-: l'

41. Oils and Fats

1
:t

]'- r 'i,

David Firestone, Chapter eaitg1

t.'
Martin Peter Yurawe cz, .Chapfer Editor
U.S. Fpod and Drug, Administrartion' ii
,l

41.1.01 41.1.03 i,!

,,,AOAC Official Method 981.11 AOAC Officiat Method 984.20
Oils and Fats Loss on Dryiiig (Moliture; andiVotatite Matter
,in,Oils and,Fats
,, r Preparation of Test Sample
Procedure Karl Figcher Method
First Action 1981 F"irlf i{atio.h,1984
ri'riit'actian'lsas
Use clear sediment-free liquid sample directly after inverting the .
container several times. If liquid sample contains sediment, release I so/Tcs4ls c1 t Ng*aoac uithoa
4ll gediment from walls of container and distribute uniformly
throughout the oil for deteririination of moisture and volatil.. (Not annt-i{teltg'r{ka1m,::r oxidized oils irnd fats.)
For determinations in which results.might be affecteA Uy porsiUt"
presence of HrO (e.g., iodine value), dry sample as foliows: Add A. Apparatus and Reagenii , '
anhydrous Na2SOa in proportion of.l-2 g per 1 0 g product, and hold .i
(a) Karl Fischer titration' as.1e4nbU.-Memual or automated,
in oven at 50oC. Stir vigorously and filter to obtain clear filtrate.
., Prepare solid sample for determination.of moisfure and volatile
(b) Karl Fischer reagent.-stabilized with HrO equivalent of
matter as directed in 926.12 (s ee.41.1.02). For other determinations,
ca 5 mg HrO/mL reag6nti,,Availa6le.commercially or prepare as
melt test sample in drying oven at a temperature at least I OoC above
mp. If clear, proceed directly. If tubid or contains sediment, filter follows: Dissolve 133 g l, in 425 mL dry pyriding in dry
test sample inside oven. For determinations which might be affected glasq'stoppered,bgttle..
$dd,425 qL,dry ethylene glycol
by the presence ofmoisture'(e.g., iodine value), heat portion oftest monom_ethy! ether. qqol to <49C in ice bath and bubble in I 02- I 05 g
sample in drying oven at 50oC, or l0oC above mp, adding l-2 g SOr. Mix well and let stand 12 h. Reagent is reasonably stable, but
anhydrous Na2SOa per l0 g prciduct. Mix test sample tho.oug:t t! uni restandardize for each series ofdeterrninations. Standardizn daily
filter in drying oven. To retaid rancidity, keep oils and fats in cool with sodium tartrato 2H2O . I mg sodium tartrate .2H2O : 0 .1566 ng
place and protect from light and air. HzO, Alternatively, standardize with'weighed HrO in methyl
Reference.: JAQ AC 64, 429(1981). alcolol as follows: fraiiifer_ iCguratety yeignug.;o*i (:,0. m;
vesse!:and titrate to pteJqometric,end point.
_H;9 to..tirution
Calculate"C: mg I{rOlmL reagent.
..
41.1.02
:

. @),.$p,1!. f i;,cl1ey. reagent .diluentrJ, Methoxyethanol-pyridine

AOAC Officia! Method 926.12 (4+ l).
i
Moisfure and Volatile Matter (d) Test sample solvent.-Anhydrous CHClr-methyl alcohol (l
in Oils and Fats : +lor2+l). ;;' r :,: ilrlii
:" ' ' r . : ] :!'
Vacuum Oven Method ,

[iirl'l]t
' Flrst Action 1926 i
Bt, Determinqligq: :, .,,,..
Final Action
",[ei8fr,r1g,ne?rest 9,9i,8, 5-25 g pre,pared test portion,
Softert sample, if necessary, by gentle heat, taking care not to melt it. centaining <109:.@g, tlror,,into tiqation vesqel, dissolve in
When soft enough, mix thoroughly with effective mechanical-mixer anbyr,flpuu, CHCl3--methyl .alcohol. Titrate. w,ith.undiluted or
Weigti S t O.Z g prepared test portion into Al moisture dish ca 5 cm ililuted (1, + l),Karl'lFischer reagent to electrornetric end point:
diameter and 2 cm
deep with tight-fit slip-over cover. Dry to Car,ry'outlb.lank te$t Using Same amouots of reagent, diluenl and
solvents. Subtiact blank' titer.
constant weight in vacuum oven at uniform temperature 20"*25oC
abovebp of HrO atworkingpiessure, which shouldbe <100 mmHg
(13.3 kPa)..Cool in efficient desiccator 30 min and weigh. Constant mL reagent xC
H.O.yo_
weight is attained when successive I g test portion xI0
h drying periods show
additional loss of <0.05%. Report % loss in weight as moisture and
volatile matter. (Nofe: Pyridine-free Karl Fischer reagents are available from
laboratory reagent suppliers.)
Rbferences: Ind. Eng. Chem. 18, 1347 (1926).
AOAC OFFTC|AL METHODS oF ANALYS|S (2016) OILS AND FAT
Chapter4l, p.3

If test sample and HrO are weighed in same pycnometer, at same firmly and move alidade backward or forward until field of vision is
temperarure (7), divided into light and dark portion. Line dividing these portions is
"border line," and, as a rule, will not be sharp line but band of color.
Specific gravity, = W1,,1" Colors are eliminated by rotating screw head of compensator until
^r1!1tr11,o
sharp, colorless line is obtained. Adjust border line so that it falls on
Repeatability.-Difference between results of 2 determinations
point of intersection of cross hairs. Read n of substance directly on
done simultaneously or in quick succession by same analyst should
scale of sectoq estimating 4th decimal place. Take )3 readings,
not exceed 2 units of 4th decimal place.
approaching intersection alternately from orle field to other, and
References: Standard Methods for the Analysis of Oils, Fats, and
Deriyatives (1979) 6th Ed., Pergamon Press, New
York, NY, USA.
Taylor, J.K. (1967) Treatise on Analytical Chemistry, Table 921.08. Butyrorefractometer readings and indexes of
refraction
I.M. Kolthoff & P.J. Elving (Eds), Interscience
Publishers, New York, NV, USa, Part I, Sec..D-4, lndex of lndex of
Vol. 7, Ch. 8l, "Measurement of Density and refraction
Specific Gravity,': pp 4561-4610. 40.0 1.4524 60.0 '1.4659

40.5 1.4527 60.5 1.4662

41.0 1.4531 61.0 1.4665
41.1.07 '1.4668
41 .5 1.4534 61.5
AOAC Official Method 921.08
. lndex of Refraction
42.0 1.4538 62.0 1.4672

of Oils and Fats 42.5 1.4541 62.5 1.4675

43.0 1.4545 63.0 1.4678
43.5 1.4548 63.5 1.4681
44.0 1.4552 64.0 '1.4685
A. General Directions
44.5 1.4555 64.5 1.4688
Determine index of refraction (n) with any standard instrument,
45.0 1.4558 65.0 1.4691
reading oils at 20" or 25oC and fats at 40oC. Place instrument so that
45.5 1.4562 65.s 1.4694
diffi.rsed daylight or some form of artificial light such as Na vapor
lamp can be used for illumination. Circulate stream of constant 46.0 1.4565 66.0 1.4697
temperature (10.2"C) HrO through prisms. Approximate 46.5 1.4569 66.5 1.4700
temperature corrections of bufyrorefractometer readings may be 47.0 1.4572 67.0 1.4704
made by following formula: 47.5 1.4576 67.5 1.4707
48.0 1.4579 68.0 1.4710
]?:R,+K(7,_I) 48.5 1.4583 68.5 1.4713

where R: reading reduced to standard temperature, R' reading : 49.0 1.4586 69.0 1.4717
obtained at temperature I', I:
standard temperature, and, K: 49.5 1.4590 69.5 1.4720
0.55 for fats and 0.58 for oils. 50.0 1.4593 70.0 1.4723
Readings of instruments that give n directly can be reduced to 50.5 1.4596 70.5 1.4726
standard temperature by substituting factor 0.000365 for 0.55 and 51 .0 1.4600 71.0 1.4729
0.000385 for 0.58 in formula. As temperature rises, n falls. 5'1.5 '1.4603 71.5 1.4732
Instrument used may be standardized with H2O at20"C, theoretical
52.0 1.4607 72.0 1.4735
n of H2O at that temperature being 1.3330. Any correction found
52.5 .4610 72.5 1.4738
should be made on all readings. .Indei of refraction varies with
1

densiry and in saine direction. 53.0 1 .4613 73.0 1.4741

53.5 1 .4616 73.5 1.4744
B. By Abbd Refractometer
54.0 1 .4619 74.0 1.4747
To charge instrument, open doubleprismby means of screwhead
54.5 1.4623 74.5 1.4750
and place few drops test sample on prism or, if preferred, open
55.0 1.4626 75.0 1.4753
prisms slightly by tuming screw head ard pour few drops test
sample into funnel-shape aperture between prisms. Close frisms 55.5 1.4629 75.5 1.4756
firmly by tightening screw head. Let instrument stand few min 56.0 1.4633 76.0 1.4759
before reading, so that temperature of test sample and instrument 56.5 1.4636 76.5 1.4762
will be same. Clean prisms between readings by wiping offoil with 57.0 1.4639 77.0 1.4765
soft cloth, then with cotton pad moistened with solvent (e.g., 57.5 '1.4642 77.5 1.4768
trichloroethylene, toluene, orpetroleum ether), and let dry. 58.0 1.4646 78.0 1.4771
Method of measurement is based upon observation of position of
58.5 1.4649 78.5 1.4774
border line of total reflection in relation to faces of flint glass prism.
59.0 1.4652 79.0 1.4777
Brins this border line into field of vision of telescooe bv rotatins
 -.- .:i.ii11 (:..'j::.,. :i.; i:

, '?,,' ,,i:,
41.1.'t6
AOAC Offiqiat Method 965.33
Peroxide Value of Oils and Fats
.l
Titration Method
First Action 196e ' ::
jli i' Fiiiil.AGtio#*069;' ;.,

AOC*AOAC Method

(iy'ore: Conduct analysis in difiiise daylight or in artificial light

shielded from direct light source.)
A. Reagents
(t) Acetic acid-chlorbform solution.-Mix 3 volumes
CH3COOH with 2 volumes CHCI3, USP.
(b) Potassium iodide solution, saturated.-Dissolve excess KI
in freshly boiled H2O. Excess solid must remain. Store in dark. Test
daily by adding 0.5 ml to 30 mL CHTCOOH-CHC1r, (a); then add
2 drops.l%o starch solution, (mix ca 1 g soluble starch with enough
cold rpater to make thin paste, add 100 mL boiling HrO, and boil ca
I min while stirring). If solution turns,blue, requiiing >1 drop 0.1Iv{
Na2:S2O3 to discharge color, prepare fresh solution.
(c) Sodium thiosulfate standard solutions.-4.1 and 0.0iM.
Prepareand-standardize as in 942.27 (see A.1.73). For 0.011v1, dilute
0.1M with freshly boiled and cooled HrO.

i
OILS AND FAT
\ Chapter 41, P.12

B. Determination
5 00 i 0 05 c itto 250 mL
(a) Fats and olls.-Weigh tT l"^1:l
(a)'
glass-stoppered Erleruneyer' Add 30 mL CHTCOOH-CHC1:'
0.5 mL saftirated KI solution' (b)' from
olrrO s*lri io dissolve. Add
add 30 ml
frlot, plp",, 1et stand with occasional shaking 1 min' and
shaking until
HrOr Sl"*fy titrate with 0.1M Na2S2O3 with vigorous
solution' and
yato* is aimost gone. Add ca O'S rnl- 1% starch
to release all from CHC13
Lontinue titration, ,fruki,g vigorously
12
is used'
iuy.., untit blue just disappears' If <0'5 mL 0'1M Na2S203
repeat determination with 0'01M Na2S2O3'
(must be <0'1 mL 0'1M
Clonduct blank determination daily
Na2S2O3). Subtract from test portion tihation'

Peroxide value (milliequivalent peroxidelkg

oil or fat): S x M x
= mL NarSrO, Qlaok correcte d) and M
1000/9 test portion, where .S

: molarity NazS,O3 solution'

fat by heating with constant stirring on
hot
(b) Margarine.-Melt
(Avoid excessive
plate at lo,Jh.ut, or heat in air oven at 60"-70oC'
melted' hold in
heat and long exposure >40"C') When completely-
most have settled'
warm place until aqueous portion and
9f -:-tidt
beaker and filtqr through WhatrnanNo' 4' or
Decani oil into clean
paper. Do not reheat unless necessary to obtain clear
equivalent
filtrate. Proceed as in (a).

References: J. Am. Oil Chem' Soc' 26,345(1949)'

AOCS Method Cd 8-53'
JAOAC 48,17s(1965)'

!. :- r-
 41.1.21
' AOAC Officiat Method g4l.2g
. -Fatty Acids (Free)
in Crude and Reiined Oits
Titration Method !,.i

First Action 1940

Final Action

N ationat Cottonseed prod ucts Associati

ot>AOAC Method
(a) In crude oils.-weigh
7.05 g well-mixed oil into 250
mL
flask or4 oz bottle. Add 50iriri;;;i,;;.d"r;;##.H
addr.ns 2
yrnhenolphthalei"
prbduce faintpermanentpink.
r"tr;;;;;;lirn 0.,, *uor,o
Itrate wlttr o.ilii r,r"oa,
(see A.t.t2), with vigorous ,h.k;;;;;;l i.'#o.o, faint%6.16
pin_k
appears and persists
Report as percent free fafy acids
=,
expressed as oleic acid. Tjr.
(Using,il. ,"_pf" ,ir" and molarity of
),:"o*r::f.d in the methodjhe
-il:r; ,iuo, u,.o in the _
sample.) ,"ffi",f ii:
(b)
In refined oils.-Jo ca 50 mL
alcohol in clean, dry 150 mI
flask, add few drops of the
in HrO
oil ano zJpi.*tptffi; piace flask
at 60.-65"C until

l,:""'-':iTi'-;#il,#T;,X11,?Y;n';*.';'IJf
alcohol and tihate with
0
,1J;
occasionary*.,r^,"r""ilf"I,f#:i*,'JfitrJ:r;:ri;:A;

AOAC OFF|CIAL METHODS OFANALysts (2016)

fal1t f3nnanent pink appears in supernate alcohol. Multiply

mL
0.1 M NaOH by 0.05 and report
as percent free fatty acids expressed
as oleic acid.
Free fatly acids may also be expressed in terms
of acid value
(mg KOH necessary to neutralize 1 g oil).

Acid value : percent free fatg acids x 1.99

1as oleic.)

Revised: April 2012