell and insert the cell into the cell compart-
ment. The measurement is performed automatically.

A () appears behind the cue “Insert cell” 10 .

• Confirm the measurement by clicking on <OK> 11 .

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The measurement results at the wavelengths 436 nm, 525 nm, and 620 nm appear in the photometer
display 12 .
• Click <START> 13 to start the measurement procedure for the next sample.

12.8 Evaluation

Statement of the results: m

-1

12.9 Literature

1. DIN EN ISO 7887:2011 - Water quality - Examination and determination of colour (ISO 7887:2011);
German version EN ISO 7887:2011
2. Deutsche Trinkwasserverordnung - TrinkwV Anlage 3, No. 7

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13 Gardner color measurement

13.1 Method

The Gardner Color Scale is used to assess the color of clear, yellow to yellow-brownish liquids. The scale
spans a spectrum ranging from 1.0 (lightest color) to 18.0 (darkest color).
The color index is determined by measuring the transmittances of the sample in a wavelength spectrum
between 360 and 780 nm and subsequently calculating the tristimulus values X, Y, Z and the chromaticity
coordinates x and y. These chromaticity coordinates x and y values are then used to calculate the Gardner
Color Index according to the methods of the DIN EN ISO 4630-2 (see literature reference1) and ASTM
D6166 standards (see literature reference2).
The method is analogous to the DIN EN ISO 4630-2 (see literature reference1) and ASTM D6166 standards
(see literature reference2).

13.2 Measuring range

Method 2561 Gardner Color 1.0 - 18.0 Gardner

(10-mm rectangular cell)

13.3 Sample material

Clear yellow to yellow-brown liquids

Products made of natural resins (e. g. tall oils, tall-oil fatty acids, paints, oils)
Surfactants

13.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114946 - Rectangular cells 10 mm
ater for analysis EMSURE® or
• Cat. No. 116754 - W
distilled water
• Ultrasound bath (optional)
• Heating bath (optional)
• Filter paper or centrifuge (optional)

13.5 Preparation

13.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2561 “Gardner Color”.

Zeroing the photometer:

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• Fill the sample solution into a 10-mm rectangular cell and insert the cell into the cell compartment. The
measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

The measurement result appears in the photometer display 4 .

• Click <START> 5 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

13.7 Evaluation

Statement of the results: Gardner Units

13.8 Literature

1. DIN EN ISO 4630-2:2016-05, Clear liquids - Estimation of colour by the Gardner colour scale - Part 2:
Spectrophotometric method
2. ASTM D6166-12, Standard Test Method for Color of Pine Chemicals and Related Products
3. ASTM D1544-04, Standard Test Method for Color of Transparent Liquids (Gardner Color Scale)

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14 Hess-Ives color scale

14.1 Method

The Hess-Ives color scale serves to assess the color of fat derivatives and is used in the cosmetics industry.
The color index is determined from the weighted color intensities that correspond to the red, green, and
blue color components of the sample.
The index is determined by measuring the absorption at the wavelengths 460 nm, 470 nm, 560 nm, and
640 nm and subsequent calculations according to the procedure of DGK Test Method F 050.2.
The method is analogous to DGK Test Method F 050.2 (see literature reference1).

14.2 Measuring range

Method 2586 Hess-Ives Color 0 - 400 H-I

(10-mm rectangular cell)

0.0 - 80.0 H-I

(50-mm rectangular cell)

14.3 Sample material

Fat derivatives, surfactants

14.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114946 - Rectangular cells 10 mm and/or
Cat. No. 114944 - Rectangular cells 50 mm
ater for analysis EMSURE® or
• Cat. No. 116754 - W
distilled water
• Ultrasound bath (optional)
• Heating bath (optional)
• Filter paper or centrifuge (optional)

14.5 Preparation

• Eliminate any air bubbles present in the sample by degassing the sample in the ultrasound bath.
Centrifuge turbid samples and use the supernatant or else filter over a paper filter and use the filtrate.
•
Melt solid or waxlike samples in a water bath with gentle heating and subsequently homogenize by
•
stirring. Take care not to overheat the sample in the melting process.

14.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. Methodnliste (<Methodn (Methods)>) öff-
nen and Method No. 2586 “Hess-Ives Color”.

• Zeroing the photometer:

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• Fill the sample solution into a corresponding rectangular cell and insert the cell into the cell compart-
ment. The measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

The measurement result appears in the photometer display 4 .

• Click <START> 5 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

14.7 Evaluation

Statement of the results: H

ess-Ives (H-I) Units

14.8 Literature

1. DGK PrüfMethod F 050.2 (Deutsche Gesellschaft für wissenschaftliche and angewandte Kosmetik e.V.)

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15 Hunter color distance

15.1 Method

The color differences can be quantified by means of the color distance ΔE (also referred to as Delta E). The
coordinates of the colorimetric loci (L*a*b*) of the samples to be compared are used to determine their
position in the color space. Subsequently the Euclidean distance between the colorimetric loci is calculated
and the result is defined as the color distance (ΔE value). In the Hunter color space (see section 16, Hunter-
Lab color space) the values for the brightness difference (ΔL*) and the distance of the coordinates a*, b*
(Δa* and Δb*) also play a relevant role.
These values are determined by the spectrophotometric measurement of a reference sample and an analy-
sis sample. The transmittances for both samples in the wavelength spectrum between 360 and 780 nm is
measured, and subsequently the color distance, the brightness difference, and the distance of the coordi-
nates a* and b* are determined according to the methods of HunterLab Application Note Vol. 8, No. 9,
06/08 (see literature reference1).

15.2 Measuring range

Method 2585 Hunter Color Distance D65/2° ΔE*H 0.00 - 200.00

ΔL* -200.00 - 200.00
Δa*-200.00 - 200.00
Δb*-200.00 - 200.00
(10-mm rectangular cell)

Note
The measurement values are determined for standard illuminant D65 and the 2° standard observer.

15.3 Sample material

Clear liquid samples

Turbid liquid samples after filtration

15.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

15.5 Preparation

• Filter turbid sample solutions over a membrane filter.

15.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2585 “Hunter Color Distance D65/2°”.

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• Zeroing the photometer:

• Fill the reference solution into a 10-mm rectangular cell and insert the cell into the cell compartment.
The measurement is performed automatically.

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A () appears behind the cue “Insert reference (R)” 2 .

A () appears behind the cue “Insert sample (T)” 4 .

• Confirm the measurement by clicking on <OK> 5 .

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8
7

The measurement results appear in the photometer display 6 .

• Where applicable, use the scroll keys 7 to scroll down through the display to show other measure-
ment results.
• Click <START> 8 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

15.7 Evaluation

E*H value
Statement of the results: Δ
ΔL* value
Δa* value
Δb* value
L*a*b* values for the reference sample are shown with a lower-case “R”
(e. g. L*R)
L*a*b* values for the analysis sample are shown with a lower-case “T” (e. g. L*T)
determined for standard illuminant D65 and the 2° standard observer

15.8 Literature

1. HunterLab Application Note Vol. 8, No. 9, 06/08, www.hunterlab.com

2. ASTM E-308-06, Standard Practice for Computing the Colors of Objects by Using the CIE System

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16 HunterLab color space

16.1 Method

The HunterLab color space describes all perceivable colors and, like the CIELAB color space, is based on the
CIE’s XYZ model from 1931. This model takes the spectral characteristics of a color sample to calculate the
so-called tristimulus values X, Y, Z, which stand in direct correlation to the intensity of the stimulation of the
three types of color receptors (cone cells) of the human eye (red, green, blue). Since the XYZ model proved
to be difficult to handle in the assessment of color differences, various scientists worked on other models
that were better capable of imaging the human color perception than the XYZ model. These endeavours
resulted in the establishment of the HunterLab model in the 1960s. The HunterLab model is a cube-shaped
coordinate system with the coordinates L*, a*, and b*.
On the a* axis, green and red lie opposite each other, on the b* axis blue and yellow. Gray lies at the coor-
dinate origin (a*=0 and b*=0). The L* axis stands in the coordinate origin vertically at the a*b* level and
describes the brightness of the color, with values ranging between 0 and 100.
Negative a* values stand for green colors, positive a* values for red colors. On the b* axis, negative b* val-
ues describe blue colors, with positive b* values describing yellow colors.
The Hunter L*a*b* values are determined by measuring the transmittances in a wavelength spectrum
between 360 - 780 nm and subsequently calculating the L*a*b* values according to the methods of Hunter-
Lab Application Note Vol. 8, No. 9, 06/08 (see literature reference1).

16.2 Measuring range

Method 2583 Hunter Lab D65/2° L* 0,00 - 105,00

a* -180,0 - 180,0
b* -180,0 - 180,0
(10-mm rectangular cell)

Note
The measurement values are determined for standard illuminant D65 and the 2° standard observer.

16.3 Sample material

Clear liquid samples

Turbid liquid samples after filtration

16.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

16.5 Preparation

• Filter turbid sample solutions over a membrane filter.

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16.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2583 “Hunter Lab D65/2°”.

• Zeroing the photometer:

• Fill the sample solution into a 10-mm rectangular cell and insert the cell into the cell compartment. The
measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

6
5

The measurement results appear in the photometer display 4 .

16.7 Evaluation

Statement of the results: L

ab values
Tristimulus values X, Y, Z
determined for standard illuminant D65 and the 2° standard observer

16.8 Literature

1. HunterLab Application Note Vol. 8, No. 9, 06/08, www.hunterlab.com

2. ASTM E-308-06, Standard Practice for Computing the Colors of Objects by Using the CIE System

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17 ICUMSA Color GS1/3-7

17.1 Method

The color of sugars and its solutions is generally determined using theofficial methods of the ICUMSA (Inter-
national Commission for Uniform Methods of Sugar Analysis). The ICUMSA has described detailed proce-
dures for various types of sugar and different degrees of color.
The ICUMSA Color GS1/3-7 method is one for the determination of the color of solutions of raw sugar,
brown sugar, or sugar syrup.
In the analysis, the sample under investigation is dissolved in water and adjusted to a pH of 7.0. Subse-
quently the dry mass is measured by refractometry and the absorption of the prepared sample is deter-
mined photometrically at 420 nm, with the results being used to calculate the ICUMSA color.
The ICUMSA color is expressed in ICUMSA units at pH 7.0 (IU7.0).
The method is analogous to the ICUMSA Method Book GS1/3-7 (see literature reference1).

17.2 Measuring range

Method 2548 ICUMSA Color GS1/3-7 0 - 50 000 IU7.0

(10-mm rectangular cell)

0 - 25 000 IU7.0
(20-mm rectangular cell)

0 - 10 000 IU7.0
(50-mm rectangular cell)

17.3 Sample material

Sugar with a color index > 250 IU7.0 (raw sugar, strongly colored white plantation sugar, partly refined
brown sugar, sugar syrup)

17.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114946 - Rectangular cells 10 mm and/or
Cat. No. 114947 - Rectangular cells 20 mm and/or
Cat. No. 114944 - Rectangular cells 50 mm
ater for analysis EMSURE® or
• Cat. No. 116754 - W
distilled water
• Cat. No. 109060 - Hydrochloric acid 0,1 mol/l Titripur®
• Cat. No. 109141 - Sodium hydroxide solution 0,1 mol/l Titripur®
• Membrane filters made of cellulose nitrate, pore size 0.45 µm
• pH-Meter
• Refractometer
• Ultrasound bath
• 250-ml conical flasks
• 500-ml volumetric flasks
Standard laboratory glassware (e. g. glass beakers) and pipettes

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17.5 Preparation

• Homogenize the sample.

• Weigh the sample and water for analysis into a conical flask as per the instructions in “Table 1” and dis-
solve by swirling at room temperature.
• Adjust the pH of the sample to 7.0 ± 0.1 with hydrochloric acid 0.1 mol/l or sodium hydroxide solution
0.1 mol/l.
• Vacuum-filter the solution over a membrane filter.
• Degas the filtered solution in an ultrasound bath for 3 min.
• Use a refractometer to determine the dry mass of the solution in % RDS (refractometric dry substance)
with an accuracy of ± 0.1 g/100 g (acc. to ICUMSA Method GS4/3-13).

Table 1
Weights of sample and water and recommended optical path length of the cell for photometric measurement

Path length ICUMSA Color range Sample Water

[mm] [IU7.0] [g] [g]
< 250 50 ± 1 50 ± 1
50 250 - 500 30 ± 1 70 ± 1
500 - 2000 30 ± 1 70 ± 1
500 - 2000 30 ± 1 70 ± 1
20 2000 - 7000 10 ± 1 90 ± 1
7000 - 25 000 5.0 ± 0.1 95 ± 1
500 - 2000 30 ± 1 70 ± 1
2000 - 7000 10 ± 1 90 ± 1
10
7000 - 25 000 5.0 ± 0.1 95 ± 1
> 25 000 5.0 ± 0.1 95 ± 1

17.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2548 “ICUMSA Color GS1/3-7”.

• Entry of the RDS factor:

4
5
2

• Enter the RDS factor of the sample in %, accurate to 0.1 % 1 , confirm with “OK” 2 , and change to
the measurement procedure by clicking on “START” 3 .

• Zeroing the photometer:

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Note
Alternatively, the zero-adjustment procedure can be carried out before the RDS factor is entered. Do this by
clicking on “X” 4 to close the input mask and then click on the key “Settings” 5 . This opens a window
with various selection options.
Subsequently click on the key “ZERO ADJUSTMENT”.

• For the zero adjustment fill a 10-mm rectangular cell with water for analysis.
• After prompting, insert the filled rectangular cell into the cell compartment. The zero adjustment is
performed automatically.
• Confirm the performance of the zero-adjustment procedure by clicking on <OK> 6 .
The zero adjustment is valid for the entire measurement series.

• Fill the sample solution into a 10-mm rectangular cell and insert the cell into the cell compartment. The
measurement is performed automatically.

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A () appears behind the cue “Insert sample” 7 .

• Confirm the measurement by clicking on <OK> 8 .

The measurement result in the units IU7.0 appears in the photometer display. 9 .
• Click<START> 10 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

Note
According to ICUMSA Method GS1/3-7 the method checks automatically if the measured absorption is below
2.500 A. If the absorption is not in this range, a pop-up message appears: “Condition not met – Absorbance
> 2.5”.
In these cases, the sample preparation needs to be adjusted. Use an appropriate cell length, sample and
water weight according to recommendations of Table 1.

17.7Evaluation
Statement of the results: I CUMSA units at pH 7.0 (IU7.0)

17.8 Literature

1. ICUMSA Methods Book 2013 - ISBN 978-3-87040-555-7, Method ICUMSA Color GS1/3-7

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18 ICUMSA Color GS2/3-9

18.1 Method

The color of sugars and sugar solutions is generally determined using the official methods of the ICUMSA
(International Commission for Uniform Methods of Sugar Analysis). The ICUMSA has described detailed pro-
cedures for various types of sugar and different degrees of color.
The ICUMSA Color GS2/3-9 method is one for the determination of the color of solutions of white sugar or
highly pure sugar syrup with a color index up to 600 IU.
In the analysis, the sample under investigation is dissolved in a buffer with a pH of 7.0.
Subsequently the dry mass is measured by refractometry and the absorption of the prepared sample is
determined photometrically at 420 nm, with the results being used to calculate the ICUMSA color.
The ICUMSA color is expressed in ICUMSA units at pH 7.0 (IU7.0).
The method is analogous to the ICUMSA Method Book GS2/3-9 (see literature reference1).

18.2 Measuring range

Method 2549 ICUMSA Color GS2/3-9 0 - 600 IU7.0

(50-mm rectangular cell)

0 - 600 IU7.0
(100-mm rectangular cell,
only Prove 600)

18.3 Sample material

Sugar with a color index up to 600 IU7.0 (crystalline white sugar, icing sugar, sugar syrup)

18.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114944 - Rectangular cells 50 mm and/or
Cat. No. 174011 - Rectangular cells 100 mm (only for Prove 600)
ater for analysis EMSURE® or
• Cat. No. 116754 - W
distilled water
• Cat. No. 109060 - Hydrochloric acid 0,1 mol/l Titripur®
• Cat. No. 108379 - Triethanolamine (TEA) GR for analysis
• Membrane filters made of cellulose nitrate, pore size 0.45 µm
• pH-Meter
• Refractometer
• Ultrasound bath
• 250-ml conical flasks
• 500-ml volumetric flasks
Standard laboratory glassware (e. g. glass beakers) and pipettes

18.5 Preparing the solutions

• Preparing TEA buffer solution (pH 7.0):

Dissolve 7.46 g triethanolamine (TEA) GR in 200 ml water for analysis
Transfer to a 500 ml volumetric flask
Make up to the mark with water for analysis and mix
Transfer the entire contents of the voluimetric flask to a 1000 ml glass beaker and adjust to pH 7.0 with
hydrochloric acid 0.1 mol/l (approx. 420 ml hydrochloric acid 0.1 mol/l is required)

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18.6 Preparation

• H omogenize the sample.

• Weigh 50.0 ± 0.1 g of sample into a conical flask.
Weigh in 50.0 ± 0,1 g of TEA buffer solution (pH 7.0) and dissolve by swirling at room temperature
• Vacuum-filter the solution over a membrane filter.
• Degas the filtered solution in an ultrasound bath for 3 min.
• Use a refractometer to determine the dry mass of the solution in % RDS (refractometric dry substance)
with an accuracy of ± 0.1 g/100 g (acc. to ICUMSA Method GS4/3-13).

18.7 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2549 “ICUMSA Color GS2/3-9”.

• Entry of the RDS factor:

4
5
2

• Enter the RDS factor of the sample in %, accurate to 0.1% 1 , confirm with “OK” 2 , and change to
the measurement procedure by clicking on “START” 3 .

• Zeroing the photometer:

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• Fill the sample solution into a corresponding rectangular cell and insert the cell into the cell compart-
ment. The measurement is performed automatically.

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A () appears behind the cue “Insert sample” 7 .

• Confirm the measurement by clicking on <OK> 8 .

The measurement result in the units IU7.0 appears in the photometer display 9 .
• Click <START> 10 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

18.8 Evaluation

Statement of the results: I CUMSA units at pH 7.0 (IU7.0)

18.9 Literature

1. ICUMSA Methods Book 2013 - ISBN 978-3-87040-555-7, Method ICUMSA Color GS2/3-9

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19 ICUMSA Color GS2/3-10

19.1 Method

The color of sugars and its solutions is generally determined using the official methods of the ICUMSA
(International Commission for Uniform Methods of Sugar Analysis). The ICUMSA has described detailed pro-
cedures for various types of sugar and different degrees of color.
The ICUMSA Color GS2/3-10 method is one for the determination of the color of solutions of white sugar
with a color index up to 50 IU.
In the analysis, the sample under investigation is dissolved in water.
Subsequently the dry mass is measured by refractometry and the absorption of the prepared sample is
determined photometrically at 420 nm, with the results being used to calculate the ICUMSA color.
The method is analogous to the ICUMSA Method Book GS2/3-10 (see literature reference1).

19.2 Measuring range

Method 2550 ICUMSA Color GS2/3-10 0 - 50 IU

(50-mm rectangular cell)

0 - 50 IU
(100-mm rectangular cell,
only Prove 600)

19.3 Sample material

Sugar with a color index up to 50 IU (crystalline white sugar, icing sugar, sugar syrup)

19.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114944 - Rectangular cells 50 mm and/or
Cat. No. 174011 - Rectangular cells 100 mm (only for Prove 100)
ater for analysis EMSURE® or
• Cat. No. 116754 - W
distilled water
• Membrane filters made of cellulose nitrate, pore size 0.45 µm
• Refraktometer
• Ultrasound bath
• 250-ml conical flasks
Standard laboratory glassware (e. g. glass beakers) and pipettes

19.5 Preparation

• Homogenize the sample.

• Weigh 50.0 ± 0.1 g of sample into a conical flask.
Weigh in 50.0 ± 0,1 g of ater for analysis and dissolve by swirling at room temperature.
• Vacuum-filter the solution over a membrane filter.
• Degas the filtered solution in an ultrasound bath for 3 min.
• Use a refractometer to determine the dry mass of the solution in % RDS (refractometric dry substance)
with an accuracy of ± 0.1 g/100 g (acc. to ICUMSA Method GS4/3-13).

19.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2550 “ICUMSA Color GS2/3-10”.

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• Entry of the RDS factor:

4
5
2

• Enter the RDS factor of the sample in %, accurate to 0.1% 1 , confirm with “OK” 2 , and change to
the measurement procedure by clicking on “START” 3 .

• Zeroing the photometer:

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• Fill the sample solution into a corresponding rectangular cell and insert the cell into the cell compart-
ment. The measurement is performed automatically.

A () appears behind the cue “Insert sample” 7 .

• Confirm the measurement by clicking on <OK> 8 .

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The measurement result in the units IU appears in the photometer display 9 .

<START> 10 to start the measurement procedure for the next sample.
• Click
A renewed zero adjustment is not necessary.

19.7 Evaluation

Statement of the results: I CUMSA units (IU)

19.8 Literature

1. ICUMSA Methods Book 2013 - ISBN 978-3-87040-555-7, Method ICUMSA Color GS2/3-10

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20 ICUMSA Color GS9/1/2/3-8

20.1 Method

The color of sugars and its solutions is generally determined using the official methods of the ICUMSA
(International Commission for Uniform Methods of Sugar Analysis). The ICUMSA has described detailed pro-
cedures for various types of sugar and different degrees of color.
The ICUMSA Color GS9/1/2/3-8 method is one for the determination of the color of solutions of raw sugar,
white sugar, brown sugar, or plantation sugar.
In the analysis, the sample under investigation is dissolved in water and a buffer with a pH of 7.0.
Subsequently the absorption of the prepared sample is determined photometrically at 420 nm, with the
results being used to calculate the ICUMSA color.
The ICUMSA color is expressed in ICUMSA units at pH 7.0 (IU7.0).
The method is analogous to the ICUMSA Method Book GS9/1/2/3-8 (see literature reference1).

20.2 Measuring range

Method 2551 ICUMSA Color GS9/1/2/3-8 0 - 20 000 IU7.0

(10-mm rectangular cell)

0 - 10 000 IU7.0
(20-mm rectangular cell)

0 - 4 000 IU7.0
(50-mm rectangular cell)

20.3 Sample material

Sugar with a color index up to 16,000 IU7.0 (raw sugar, plantation white sugar, refined raw sugar)

20.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114946 - Rectangular cells 10 mm and/or
Cat. No. 114947 - Rectangular cells 20 mm and/or
Cat. No. 114944 - Rectangular cells 50 mm
ater for analysis EMSURE® or
• Cat. No. 116754 - W
distilled water
• Cat. No. 109137 - Sodium hydroxide solution 1 mol/l, Titripur®
• Cat. No. 475898 - MOPS, free Acid, ULTROL® grade
• Cat. No. SLHA025NB - M
embrane filters Millex-HA, 0,45 µm pore size, 25 mm diameter,
Luer-Lock fitting inlet
• Cat. No. SLAA025NB - M
embrane filters prefilter unit, 0,80 µm pore size, 25 mm diameter,
Luer-Lock fitting inlet
• Cat. No. XX1102012 Plastic syringes 10 - 50 ml with Luer-Lock fitting outlet
• pH-Meter
• Refractometer
• Ultrasound bath
• 100-ml and 1000-ml volumetric flasks
• 250-ml conical flasks
Standard laboratory glassware (e. g. glass beakers) and pipettes

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20.5 Preparing the solutions

• Preparing MOPS buffer solution:

Dissolve 41.8 g MOPS in 800 ml water for analysis
Adjust to pH 7.00 ± 0.01 with sodium hydroxide solution 1 mol/l
Transfer to a 1000-ml volumetric flask
Make up to the mark with water for analysis and mix

• Preparing the reference solution for the zero-adjustment procedure:

Transfer 10.0 ml MOPS buffer solution to a 100 ml volumetric flask
Make up to the mark with water for analysis and mix

20.6 Preparation

• Homogenize the sample.

• Weigh the sample and water for analysis into a conical flask as per the instructions in “Table 1” and note
the sample weight.
• Add 60 - 70 ml of water and and dissolve by swirling at room temperature.
• Transfer to a 100-ml volumetric flask.
• Add 10.0 ml of MOPS buffer solution and make up the contents of the volumetric flask to the mark with
water for analysis.
• Fill a plastic syringe with 10 - 20 ml of the pretreated sample solution, attach a membrane filter to the
syringe, and subsequently connect the Millex-HA syringe filter unit to the prefilter.
• Filter the sample solution into a clean and dry glass beaker.
• Degas the filtered solution in an ultrasound bath for 3 min.

Table 1
Weights of sample and water and recommended optical path length of the cell for photometric measurement

Path length ICUMSA Color range Sample

[mm] [IU7.0] [g]
0 - 1000 20.0 ± 0.1
50 200 - 2000 10.0 ± 0.1
400 - 4000 5.0 ± 0.1
250 - 2500 20.0 ± 0.1
20 500 - 5000 10.0 ± 0.1
1000 - 10 000 5.0 ± 0.1
500 - 5000 20.0 ± 0.1
10 1000 - 10 000 10.0 ± 0.1
2000 - 20 000 5.0 ± 0.1

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20.7 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2551 “ICUMSA Color GS9/1/2/3-8”.

• Entering the sample weight:

4
5
2

• Enter the weight of the sample in grams (g), accurate to 0.1 grams (g) 1 , confirm with “OK” 2 , and
change to the measurement procedure by clicking on “START” 3 .

• Zeroing the photometer:

• The photometer must be zeroed prior to each new measurement series.
• It is advisable to zero the photometer using the same cell as the one used for the measurement samp-
le or else a cell with identical optical characteristics and an identical absorption (matched pair).
The zero-adjustment procedure for the measurement series is automatically prompted by the instru-
ment after the entry of the weight of the first analytical sample.

Note
Alternatively, the zero-adjustment procedure can be carried out before the sample weight is entered. Do
this by clicking on “X” 4 to close the input mask and then click on the key “Settings” 5 . This opens a win-
dow with various selection options.
Subsequently click on the key “ZERO ADJUSTMENT”.

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• Fill the sample solution into a corresponding rectangular cell and insert the cell into the cell compart-
ment. The measurement is performed automatically.

A () appears behind the cue “Insert sample” 7 .

• Confirm the measurement by clicking on <OK> 8 .

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Note
According to ICUMSA Method GS9/1/2/3-8 the method checks automatically if the measured absorption is
below 2.500 A. If the absorption is not in this range, a pop-up message appears: “Condition not met –
Absorbance > 2.5”.
In these cases, the sample preparation needs to be adjusted. Use an appropriate cell length, sample and
water weight according to recommendations of Table 1.

20.8 Evaluation

Statement of the results: I CUMSA units at pH 7.0 (IU7.0)

20.9 Literature

1. ICUMSA Methods Book 2013 - ISBN 978-3-87040-555-7, Method ICUMSA Color GS9/1/2/3-8

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21 Iodine color number

21.1 Method

The iodine color number is used to assess the color of clear liquids that possess color characteristics similar
to those of the iodine color scale.
The method was originally based on the visual comparison of the color of the sample solution with defined
iodine-color standards. Meanwhile, however, in practice the photometric principle has become the predomi-
nantly used method for determining the iodine color, since it enables a substantially higher degree of preci-
sion to be achieved.
The measurement is made in the wavelength spectrum in which the yellow to yellow-brown standard solu-
tions of the iodine color scale absorb light. The measurement results are referenced to the respective mea-
surement conditions and are traceable to a defined color standard prepared from iodine and potassium
iodide in aqueous solution.
The “iodine color number” method can be performed in the Prove spectrophotometers at the wavelengths
445 nm and 340 nm. At the 445 nm wavelength, the typical range of applications of the method – the
assessment of the color of clear liquids, e. g. solvents, resins, oils, and fatty acids – shows a good compara-
bility of the photometric method with the visual method. At 340 nm – a wavelength that is still just within
the so-called VIS spectrum, but is no longer visible to the human eye – the iodine color number standards
show their highest absorption in the VIS spectrum. This wavelength can be practically used for the assess-
ment of weakly colored samples. Alternatively, weakly colored samples can also be analyzed using the Pt/Co
color index (see “Color Hazen”, section 10).
The methods are analogous to DIN 6162 (see literature reference1), DGF C-IV 4 a (see literature refer-
ence2), DGF F-I 2 (see literature reference3), and DGF H-II 3 (see literature reference4).

21.2 Measuring range

Method 21 Iodine color number 445 0.2 - 10.0

(50-mm rectangular cell)

0.5 - 25.0
(20-mm rectangular cell)

1.0 - 50.0
(10-mm rectangular cell)

Method 33 Iodine color number 340 0.010 - 0.600

(50-mm rectangular cell)

0.03- 1.50
(50-mm rectangular cell)

0.05 - 3.00
(50-mm rectangular cell)

21.3 Sample material

Clear liquid samples with color characteristics similar to those on the iodine color number scale (= yellow to
yellow-brown)
Turbid liquid samples with color characteristics after filtration similar to those on the iodine color number
scale (= yellow to yellow-brown)
e. g. solvents, plasticizers, resins, resin solutions, oils, fatty acids, lecithins, surfactants

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21.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

21.5 Preparation

• Filter turbid sample solutions over a membrane filter.

To do this, prerinse the membrane filter with approx. 50 ml of water for analysis, then rerinse with
approx. 50 ml of the sample solution and discard the filtrate.
Filter a further 50 ml of the sample solution over the prerinsed membrane filter and use the filtrate for
the analysis.
• Prior to the determination, heat solid samples to a few degrees Kelvin above melting point, homogenize
by stirring gently, taking care to avoid air bubbles. Analyze the sample immediately.
• Where necessary, homogenize lecithins by gentle melting.
Dissolve 10 g of the sample in 70 ml of toluene, transfer to a 100 ml volumetric flask, amd make up to
the mark with toluene. Use this solution for the analysis, expressing the result for a 10% solution.

21.6 Procedure and measurement

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 21 “Iodine color number 445” resp. No. 33 “Iodine color number 340”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

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If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• Open the methods list (<Methods>) 4 and select Method No. 21 “Iodine color number 445”
resp. No. 33 “Iodine color number 340”.

21.7 Evaluation

Statement of the results: I FZ (= Iodfarbzahl (Iodine color number))

21.8 Method control

The method can be checked using iodine color number standard according to the DIN 6162 standard.

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21.9 Recalibration

The methods can be recalibrated using iodine color number standards according to the DIN standards.
Recalibration can be done by measuring value pairs, by entering value pairs, or by entering coefficients of a
calibration function. Please refer to the Operating Manual of your spectrophotometer for details on the recal-
ibration procedure.

21.10 Literature

1. DIN 6162:2014-09 - Determination of iodine colour number

2. DGF Einheitsmethoden (2. Auflage einschließlich 14. Akt.-Lfg von 2009) - Abteilung Fette - C-IV 4 -
Farbmessung
3. DGF Einheitsmethoden (2. Auflage einschließlich 14. Akt.-Lfg von 2009) - Abteilung Fettbegleitstoffe -
F-I 2 - Farbe von Lecithinen
4. DGF Einheitsmethoden (2. Auflage einschließlich 14. Akt.-Lfg von 2009) - Abteilung Tenside - H-II 3 - Farbe

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22 Klett color index

22.1 Method

The Klett color index is one of the oldest known color indices. It has been measured using special filter pho-
tometers, the Klett Summerson colorimeters, since about 1930 right up to the present day. These photome-
ters use exchangeable filters, each with characteristic transmittances in various wavelength spectrums. The
most frequently used filter is the KS-42 blue filter, for the measurement of yellow colors. The filter allows
light to pass through in the 400 - 450 nm wavelength spectrum, showing its greatest light transmission at
about 420 nm. The Klett color scale itself is based on the measured absorption in the wavelength spectrum
of the filter being used. The Klett color number is calculated by multiplying the measured absorption by a
factor of 500. The optical path length of the cell used is not used in the calculation. The Klett color scale is
defined for the range 0 - 1000.
In modern-day spectrophotometers, the Klett color number is frequently measured at the wavelength of the
highest light transmission of the corresponding filter. For the KS-42 filter, this is at 417 nm. In this case, the
Klett color number is determined using an empirically defined factor. In practice, this factor is determined by
comparatively measuring the absorption of a platinum/cobalt color-reference solution at the selected wave-
length of the spectrophotometer (417 nm) and also determining the Klett color index yielded by a
Klett-Summerson colorimeter using a KS-42 filter. Care must be taken to ensure that the results of both
methods for colors of the Pt/Co scale are absolutely comparable. If samples with deviating colors for the
platinum/cobalt solution are analyzed, there may be slight differences in the results yielded by measure-
ment at 417 nm and by the use of the KS-42 filter. For example: the Klett color index of a yellow chromate
solution measured at 417 nm is roughly 5% below the result measured with the KS-42 filter.
The Spectroquant® Prove spectrophotometers determine the Klett color index by measuring the absorption
at the 417 nm wavelength and subsequent calculation of the Klett color index. The result is expressed in the
unit Klett417. The optical path length of the cell used is also given as a citation form.

22.2 Measuring range

Method 311 Klett Color 0 - 1000 Klett417

(50-mm rectangular cell)

22.3 Sample material

Clear yellow to yellow-brown liquids

22.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114944 - Rectangular cells 50 mm
ater for analysis EMSURE® or
• Cat. No. 116754 - W
distilled water
• Ultrasound bath (optional)
• Heating bath (optional)
• Filter paper or centrifuge (optional)

22.5 Preparation

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22.6 Procedure and measurement

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 311 “Klett Color”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• For the zero adjustment fill a 50-mm rectangular cell with water for analysis.
• After prompting, insert the filled rectangular cell into the cell compartment. The zero adjustment is
performed automatically.
• Confirm the performance of the zero-adjustment procedure by clicking on <OK> 3 .

• Open the methods list (<Methods>) 4 and select Method No. 311 “Klett Color”.

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• Fill the sample solution into a 50-mm rectangular cell and insert the cell into the cell compartment..
The measurement is performed automatically and the result appears in the display 5 .

22.7 Evaluation

Statement of the results: Klett417, 50 mm

22.8 Literature

none

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23 Saybolt color measurement

23.1 Method

The Saybolt color scale serves the assessment of the color of clear, liquid petroleum products. The scale
ranges from -16 (darkest color) to +30 (lightest color).
The value is determined by measuring the transmittances in the wavelength spectrum between 380 - 780
nm and subsequent calculation of the tristimulus values X, Y, Z. In further calculation steps according to the
ASTM D6045 standard (see literature reference1) the tristimulus values are taken as a basis for the calcula-
tion of the Saybolt color number.
The method is analogous to ASTM D6045 (see literature reference1).

23.2 Measuring range

Method 2563 Saybolt Color 50 mm -16,0 - 31,0 Saybolt

(50-mm rectangular cell)

Method 2564 Saybolt Color 100 mm -16,0 - 31,0 Saybolt

(100-mm rectangular cell,
only Prove 600)

23.3 Sample material

Clear yellow to yellow-brown liquids

Products made of natural resins (e.g. tall oils, tall-oil fatty acids, paints, oils)
Surfactants

23.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

23.5 Preparation

23.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2563 resp. 2564 “Saybolt Color”.

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Zeroing the photometer:

• Fill the sample solution into a corresponding rectangular cell and insert the cell into the cell compart-
ment. The measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

The measurement result appears in the photometer display 4 .

• Click <START> 5 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

23.7 Evaluation

Statement of the results: Saybolt Units

23.8 Literature

1. ASTM D6045-12, Standard Test Method for Color of Petroleum Products by the Automatic Tristimulus Method

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24 Spectral absorption coefficient, SAC α(254)

24.1 Method

The spectral absorption coefficient at 254 nm is a parameter for the degree of water pollution caused by dis-
solved organic substances. These substances include inter alia humic matter from the soil, aromatic com-
pounds, and metabolic products of microorganisms.
The spectral absorption coefficient is measured by transmitting light with a wavelength of 254 nm through a
filtered sample.
The spectral absorption coefficient at 254 nm is expressed in the unit m-1 and in accordance with the defini-
tion of the DIN 38404-3 standard is based on the absorption of UV light caused by organic matter dissolved
in the sample.
The method is analogous to DIN 38404-3 (see literature reference1).

24.2 Measuring range

Method 300 SAC α(254) 0.1 - 50.0 m-1

(50-mm rectangular cell quartz)

0.3 - 125.0 m-1

(20-mm rectangular cell quartz)

1 - 250 m-1
(10-mm rectangular cell quartz)

24.3 Sample material

Water from water-treatment plants (crude water, drinking water, weakly colored industrial wastewater)
Surface water

24.4 Reagents and auxiliaries

• Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or

24.5 Preparation

• Filter turbid sample solutions over a membrane filter.

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24.6 Procedure and measurement

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 300 “SAC α(254)”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• For the zero adjustment fill a corresponding rectangular quartz cell with water for analysis.
• After prompting, insert the filled rectangular cell into the cell compartment. The zero adjustment is
performed automatically.
• Confirm the performance of the zero-adjustment procedure by clicking on <OK> 3 .

• Open the methods list (<Methods>) 4 and select Method No. 300 “SAC α(254)”.

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• Fill the sample solution into a corresponding rectangular quartz cell and insert the cell into the cell
compartment. The measurement is performed automatically and the result appears in the display 5 .

24.7 Evaluation

Statement of the results: m

-1

24.8 Literature

1. DIN 38404-3:2005-07 - German standard methods for the examination of water, waste water and sludge
- Physical and physicalchemical parameters (group C) - Part 3: Determination of absorption in the range
of the ultraviolet radiation, Spectral absorptions coefficient (C 3)

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25 Spectral absorption coefficient, SAC α(436)

25.1 Method

The spectral absorption coefficient at 436 nm is a parameter of the German Drinking Water Ordinance and
the EU Drinking Water Directive. This parameter is used to assess yellow to yellow-brown colors (true color)
of a filtered water sample. These discolorations of a water sample are in most cases due to dissolved humic
matter and also to iron and manganese compounds.
The method is analogous to German Drinking Water Ordinance, Annex 3, No. 7 (see literature reference1)
and the EU Drinking Water Directive 98/83/EU (see literature reference2). The method is analogous to a part
of DIN EN ISO 7887 - Method B (see literature reference3).

25.2 Measuring range

Method 302 SAC α(436) 0.1 - 50.0 m-1

(50-mm rectangular cell)

0.3 - 125.0 m-1

(20-mm rectangular cell)

1 - 250 m-1
(10-mm rectangular cell)

25.3 Sample material

Water from water-treatment plants (crude water, drinking water, weakly colored industrial wastewater)

25.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

25.5 Preparation

• Filter turbid sample solutions over a membrane filter.

25.6 Procedure and measurement

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 302 “SAC α(436)”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

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If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• Open the methods list (<Methods>) 4 and select Method No. 302 “SAC α(436)”.

25.7 Evaluation

Statement of the results: m

-1

25.8 Literature

1. Deutsche Trinkwasserverordnung - TrinkwV Anlage 3, No. 7

2. EU-Trinkwasserrichtlinie 98/83/EG
3. DIN EN ISO 7887:2011 - Water quality - Examination and determination of colour (ISO 7887:2011); Ger-
man version EN ISO 7887:2011

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26 Spectral attenuation coefficient, SAC µ(254)

26.1 Method

The spectral attenuation coefficient at 254 nm for the degree of water pollution caused by particulate matter
and dissolved organic substances. These substances include inter alia suspended matter such as solid min-
eral or organic substances, sediment matter, humic matter from the soil, aromatic compounds, and meta-
bolic products of microorganisms.
The spectral attenuation coefficient is measured by transmitting light with a wavelength of 254 nm through
a filtered sample.
The spectral attenuation coefficient at 254 nm is expressed in the unit m-1 and in accordance with the defi-
nition of the DIN 38404-3 standard is calculated from the sum of the absorption and the diffusion of the UV
light caused by particulate matter and organic matter dissolved in the sample.
The method is analogous to the DIN 38404-3 standard (see literature reference1).

26.2 Measuring range

Method 301 SAC µ(254) 0.1 - 50.0 m-1

(50-mm rectangular cell quartz)

0.3 - 125.0 m-1

(20-mm rectangular cell quartz)

1 - 250 m-1
(10-mm rectangular cell quartz)

26.3 Sample material

Water from water-treatment plants (crude water, drinking water, weakly colored industrial wastewater)
Surface water

26.4 Reagents and auxiliaries

• Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or

26.5 Preparation

•
Shake the unfiltered sample to resuspend any suspended or turbidity-causing matter evenly in the sample.
• Measure the sample immediately after resuspension.

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26.6 Procedure and measurement

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 301 “SAC µ(254)”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• Open the methods list (<Methods>) 4 and select Method No. 301 “SAC µ(254)”.

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26.7 Evaluation

Statement of the results: m

-1

26.8 Literature

1. DIN 38404:2005-07 - German standard methods for the examination of water, waste water and sludge -
Physical and physicalchemical parameters (group C) - Part 3: Determination of absorption in the range of
the ultraviolet radiation, Spectral absorptions coefficient (C 3)

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27 Spectral attenuation coefficient, SAC µ(254) - corrected

27.1 Method

The corrected spectral attenuation coefficient at 254 nm is a parameter for the degree of water pollution
caused by dissolved organic substances. These substances include inter alia humic matter from the soil,
aromatic compounds, and metabolic products of microorganisms.
The spectral absorption coefficient is measured by transmitting light with a wavelength of 254 nm through
an unfiltered and uncolored sample.
Any particulate matter present in the sample, e. g. solid mineral or organic substances or sediment matter,
result in an additional attenuation of the UV light. To compensate the contribution of such matter to the
spectral attenuation coefficient, in a further measurement light with a wavelength of 550 nm is transmitted
through the sample. Light with this wavelength is attenuated by particulate matter present in the sample in
a similar way as light with a wavelength of 254 nm, but is not absorbed by dissolved organic matter.
The corrected spectral attentuation coefficient at 254 nm is expressed in the unit m-1 and in accordance with
the definition of the DIN 38404-3 standard is based on the attenuation of UV light with a wavelength of 254
nm, corrected by the contribution of light with a wavelength of 550 nm.
The method is analogous to the DIN 38404-3 standard (see literature reference1).

27.2 Measuring range

Method 2571 SAC µ(254), korr 0.1 - 50.0 m-1

(50-mm rectangular cell quartz)

0.3 - 125.0 m-1

(20-mm rectangular cell quartz)

1 - 250 m-1
(10-mm rectangular cell quartz)

27.3 Sample material

Uncolored water from water-treatment plants (crude water, drinking water, weakly colored industrial waste-
water)
Uncolored surface water

27.4 Reagents and auxiliaries

• Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or

27.5 Preparation

•
Shake the unfiltered sample to resuspend any suspended or turbidity-causing matter evenly in the sample.
• Measure the sample immediately after resuspension.

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27.6 Procedure and measurement

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 2571 “SAC µ(254), korr”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• Open the methods list (<Methods>) and select Method No. 2571 “SAC µ(254), korr”.

• Fill the sample solution into a corresponding rectangular quartz cell and insert the cell into the cell
compartment. The measurement is performed automatically.

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A () appears behind the cue “Insert cell” 4 .

• Confirm the measurement by clicking on <OK> 5 .

The measurement result appears in the photometer display 6 .

27.7 Evaluation

Statement of the results: m

-1

27.8 Literature

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28 Tint index
28.1 Method

The tint index is frequently determined together with the whiteness of a material (see also section 32,
“Whiteness”). In materials that are not colored, the color index lies at “0”. Greenish colors are presented as
values in the positive range, reddish colors as ones in the negative range. The higher the value, the stron-
ger the color (color cast).
The tint index is determined by measuring the transmittances of the sample in a wavelength spectrum
between 380 and 780 nm and subsequent calculation of the tristimulus values X, Y, Z. The tristimulus val-
ues are then used to calculate the color index according to the methods of the ASTM E313-15 standard.
The method is analogous to the ASTM E313-15 method (see literature reference1).

28.2 Measuring range

Method 2577 Tint Index -6.00 - 3.00 TI10mm

(10-mm rectangular cell)

Method 2578 Tint Index -6.00 - 3.00 TI50mm

(50-mm rectangular cell)

Note
The tint index is determined for standard illuminant C and the 2° standard observer.

28.3 Sample material

Clear liquid samples

Turbid liquid samples after filtration

28.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

28.5 Preparation

• Filter turbid sample solutions over a membrane filter.

28.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2577 resp. 2578 “Tint Index”.

• Zeroing the photometer:

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• Fill the sample solution into a corresponding rectangular cell and insert the cell into the cell compart-
ment. The measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

6
5

The measurement results appear in the photometer display 4 .

28.7 Evaluation

Statement of the results: T

int Index (TI)
Tristimulus values X, Y, Z
Chromaticity coordinates x, y
determined for standard illuminant C and the 2° standard observer

28.8 Literature

1. ASTM E313-15, Standard Practice for Calculating Yellowness and Whiteness Indices from Instrumentally
Measured Color Coordinates

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29 Transmittances TX, TY, TZ

29.1 Method

The transmittances TX, TY, TZ serve the colorimetric characterization of clear liquids. The transmittances TX,
TY, TZ were originally determined with broadband X, Y, Z standard color filters and simple filter photometers.
Today they are generally determined by spectrophotometry.
The values are determined by measuring the transmittances in the wavelength spectrum between
380 - 780 nm and subsequent calculation of the transmittances TX, TY, TZ according to the methods of
the DIN EN 1557 standard.
The method is analogous to the DIN EN 1557 standard (see literature reference1).

29.2 Measuring range

Method 2579 Transmittances TX, TY, TZ 0,0 - 150,0

(10-mm rectangular cell)

Note
The Transmittances TX, TY, TZ is determined for standard illuminant C and the 2° standard observer.

29.3 Sample material

Clear liquid samples

Turbid liquid samples after filtration

29.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

29.5 Preparation

• Filter turbid sample solutions over a membrane filter.

29.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2579 “Transmittances TX, TY, TZ”.

• Zeroing the photometer:

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• Fill the sample solution into a 10-mm rectangular cell and insert the cell into the cell compartment. The
measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

6
5

The measurement results appear in the photometer display 4 .

29.7 Evaluation

ransmittances TX, TY, TZ

Statement of the results: T
Tristimulus values X, Y, Z
Chromaticity coordinates x, y
determined for standard illuminant C and the 2° standard observer

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29.8 Literature

1. DIN EN 1557:1997-03, Surface active agents - Colorimetric characterization of optically clear coloured
liquids (products) as X, Y, Z tristimulus values in transmission
2. DIN 5033-2:1992-05, Colorimetry; Standard colorimetric systems
3. DIN 5033-3:1992-07, Colorimetry; Colorimetric measures
4. ISO 7724-1:1984-10, Paints and varnishes - Colorimetry - Part 1: Principles
5. ISO CIE 10527:1991-12, CIE standard colorimetric observers
6. DIN EN ISO 11664-1:2011-07, Colorimetry - Part 1: CIE standard colorimetric observers
(ISO 11664-1:2007); German version EN ISO 11664-1:2011

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30 UV-absorbing organic matters (UV absorption 254)

30.1 Method

Certain types of organic matter dissolved in water, for example humic matter from the soil, aromatic com-
pounds such as lignin or tannin from the timber-processing industry, metabolic products of microorganisms,
or other aromatic compounds are capable of absorbing UV light. The measurement of the absorption in the
UV spectrum can accordingly be an indicator of the possible presence of such substances in a water sample.
Besides the measurement of the spectral absorption coefficient 254 nm according to DIN 38404-3 (see sec-
tion 24, “Spectral absorption coefficient, SAC α(254)”), another method frequently used to determine
UV-absorbing organic matter is the APHA 5910 method. In technical circles, this method is frequently
referred to as the UV absorption method.
In this method, UV light with a wavelength of 254 nm is transmitted through a filtered sample. The result of
the UV absorption can be expressed in a variety of units, such as A/cm or cm-1 or m-1.
The methods are analogous to APHA 5910 (see literature reference1). The description of the APHA 5910
method mentions that the pH of the sample may have an impact on the UV absorption. The risk of potential
interferences by other matter contained in the sample is also mentioned.

30.2 Measuring range

Method 309 UV Absorbing Organic 0.0000 - 0.1000 A/cm

Constituents 0.0000 - 0.1000 cm-1
0.0 - 10.0 m-1
(100-mm rectangular cell quartz,
only Prove 600)

0.000 - 0.200 A/cm

0.000 - 0.200 cm-1
0 - 20 m-1
(50-mm rectangular cell quartz)

0.000 - 0.500 A/cm

0.000 - 0.500 cm-1
0 - 50 m-1
(20-mm rectangular cell quartz)

0.000 - 1.000 A/cm

0.000 - 1.000 cm-1
0 - 100 m-1
(10-mm rectangular cell quartz)

Method 310 UV Absorption 254 0.0000 - 0.3000 A/cm

0.0000 - 0.3000 cm-1
0.00 - 30.00 m-1
(100-mm rectangular cell quartz,
only Prove 600)

0.000 - 0.600 A/cm

0.000 - 0.600 cm-1
0.0 - 60.0 m-1
(50-mm rectangular cell quartz)

0.000 - 1.500 A/cm

0.000 - 1.500 cm-1
0.0 - 150.0 m-1
(20-mm rectangular cell quartz)

0.000 - 3.000 A/cm

0.000 - 3.000 cm-1
0.0 - 300.0 m-1
(10-mm rectangular cell quartz)

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Note
The definition of the upper measuring-range limit of Method 309 is based on the recommendation given in
APHA 5910 that samples with an absorption of > 1.0 Abs should be diluted for analysis. If this absorption
limit is exceeded in the analysis procedure, when Method 309 is used the result “HI” appears in the display
(= result lies above the upper measuring-range limit). In such a case the sample must be diluted with dis-
tilled water/water for analysis and the analysis procedure repeated.
In Method 310, the upper measurement-range limit is based on a maximum value of 3.0 Abs and is corre-
spondingly higher. When Method 310 is used for determining UV-absorbing organic matter, it is advisable to
also display the absorption and, in the case of absorption values > 1.0 Abs, to follow the recommendations
of APHA 5910 regarding dilution.
Instructions on how to display the absorption during the measurement procedure and how to include the
dilution factor in the calculation of the final result can be found in the Operating Manual for the spectropho-
tometer.

30.3 Sample material

Water from water-treatment plants (crude water, drinking water, weakly colored industrial wastewater)
Surface water
Seawater

30.4 Reagents and auxiliaries

• Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or

Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 100784 - Rectangular cells 10 mm quartz and/or
Rectangular cells 20 mm quartz and/or
Rectangular cells 50 mm quartz
Rechteckküvetten 100 mm quartz
ater for analysis EMSURE® or
• Cat. No. 116754 - W
distilled water
• Cat. No. 109535 - MQuant® pH-indicator strips 0 - 14 universal indicator
• Cat. No. 109141 - Sodium hydroxide solution 0.1 mol/l Titripur® (optional)
• Cat. No. 109060 - Hydrochloric acid 0.1 mol/l Titripur® (optional)
Standard laboratory glassware (e. g. glass beakers) and pipettes

30.5 Preparation

• Filter turbid sample solutions over a membrane filter.

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30.6 Influence of foreign agents / interferences

Particulate matter and UV-absorbing inorganic matter may have an effect on the measurement of the UV
absorption. Any particulate matter present in the sample must generally be removed by filtration.
UV-absorbing inorganic components may take the form of naturally occurring substances or other substanc-
es occurring in wastewater, e. g. iron-containing compounds, nitrates, nitrites, bromides, as well as com-
pounds used or occurring in wastewater-treatment processes such as ozone, chlorates, chlorites, chlora-
mines, and thiosulfates.
The presence of UV-absorbing inorganic components can be tested by recording an absorption spectrum of
the sample in the 200 - 400 nm wavelength spectrum. Such compounds generally appear in the spectrum
as relatively sharp peaks.
UV-absorbing inorganic components, by contrast, usually produce an unformed absorption curve, with the
absorption increasing as the wavelength diminishes.

30.7 Procedure and measurement

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 309 “UV Absorbing Organic Constituents” resp. No. 310 “UV Absorption 254”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• Open the methods list (<Methods>) 4 and select Method No. 309 “UV Absorbing Organic
Constituents” resp. No. 310 “UV Absorption 254”.

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30.8 Evaluation

Statement of the results: A

/cm or
cm-1 or
m-1

30.9 Literature

1. Standard Methods for the Examination of Water and Wastewater (21th edition), APHA 5910 UV-Absorbing
Organic Constituents - B. Ultraviolet Absorption Method

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31 UV irradiation (UV absorption 254 - UV transmission 254)

31.1 Method

UV irradiation is a method used in the water-treatment sector for disinfection purposes. The method is
based on the DNA- and RNA-changing and thus reproduction-inhibiting effect of UV light of varying wave-
lengths on microorganisms.
UV irradiation is usually carried out continuously in flow-metering systems.
The success of the disinfectant effect depends on the UV irradiance and the duration of the irradiation pro-
cess and also on the quality of the water itself.
Various components present in the water, for example hardness-forming substances, iron-containing com-
pounds, halogenides, organic matter, or microorganisms, can lead to the formation of deposits on the UV
irradiation lamp. These deposits have a negative impact on the irradiance and thus on the effectivenesss of
the procedure.
Other sources of influence are turbidities present in the water, which may lead to a diffusion of the UV light,
and dissolved organic matter capable of absorbing UV light and thus of impairing the effectiveness of the
procedure.
The presence of turbidities or dissolved organic compounds can be determined by spectrophotometric mea-
surement at 254 nm. This method is widely used in the practical area to monitor any impact on the effec-
tiveness of UV irradiation measures.
In this method, UV light with a wavelength of 254 nm is transmitted through an unfiltered sample.
The measurement results are usually expressed as the transmission or absorption relative to an optical path
length of 1 cm.

31.2 Measuring range

Method 310 UV Absorption 254 0.0000 - 0.3000 A/cm

0.000 - 0.300 cm-1
0.00 - 30.00 m-1
(100-mm rectangular cell quartz,
only Prove 600)

0.000 - 0.600 A/cm

0.000 - 0.600 cm-1
0.0 - 60.0 m-1
(50-mm rectangular cell quartz)

0.000 - 1.500 A/cm

0.000 - 1.500 cm-1
0.0 - 150.0 m-1
(20-mm rectangular cell quartz)

0.000 - 3.000 A/cm

0.000 - 3.000 cm-1
0.0 - 300.0 m-1
(10-mm rectangular cell quartz)

Method 2572 UV Transmission 254 0.00 - 105.00 %T/cm

(100-mm rectangular cell quartz,
only Prove 600)

0.00 - 105.00 %T/cm

(50-mm rectangular cell quartz)

0.00 - 105.00 %T/cm

(20-mm rectangular cell quartz)

0.00 - 105.00 %T/cm

(10-mm rectangular cell quartz)

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31.3 Sample material

Water from water-treatment plants (crude water, drinking water, weakly colored industrial wastewater)
Surface water
Seawater

31.4 Reagents and auxiliaries

• Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or

31.5 Preparation

•
Shake the unfiltered sample to resuspend any suspended or turbidity-causing matter evenly in the sam-
ple.
• Measure the sample immediately after resuspension.

31.6 Procedure and measurement - Method 310 “UV Absorption 254”

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method
No. 310 “UV Absorption 254”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

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If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• Open the methods list (<Methods>) 4 and select Method No. 310 “UV Absorption 254”.

31.7 Procedure and measurement - Method 2572 “UV Transmission 254”

Zeroing the photometer:

•
• It is advisable to zero the photometer at the start of each working day, using the same cell as the one
used for the test solutions or else a cell with identical optical characteristics and an identical absorption
(matched pair).
• To perform the zero-adjustment procedure open the methods list (<Methods>) and select Method No.
2572 “UV Transmission 254”.
If there is no valid zero adjustment available for the method, the window for the zero-adjustment pro-
cedure opens automatically.

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If a valid zero adjustment is already available, the Methods window opens.

The zero-adjustment procedure must then be selected by opening <Settings> 1 and clicking on the
selection button “ZERO ADJUSTMENT” 2 .

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• Open the methods list (<Methods>) 4 and select Method No. 572 “UV Transmission 254”.
• Fill the sample solution into a corresponding rectangular quartz cell and insert the cell into the cell
compartment. The measurement is performed automatically.

A () appears behind the cue “Insert sample” 5 .

• Confirm the measurement by clicking on <OK> 6 .

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The measurement result appears in the photometer display 7 .

• Click <START> 8 to start the measurement procedure for the next sample.

31.8 Evaluation

Statement of the results: Method 310 “UV Absorption 254”: A/cm or

cm-1 or
m-1

Method 2572 “UV Transmission 254”: %T/cm

31.9 Literature

1. Ultraviolet light disinfection in the use of individual water purification devices; U.S. Army Public Health
Command; Retrieved 2014-01-08
2. The Basic Principles of UV-Disinfection of Water; Meulemans, C. C. E.; Ozone: Science & Engineering.
Vol.9, 1987, issue 4: 299-313

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32 Whiteness
32.1 Method

The whiteness is a numerical parameter for the degree of whiteness of a surface or the transmission capaci-
ty of a clear liquid. The whiteness is primarily used in connection with the testing of solid materials, e.g.
paper, to determine any changes in color. The method can also be used to record color changes in clear liq-
uids. The greater the intensity of the color of a material (color cast), the lower the whiteness value.
If any discolorations are detected, in many cases the tint index of the material is also determined (see sec-
tion 28, “Tint index”).
The whiteness value is determined by measuring the transmittances in the wavelength spectrum between
380 - 780 nm and subsequent calculation of the tristimulus values X, Y, Z. The tristimulus values are then
taken as a basis for the calculation of the whiteness value according to the methods of ASTM E313-15.
The method is analogous to ASTM E313-15 (see literature reference1).

32.2 Measuring range

Method 2575 Whiteness 40.0 - 220.0 WI10mm

(10-mm rectangular cell)

Method 2576 Whiteness 40.0 - 220.0 WI50mm

(50-mm rectangular cell)

Note
The whiteness value is determined for standard illuminant C and the 2° standard observer.

32.3 Sample material

Clear liquid samples

Turbid liquid samples after filtration

32.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

32.5 Preparation

• Filter turbid sample solutions over a membrane filter.

32.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2575 resp. 2576 “Whiteness”.

• Zeroing the photometer:

• Fill the sample solution into a corresponding rectangular cell and insert the cell into the cell compart-
ment. The measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

6
5

The measurement results appear in the photometer display 4 .

• Where applicable, use the scroll keys 5 to scroll down through the display to show other measure-
ment results.
• Click <START> 6 to start the measurement procedure for the next sample.

32.7 Evaluation

Statement of the results: W

hiteness (TI)
Tristimulus values X, Y, Z
Chromaticity coordinates x, y
determined for standard illuminant C and the 2° standard observer

32.8 Literature

1. ASTM E313-15, Standard Practice for Calculating Yellowness and Whiteness Indices from
Instrumentally Measured Color Coordinates

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33 Yellowness
33.1 Method

White and colorless materials can take on a yellow tint due to a number of external influences, e. g. light,
sunlight, temperature, moisture, or chemical reactions.
These changes in color are usually determined by measuring the so-called yellowness index of the material
in question at regular intervals. The greater the yellowing effect, the higher the yellowness index of the
material.
The yellowness index is determined by measuring the transmittances of the sample in a wavelength spectrum
between 380 and 780 nm and subsequently calculating the tristimulus values X, Y, Z. The tristimulus values
are then used to calculate the yellowness index according to the methods of the ASTM E313-15 standard.
The method is analogous to the ASTM E313-15 standard (see literature reference1).

33.2 Measuring range

Method 2573 Yellowness 0.0 - 30.0 YI10mm

(10-mm rectangular cell)

Method 2574 Yellowness 0.0 - 90.0 YI50mm

(50-mm rectangular cell)

Note
The yellowness value is determined for standard illuminant C and the 2° standard observer and is specified
only for yellow colors.

33.3 Sample material

Colorless to yellowish clear liquid samples

Colorless to yellowish turbid liquid samples after filtration

33.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Cat. No. 173017 - Spectroquant® UV/VIS Spectrophotometer Prove 300 or
Cat. No. 173018 - Spectroquant® UV/VIS Spectrophotometer Prove 600
• Cat. No. 114946 - Rectangular cells 10 mm and/or
Cat. No. 114944 - Rectangular cells 50 mm
ater for analysis EMSURE® or
• Cat. No. 116754 - W
distilled water
• Membrane filters, pore size max. 0.45 µm (optional)

33.5 Preparation

• Filter turbid sample solutions over a membrane filter.

33.6 Procedure and measurement

• Open the methods list (<Methods>) and select Method No. 2573 resp. 2574 “Yellowness”.

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• Zeroing the photometer:

• Fill the sample solution into a corresponding rectangular cell and insert the cell into the cell compart-
ment. The measurement is performed automatically.

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A () appears behind the cue “Insert cell” 2 .

• Confirm the measurement by clicking on <OK> 3 .

The measurement results appear in the photometer display 4 .

• Click <START> 5 to start the measurement procedure for the next sample.
A renewed zero adjustment is not necessary.

33.7 Evaluation

Statement of the results: Y

ellowness (YI)
Tristimulus values X, Y, Z
determined for standard illuminant C and the 2° standard observer

33.8 Literature

1. ASTM E313-15, Standard Practice for Calculating Yellowness and Whiteness Indices from
Instrumentally Measured Color Coordinates
2. DIN 6167:1980-01, Description of yellowness of near-white or near-colourless materials

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List of smart icons on display

Main menu Buttons Description

Method list
List of all methods, irrespective of mode

Settings
This button is used to activate method-specific settings
(e.g. sample dilution, turbidity correction, zero adjustment, sample blank, reagent
blank)

Ad hoc
For performing measurements (absorbance/transmission, spectrum, kinetics)
Allows measurements to be performed without the need to create methods

AQA
Overview and list of all Analytical Quality Assurance (AQA) modes

Results list
List of all stored results

System – instrument setup

This button is for optional instrument settings (e. g. date, time, updates etc)

Timer list
List of stopwatch functions

Info Buttons Description

Methods information

Main menu selection button – switches between 2 main menu overviews

Switch between different citations (NH4, NH3 etc)

Switch between different units (mg/l, ppm etc)

Sub-menu Buttons Description

Absorbance/Transmission Mode
Ad hoc submenu: perform absorbance or transmission measurements
Result list: filter concentration mode

Concentration
Method list: create methods -> Concentration Mode
Result list: filter Ad Hoc ABS/Trans measurements

Spectrum Mode
Ad hoc submenu: record spectrum
Method list: create methods -> Spectrum Mode
Result list: filter spectrum mode

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Sub-menu Buttons Description

Kinetic Mode
Ad hoc submenu: perform kinetic measurement
Method list: create methods -> Kinetic Mode
Result list: filter kinetic mode

AQA Status 1&2

AQA submenus: Status display of the period of validity and the outcome (passed /
failed)

AQA1
AQA submenu: List of AQA1 methods

AQA2
AQA submenu: List of AQA2 methods

Pipette check
AQA submenu: List of pipette checking methods

Information
System submenu displays the following information about the device:
Software/method versions, device class, lamp counter and serial number

Interface
System submenu displays the following settings options – and standard settings:
Audible signals – ON, Backlight – 100 %, Print to pdf – ON

Region
System submenu displays the following settings options – and standard settings:
Language, date, time and country zone EU/US, decimal separator – "." (dot)

Quality
System submenu displays the following settings options – and standard settings:
Quick zero – OFF, AQA1 and AQA2 lock – OFF, Zero Adjustment expiry – ON
(interval: 7 days), Use expired reagents – OFF, Service reminder – ON

Automation
System submenu displays the following settings options – and standard settings:
Energy saving mode – ON (10 minutes), Auto Power off – OFF, Auto log off – OFF,
Auto store – ON, Auto print – OFF, Sample ID popup – OFF

User management
System submenu displays the following settings options – and standard settings:
Activation of user management and administrator settings, User login required –
OFF

Service
System submenu displays the following settings options:
Various service functions such as backup, restore, export of log or system data
and import of methods

Update
System submenu displays the option for performing software and method updates

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Sub-menu Buttons Description

Network
This submenu of the “Instrument settings” menu displays the setting options for
connecting the Prove device with a network

Prove Connect
This submenu of the “Instrument settings” menu displays the setting options for
connecting the Prove device with Prove Connect

Selection & Description

Action Icons

Start

Start zero
Start zero adjustment for a method

Apply

Save

Stop

Logout
User logout

Search method

Reset resp. clear filter options

Edit
For editing parameters

Create method

Duplicate/copy
The selected method is duplicated/copied

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Selection & Description

Action Icons

Export button
All selected methods are exported to an external memory device

Import button
Updates/methods are imported from an external memory device into the instrument

Delete
The selected items are deleted

Notification Icons in Description

Menu "Settings"

Dilution
Activate and notify predilution

Turbidity on
Activate and notify the turbidity correction

Show absorbance
Activate and notify display of absorbance value in the result screen

Zero adjustment
Perform zero adjustment

Sample blank value

Activate and notify Sample blank value

Reagent blank
Activate and notify user-defined Reagent blank

Recalibrate
Activate and notify user-defined recalibration

MatrixCheck
Activate MatrixCheck

User defined measurement range

Activate user-defined lower and upper limit of the measuring range

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Toggle Button Description

OFF/ON Button
0 = Off, I = On – the part displayed in light grey is active – here: 0 = OFF

Date/measurement
Switch between date or measurement interval (AQA2); active here: measurement
interval

Absorbance/transmission
Switch between absorbance or transmission mode; active here: transmission mode

Spiking/dilution
Switch between spiking and dilution (MatrixCheck); active here: dilution

Action Icons on Description

Datepicker/Keyboard/
Calculator

Back

Clear

Delete

Apply

Add

Radio Buttons/ Description

Checkboxes

Warning
Warnig symbol check info box

Locked
Change password

Choosen
Check mark

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Selection & Description

Action Icons,
e.g. Result List

Search list
Search function, search criterion: method number, method name or article number
(first 6 digits)

Set date/date filter

Sample ID
Search/results list. Search function, search criterion: sample ID

Select all/select none

Panorama view
Graphic representation of measurement series (control card, control chart, for trend
analyses)

Selection & Action Icons Description

e.g. User-defined Me-
thods

Set value pair(s) for method calibration

Set formula for method calibration

Show graphic view

Selection & Action Icons Description

e.g. Spectrum Kinetic

Table view of values

Return to the recorded spectrum

Next left/Step/next right

Zoom out

Zoom in

View of peak max of a spectrum

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Selection & Action Icons Description

e.g. Spectrum Kinetic

View of peak min of a spectrum

Calculate and show sum of spectra

Calculate and show difference of spectra

Overlay of spectra

First-order derivative of a spectrum

Navigation in graphic view

Status Icons Description

Attention
Warning symbol check info box

Time
Time Stamp

Passed
Status of a check; ü = passed

Off
Status of a check; = inactive

Failed
Status of a check; - = failed

Expired
Status of a check; ! = overdue

Progress
The instrument is in progress

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We provide information and advice to our customers on application technologies and regulatory matters to the best of our knowledge and ability, but
without obligation or liability. Existing laws and regulations are to be observed in all cases by our customers. This also applies in respect to any
rights of third parties. Our information and advice do not relieve our customers of their own responsibility for checking the suitability of our products
for the envisaged purpose.

The life science business of Merck operates as MilliporeSigma in the U.S. and Canada.

Merck KGaA, Frankfurter Straße 250, 64293 Darmstadt, Germany

Merck and Supelco are trademarks of Merck KGaA, Darmstadt, Ger-

many or its affiliates. All other trademarks are the property of their
respective owners. Detailed information on trademarks is available via
publicly accessible resources.
© 2020 Merck KGaA, Darmstadt, Germany and/or its affiliates.
All Rights Reserved.

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Color Prove Manual MK

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Color Prove Manual MK

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Manual

Methods for color

III Typical areas of application . . . . . . . . . . . . . . . . . . . . . . . 10

3 ASTM color measurement . . . . . . . . . . . . . . . . . . . . . . . . 24

4 CIE color distance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

5 CIELAB color space (brightness, chroma) . . . . . . . . . . . . 31

6 CIELUV color space . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34

7 CIExyY color space . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

8 Color (ASBC method) . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

9 Color (EBC method) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47

11 Color 410 acc. to EN 7887 . . . . . . . . . . . . . . . . . . . . . . . . 57

12 Color – Spectral absorption coefficient

13 Gardner color measurement . . . . . . . . . . . . . . . . . . . . . . 68

14 Hess-Ives color scale . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71

15 Hunter color distance . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74

16 HunterLab color space . . . . . . . . . . . . . . . . . . . . . . . . . . . 78

17 ICUMSA Color GS1/3-7 . . . . . . . . . . . . . . . . . . . . . . . . . . 81

18 ICUMSA Color GS2/3-9 . . . . . . . . . . . . . . . . . . . . . . . . . . 85

20 ICUMSA Color GS9/1/2/3-8 . . . . . . . . . . . . . . . . . . . . . . 93

21 Iodine color number . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98

22 Klett color index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103

23 Saybolt color measurement . . . . . . . . . . . . . . . . . . . . . . . 107

24 Spectral absorption coefficient, SAC α(254)* . . . . . . . . . 110

25 Spectral absorption coefficient, SAC α(436) . . . . . . . . . . 114

26 Spectral attenuation coefficient, SAC µ(254)* . . . . . . . . 117

27 Spectral attenuation coefficient, SAC µ(254) -

28 Tint index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125

29 Transmittances TX, TY, TZ . . . . . . . . . . . . . . . . . . . . . . . . . 128

30 UV-absorbing organic matter (UV absorption 254)* . . . . 132

31 UV irradiation (UV absorption 254 -

List of smart icons on display . . . . . . . . . . . .150

* not for Spectroquant® Prove 100

III Typical areas of application

Industry Water Others

III Typical areas of application

1.2 Measuring range

Method 2518 ADMI 50 2.0 - 100.0 ADMI

Method 2517 ADMI 10 10 - 600 ADMI

Method 2518 ADMI 10 10 - 1000 ADMI

1.3 Sample material

Clear liquid samples

1.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

• Filter turbid sample solutions over a membrane filter.

1.6 Procedure and measurement

Zeroing the photometer:

 A () appears behind the cue “Insert cell” 2 .

The measurement result appears in the photometer display 4 .

Statement of the results: ADMI Units

1.8 Method control

1.9 Adjustment (Method 2517 and 2518 only)

• Click <Settings> 6 and select the list “FACTORS” 7 .

2.2 Measuring range

Method 2587 Anisidine value 0.0 - 200.0 AV (Anisidine value)

2.3 Sample material

Animal and vegetable fats and oils

2.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

2.5 Preparing the solutions

p-Anisidine, anhydrous cream-colored crystals:

2.7 Procedure and measurement

• Pre-reaction test solution (A0)

Zeroing the photometer:

Statement of the results: Anisidine value

3 ASTM color measurement

3.2 Measuring range

Method 2562 ASTM Color 0.5 - 8.0 ASTM

3.3 Sample material

3.4 Reagents and auxiliaries

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

3.6 Procedure and measurement

Zeroing the photometer:

A () appears behind the cue “Insert cell” 2 .

The measurement result appears in the photometer display 4 .

Statement of the results: ASTM Units

4 CIE color distance

27 Spectral attenuation coefficient, SAC µ(254) -

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

• Filter turbid sample solutions over a membrane filter.

A () appears behind the cue “Insert cell” 2 .

• Click <Settings> 6 and select the list “FACTORS” 7 .

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

Statement of the results: Anisidine value

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

• Filter turbid sample solutions over a membrane filter.

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

• Filter turbid sample solutions over a membrane filter.

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

• Filter turbid sample solutions over a membrane filter.

• Zeroing the photometer:

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

• Filter turbid sample solutions over a membrane filter.

• Cat. No. 173016 - Spectroquant® VIS Spectrophotometer Prove 100 or

• Degas the sample to remove any carbon dioxide.

• Click on <START> 7 to switch to the measurement procedure.