Exercise I: Assessment of Accuracy and Precision • Should be used when accuracy and reproducibility

are crucial, because these can achieve accuracy to

Principle of Method four significant figures.
Proper use of equipment and glassware in analytical tests • When emptying a volumetric pipette, the liquid is
helps ensure more accurate and precise results. allowed to drain out. It is NOT forced out.

Note: Graduated pipette:

Each piece of volumetric glassware is marked with its total • Calibrated into small divisions so that various
volume, the notation TD or TC, and a temperature (usually amounts of liquid can be measured with the same
20°C). pipette.
• Usually used to measure any amount between 0.1ml
and 25.0ml.
• They are not as accurate due to the fact that any
imperfection in their internal diameter will have a
greater effect on the volume delivered.
• After all the liquid has been allowed to drain out by
Printed on each piece of volumetric glassware are the
gravity, the last drop remaining in the tip is also
specifications that indicate:
forced out.
• the maximum volume of liquid that can be
Calibrated pipettes are divided into:
transferred
• MOHR PIPETTES
• the size of the divisions on the pipette
– the graduations on these always end before
• the temperature at which calibrations were made
the tip
 Since density and volume change with
temperature, the volume markings are
strictly correct only at the calibration Mohr Pipette
temperature. • SEROLOGICAL PIPETTES
• if the pipette is a “to deliver”(TD) or “to contain” – the graduation marks continue to the tip
(TC) pipette.
 TD is used on burettes and pipets (and
some graduated cylinders), means that the Serological Pipette
apparatus is calibrated to accurately deliver
or transfer the stated volume to another (2) Difference between markings on a 10-ml versus a
container. 25- or 50-ml buret.
 TC is used on volumetric flasks and (most)
graduated cylinders, means that the Most popular burettes are 10 mL, 25 mL and 50 mL types.
markings give an accurate measure of the 10 mL burettes are usually graduated each 0.05 mL, while
volume contained, but that pouring the 25 mL and 50 mL burettes are usually graduated each 0.1 mL.
liquid into another container will not
necessarily deliver the indicated volume. Table 1. Relative accuracies of volumetric glasswares
mL pipette burette flask
(1) Difference between dispensing from a volumetric tolerance relative tolerance relative tolerance relative
pipette and a graduated pipette mL (%) mL (%) mL (%)
1 0.006 0.60 0.010 1.00
2 0.006 0.30 0.015 0.75
Volumetric pipette:
3 0.010 0.33 0.015 0.50
• Used to deliver a single specific volume of liquid,
4 0.010 0.25
usually between 1 and 100 ml.
5 0.010 0.20 0.020 0.40
• Shaped like rolling pins with a large belly, one blunt 10 0.020 0.20 0.020 0.25 0.020 0.20
end, the neck, and one tapering end, the tip. 15 0.030 0.20 15
20 0.030 0.15 20
25 0.030 0.12 0.030 0.15 0.030 0.12
50 0.050 0.10 0.050 0.13 0.050 0.10
Volumetric pipette
• Used for accurate measurements, since it is designed • The lower the volume of glassware, the higher the
to deliver only one volume and is calibrated at that relative error. Thus, for high precision work we
volume. should use glassware of higher volume.

FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 1
A procedure or measurement technique is validated by
generating numbers that estimate their accuracy and
precision.
* If the number of replicate determinations is ≥30
Accuracy is a measure of how close a measured value is to
the accepted or “true” value.
• It is related to the percent error between the
average volume of solution measured Example 1:
experimentally and the volume that was expected Table 1. Determination of the Standard Deviation of %
(the accepted value). Moisture in Uncooked Hamburger
• Accuracy is sometimes estimated by the relative
error (%Erel) between the mean analysis value and
the true value.
• Smaller %Erel reflects higher accuracy.
• Percent error can be negative, indicating that the
measured volume was smaller than the expected
volume or positive, indicating that the measured
volume was larger than the expected volume.
• Absolute precision is reflected by the standard
deviation (SD), while relative precision is calculated
as the coefficient of variation (CV).

Precision measures the closeness of a set of values obtained

from identical measurements of the same quantity.
• It is the measure of reproducibility of a
Coefficient of variation (CV) or relative standard deviation
measurement (whether it’s accurate or not).
• Precision is often called error. represents the ratio of the standard deviation to the mean.
• Precision is related to the standard deviation of a
series of measurements of the same thing.
• For example, if the micropipet is set to the same
volume (300 μL) and four measurements are taken
at this volume, a standard deviation can be taken of
these five measurements; the smaller the standard *Low CV indicates a high level of precision or reproducibility
deviation, the more precise the micropipet is. of the replicates. As a rule, a CV below 5% is considered
acceptable.
Range is the difference between the largest and smallest
observation. *For large numbers of samples, we can determine the
Standard deviation (SD or σ) measures the spread of the confidence limit or interval around the mean using the
experimental values and gives an indication of how close the statistical parameter called the Z value.
values are to each other.  We do this calculation by first looking up the Z value
Formulas of SD: from statistical tables once we have decided the
desired degree of certainty.
Table 2. Values for Z for Checking Both Upper and Lower
Levels

 If n= ≥ 25 use this formula:

*If the number of replicate determinations is small (≤30)

FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 2
 Using the data for the percent moisture of uncooked
hamburger, suppose the true value of the sample is 65.05%.
CI based on Example 1 given n= 25:
The percent relative error is calculated using our mean value
of 64.72%

 If n is small look up the t value table based on the

degrees of freedom, which is the sample size minus
one (n − 1), and the desired level of confidence to *Note that we keep the negative value, which indicates the
compute for confidence limit or interval direction of our error, that is, our results were
0.507% lower than the true value.
Table 3. Values of t for Various Levels of Probability
Table 4. Viscosity and Density of Water at Various
Temperatures

SELF TEST
1. How are standard deviation, coefficient of variation,
mean, % relative error, and 95% confidence interval
The calculation for Example 1 with four samples (n) and three
affected by: 1) more replicates, 2) larger size of
degrees of freedom (n−1) is given below:
measurement?
 Generally, increasing the size of measurement (n)
has a direct effect on the SD of the replicates given
that there is precision. Specifically, the SD of the
replicates reduces by a factor of the square root of n.
For example, all other things being equal, if the SD of
replicates is 4.0 with n = 30, doubling the increments
*Another way to determine accuracy is to compare the to n = 60 would reduce SD by the square root of 2
results with an existing true value. In this case, we can (or 1.414) to approximately 2.8.
calculate the error for our test, compare it to the known  Increasing the size of measurement (n) reduces the
value, and determine the accuracy. potential for errors in terms of both frequency and
degree of underestimation of the mean.
Absolute error is the difference between the experimental  THUS, THE BIGGER THE SAMPLE SIZE, THE MORE
value and the true value. CHANCES OF GETTING AN ACCURATE DATA
 The absolute error term can have either a positive or BECAUSE THE MORE CONFIDENT IT GET, THE
a negative value. If the experimentally determined SMALLER THE ERROR WILL BE
value is from several replicates, then the mean (0)  Increasing replicates (r) does not reduce the SD but
would be substituted for the x term. it does improve the estimate of the SD by reducing
the variability in the estimate. Likewise, increasing r
(%) Relative error results are reported as a negative or reduces the standard error for the grand mean.
positive value, which represents a fraction of the true value. Specifically, the standard error reduces by the
 If desired, the relative error can be expressed as square root of r.
percent relative error by multiplying by 100%.  THUS, THE MORE THE REPLICATES THE LESSER THE
ERROR WILL BE

FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 3
  VARIANCE (DIFFERENCE FROM THE MEAN) AND SD
IS DIRECTLY PROPORTIONAL
2. Why are % relative error and coefficient of variation used
to compare the accuracy and precision, respectively, of
the volumes from pipetting/dispensing 5 and 10 ml with
pipette and burette rather than simply the mean and
standard deviation respectively.
o If there are several experimental values,
 % relative error and CV is more appropriate to use in
then the relative average deviation from
determining the accuracy of and precision of the
the mean becomes a useful indicator of
glassware since SD is heavily influenced by outliers
precision. It is calculated similarly to the
just like the mean. In addition, if all values are the
relative deviation from the mean, except
same, then the standard deviation is 0. However, the
the average deviation is used instead of the
CV helps us see that even a lower SD doesn't mean
individual deviation.
less variable data. While % relative error specifies
whether the measurements were done in excess or
deficient.

3. Would you expect your accuracy and precision to be

better using a 25-ml buret or a 50-ml buret? Why? Exercise 2: Determination of Moisture
 Accuracy and precision would be better for a 50- ml
burette because it has the highest resolution and Principle of Method
lowest % relative error in comparison to 10-ml and The sample is heated under specified conditions and the loss
25-ml burette. of weight is used to calculate the moisture content of the
o 0.050 mL out of 50 mL is 0.1%, and that's sample.
about maximum precision that we can get
from volume measurement when using
burette. In turn that's also about the
maximum precision of the titration.

4. You are considering adopting a new analytical method in

your lab to measure moisture content of cereal products.
How would you determine the precision of the new
method and compare it to the old method? Include any
equations to be used for any needed calculations.

 The best and most commonly used statistical

evaluation of the precision of analytical data is the
SD. The standard deviation measures the spread of
the experimental data and gives a good indication of
how close the new method is to the old method. In
getting the SD, we must first compute for the mean.
Once we have a mean and standard deviation, we
next must determine how to interpret these
numbers by getting the coefficient of variation. As a Example 1
rule, a CV below 5% is considered acceptable. Carrots Replicate I Replicate II
o If the sample is large we can determine the Pan (g) 45.48g 46.05g
confidence interval using the Z value to Pan + Wet sample 50.51g 51.09g
establish the true mean value. (g)
 Other quick tests of precision used are the relative Pan + Dried sample 45.97g 47.29g
deviation from the mean and the relative average (g)
deviation from the mean. % Moisture
o The relative deviation from the mean is
useful when only two replicates have been
performed
FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 4
 %MC= 100- 85= 15%

5. Give the function of the following: drying oven,

desiccator, tong, analytical balance, muffle furnace
A. Drying oven- used for drying samples,
glassware and crucibles to constant
weight. Gravity convection ovens circulate
air in the oven through holes at the top and
SELF TEST
bottom of the oven. This circulation of air
1. What is the economic importance of moisture
keeps the temperature even throughout
content of foods?
the oven and keeps the humidity low.
 Moisture content is of great economic
B. Muffle furnace- is a kiln type oven, capable
importance to a food manufacturer because
water is an inexpensive filler. Moisture is also of maintaining extremely high temperatures
an important factor in food quality, (up to 1000◦C) for extended periods of
preservation, and resistance to deterioration. time. Muffle furnaces are used primarily for
Determination of moisture content also is igniting organic solids in the determination
necessary to calculate the content of other food of volatile solids.
constituents on a uniform basis. C. Dessicators- are containers which are used
 Importance of determining moisture content
to provide a moisture-free environment for
includes:
o Legal and labelling requirements cooling and storing objects and
o Microbial stability chemicals. The top and bottom fit together
o Food quality to form an airtight seal. The bottom section
o Food processing operations of the desiccator contains a chemical
(desiccant) which absorbs moisture from
2. Arrange the following samples according to
the air.
increasing shelf-life: Sweet potato tops, Carrots, D. Tongs- are special devices for grasping
Pineapple, Fish, Cake flour.
glassware. It is very important to use tongs
Sample % MC Shelf- life for handling crucibles to keep skin oil off
Cake flour 14
the sides and to prevent burns.
Fish 79.1
E. Analytical balance- is an extremely accurate
Sweet potato tops 82
Pineapple 87 piece of equipment used for determining
Carrots 87 weights of various items. An analytical
balance used for the total suspended solids
3. What method is best to use in determining the test must be capable of weighing to
moisture content of spices? a) Thermal drying by 0.0001g.
ordinary drying oven, b) Thermal drying by vacuum
oven, c) Distillation
 Distillation (Dean and Stark method) is suitable for 6. Hot air oven was used to determine moisture
determining the moisture content of foods with low content; in each case specified would you likely
moisture content. Given that spices contain several overestimate or underestimate the moisture content
volatile compounds, distillation is ideal because its of a food product being tested? Explain your answer.
volatile oils will remain dissolved in the organic  Selections of a representative sample, and
solvent (xylene or toluene) and thus not interfering prevention of changes in the properties of the
sample prior to analysis, are two major potential
with the measurement of water.
sources of error in any food analysis procedure.
 Determination of moisture content (including
4. A bread roll weighed 100g at time of receipt. The overestimation and underestimation) may be
weight after drying was 76.47g. The % solids after affected by the following:
oven drying was 85%. Determine moisture content.
FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 5
 1. Clumping and surface crust formation. Some the food sample during drying, carrying other food
samples tend to clump together or form a semi- constituents with it.
permeable surface crust during the drying o For example, most of the moisture in milk is
procedure. This can lead to erroneous and removed by heating on a steam bath prior
irreproducible results because the loss of moisture is to completing the drying in an oven.
restricted by the clumps or crust. For this reason 8. Temperature and power level variations. Most
samples are often mixed with dried sand to prevent evaporation methods require a definite temperature
clumping and surface crust formation. or power level to dry the sample so as to standardize
2. Elevation of boiling point. Under normal laboratory the procedure and obtain reproducible results. The
conditions pure water boils at 100 oC. But, if solutes efficiency of the drying procedure depends on the
are present in a sample the boiling point of water is precise location of the sample within the instrument.
elevated. Consequently, the rate of moisture loss It is therefore important to carefully design and
from the sample is slower than expected. operate analytical instruments so as to minimize
3. Decomposition of other food components. If the these temperature or power level variations.
temperature of drying is too high, or the drying is 9. Sample pans. It is important to use appropriate pans
carried out for too long, there may be to contain samples, and to handle them correctly,
decomposition of some of the heat-sensitive when carrying out a moisture content analysis.
components in the food. This will cause a change in Typically aluminum pans are used because they are
the mass of the food matrix and lead to errors in the relatively cheap and have a high thermal
moisture content determination. It is therefore conductivity. These pans usually have lids to prevent
normally necessary to use a compromise time and spattering of the sample, which would lead to
temperature, which are sufficient to remove most of weight loss and therefore erroneous results. Pans
the moisture, but not too long to cause significant should be handled with tongs because fingerprints
thermal decomposition of the food matrix. can contribute to the mass of a sample. Pans should
o One example of decomposition that interferes be dried in an oven and stored in a descicator prior
with moisture content determinations is that of to use to ensure that no residual moisture is
carbohydrates. The water that is released by this attached to them.
reaction is not the water we are trying to
measure and would lead to an overestimation of Exercise 3: Titratable Acidity of Fruit Juices
the true moisture content.
o On the other hand, a number of chemical Principle of Method
reactions that occur at elevated temperatures The volume of a standard base used to titrate the organic
lead to water absorption, e.g., sucrose acids in foods to a phenolphthalein endpoint can be used to
hydrolysis, and therefore lead to an determine the titratable acidity.
underestimation of the true moisture content.
6. Volatilization of other food components. It is often The titratable acidity measures the total acid concentration in
assumed that the weight loss of a food upon heating a food. Food acids are usually organic acids, with citric, malic,
is entirely due to evaporation of the water. In lactic, tartaric, and acetic acids being the most common.
practice, foods often contain other volatile  This quantity is determined by exhaustive titration of
constituents that can also be lost during intrinsic acids with a standard base.
heating, e.g., flavors or odors.  Titratable acidity is a better predictor of acid’s
o For most foods, these volatiles only make impact on flavor than pH.
up a very small proportion and can  The organic acids present in foods influence the
therefore be ignored. flavor (i.e., tartness), color (though their impact on
o For foods that do contain significant anthocyanin and other pH-influenced pigments),
amounts of volatile components (e.g. spices microbial stability (via inherent pH-sensitive
and herbs) it is necessary to use alternative characteristics of organisms), and keeping quality
methods to determine their moisture (arising from varying chemical sensitivities of food
content, e.g., distillation, chemical or components to pH).
physical methods.  The titratable acidity of fruits is used, along with
7. High moisture samples. Food samples that have high sugar content, as an indicator of maturity.
moisture contents are usually dried in two stages to
prevent "spattering" of the sample, and Food acids are usually reported as percent of total sample
accumulation of moisture in the oven. Spattering is weight:
the process whereby some of the water jumps out of

FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 6
 most common indicator for food use. It changes
from clear to red in the pH region 8.0–9.6. Significant
Acid Factor color change is usually present by pH 8.2. This pH is
Citric 6.4 termed the phenolphthalein endpoint
Mallic 6.7  Titratable acidity is the number of protons
Tartaric 7.5 recovered during a titration with a strong base to a
Lactic 9.0 specified endpoint. It can also be expressed as a
molar quantity. pH is the logarithm of the
concentration of free protons, expressed with a
Titratable Acidity of Fruit Juice positive sign. There is no direct relationship between
titratable acidity and pH in juice, although generally
Predominant acid

Predominant acid
TA (g/1ooml)

TA (g/1ooml)
the pH goes up as the acid goes down and vice-
Juice

Juice versa. The former measures the total acidity but

does not measure the strength of the acids. The pH
indicates the strength of the acid condition.
Apple 0.36–0.80 Malic Guava 1.16 Citric
Cherry, Sour 0.94–1.64 Malic Lemon 4.0–6.2 Citric
3. Draw the titration assembly.
Cherry, Sweet 0.24–0.94 Citric Lime 7.4-7.8 Citric
Cranberry 1.6–3.6 Citric Nectarine 0.24–0.94 Citric
Grape 0.4–0.9 Tartaric Orange 0.8–1.4 Citric
Grapefruit 1.2–2.0 Citric Mango 0.34–0.84 Citric
Peach 0.24–0.94 Citric Pineapple 0.7–1.6 Citric
Pear 0.36–0.80 Malic Plum 0.94–1.64 Malic
Strawberry 0.6–1.1 Citric Tomato 0.34–1.00 Citric

SELF TEST
1. Give the equivalent weight of a) malic acid b) tartaric
acid c) citric acid
Molecular and Equivalent Weights of Common Food Acids 4. A 10 ml orange juice with a density of 1.09 g/ml
Acid Molecular Equivalents Equivalent required 4.1 ml of 0.1N NaOH for neutralization.
Weight per Mole Weight Determine % citric acid.
Citric 192.12 3 64.04
(anhydrous) D= m/v  m= Dv
Acetic 60.06 1 60.05
Lactic 90.08 1 90.08 m= 10.9 g
Malic 134.09 2 67.05
Oxalic 90.04 2 45.02
Tartaric 150.09 2 75.05
Ascorbic 176.12 2 88.06
Hydrochloric 36.47 1 36.47
Sulfuric 98.08 2 49.04
Phosphoric 98.00 3 32.67
Potassium 204.22 1 204.22 % citric acid= 0. 2407 %
acid
phthalate 5. Forty four ml of NaOH solution neutralizes 1.0253g
of potassium acid phthalate. What is the normality
2. Differentiate the ffg.: Stoichiometric point and end
of NaOH?
point, titratable acidity and pH
 At the equivalence point, or stoichiometric point, in
a titration, the number of acid equivalents exactly
equals the number of base equivalents, and total
acid neutralization is achieved. Whereas, when
N NaOH= 0.1141 N
indicators are used, the term endpoint or
colorimetric endpoint is substituted for equivalence
point. This emphasizes that the resulting values are Exercise 4: Acidity Measurement in Dairy Products
approximate and dependent on the specific indicator
resulting to a color change. Phenolphthalein is the

FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 7
Titratable acidity provides a simple estimate of the total acid
content of a food. In most cases, it is only an estimate since 1) The quality indicator of dairy product is acidity.
foods often contain many acids that cannot be differentiated 2) Phenolphthalein indicator changes its color from
through titration. The predominant acid in dairy products is
colorless to faint pink at pH 8.3.
lactic acid.
3) The process of determining the concentration of
unknown solution by reaction with a solution of known
composition is titration.
4) Titratable acidity is the total potential acidity and it
SELF TEST represents both the free H+ and that which can be
1. What is the significance of determining the derived from the undissociated molecule during titration.
acidity of milk? 5) Total titratable acidity of milk is generally expressed as %
lactic acid.
 This parameter is tested to determine the quality of
the milk product, given that the acidity of milk is Exercise 6: Determination of Fat (Roese-Gottlieb)
relatively constant, hence any increase in acidity
directly affects its taste and overall quality. Fat is an important food component due to its nutritional
 As milk acidity increases over time, measuring this value and economical aspects.
parameter is also a means of monitoring storage  Good energy source essential fatty acids
conditions. Acidity is determined by an end-point  Sensory properties: taste, flavor, consistency and
titration using sodium hydroxide (a base) and is texture of foods
defined as the consumption of base necessary to  Low cost ingredient
shift the pH value from 6.6 +0.1 (corresponding to  Bulk substance
fresh milk) to a determined basic pH value. While pH
7.0 is the actual point of neutralization, Crude Fat is defined as the substance that can be extracted
phenolphthalein is commonly employed as a color by an organic solvent, from an analytical point of view.
indicator to determine the end-point of reaction and
with it, color change occurs at pH 8.3. Fat content is determined often by (a) solvent extraction
methods (e.g., Soxhlet, Goldfish, Mojonnier), but it also can
2. Give the balanced chemical equation of be determined by (b) nonsolvent wet extraction methods
conversion of lactose in milk to lactic acid by (e.g., Babcock, Gerber), and by (c) instrumental methods that
fermentation. rely on the physical and chemical properties of lipids (e.g.,
C12H22O11 + H2O  4 H3CHOHCOOH infrared, density, X-ray absorption).
Lactose + Water Lactic acid
Factors affecting the choice of method:
o nature of the sample (e.g., dry versus moist)
1. Calculate %lactic acid in a sample of butter o purpose of the analysis (e.g., official nutrition labeling
from the ffg. data: or rapid quality control)
Sample = 20g 0.02M NaOH = 2.8 ml o instrumentation available (e.g., simple glassware vs.
expensive instrument)

Factors influencing the choice of solvent:

2. Give the function of the ffg:
o official acceptance
1) phenolphthalein- color indicator o dissolving properties
2) stirring rod- used for mixing; ensures that the titer o polarity
and the titrant are homogenized throughout titration o toxicity
3) pipette- used to transfer and deliver small amounts of o inflammability
the sample o partial vapor pressure
4) burette- analytical dispensing variable that contains o cost
the titrant
Rose- Gottlieb is a gravimetric extraction method which is
5) conical flask- vessel that contains the sample widely used for dairy products.

3. Fill in the blanks to complete the statement: Principle of Method

FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 8
The fat content is determined gravimetrically after extraction 2) Which food from the breads, cereals, rice and pasta
with diethyl ether and petroleum ether from an ammoniacal group generally contain fat?
solution of the sample. A. Biscuits
B. Muffins
C. Pasta
D.A and B
SELF TEST 3) Which vitamin is soluble in fat?
1. What is crude fat? A. Ascorbic acid
 Crude fat is the term used to refer to the rough B. Vitamin A
mixture of fat-soluble material present in a sample. C. Thiamine
Crude fat also known as the ether extract or the free D.Riboflavin
lipid content, is the traditional measure of fat in food 4) Solvent extraction is generally required in food analysis
products. as:
A. Clean- up procedure
Crude fat, which includes phospholipids and wax
esters, as well as minor amounts of non-fatty B. Concentration step
material (AOAC, 2000). Total fat can be expressed as C. To remove slightly soluble material
triglyceride equivalents determined as the sum of D.All of the above
individual fatty acids and expressed as triglycerides 5) Which extraction solvent is best for fat analysis?
(FAO, 1994) A. Diethyl ether
B. Petroleum ether
2. What is the best method of determining fat for foods high
C. Alcohol
in protein and sugar?
D.Hexane
 The Roese-Gottlieb and Mojonnier methods are
often used for the extraction of fat from products
high in protein because it dissolves and precipitates Exercise 7: Determination of Free Fatty Acids
protein in ethanol and ammonia, whereas fats are
dissolved in petroleum ether and diethyl ether. Deterioration of used frying oils and fats can be monitored by
measuring the polar components, which include
3. What is the role of NaCl solution in fat analysis by Rose- monoacylglycerols, diacylglycerols, FFAs, and oxidation
Gotlieb method? products formed during heating of foodstuffs.
 Milk contains emulsions, colloids, molecular and
 Lipid oxidation (also called autoxidation) as it occurs in bulk
ionic solutions. The dissolved salt (NaCl) breaks up fats and oils proceeds via a self- sustaining free radical
into ions which float freely in the solvent and it mechanism that produces hydroperoxides (initial or
becomes an ionic solution of free sodium and primary products) that undergo scission to form various
chloride ions, which can facilitate the precipitation of secondary products including aldehydes, ketones, organic
sodium salts in long chain fatty acids. acids, and hydrocarbons (final or secondary products)

 Rancidity refers to the off odors and flavors resulting from

4. A 0.2 g fat was extracted from 20 g sample of evaporated
lipolysis (hydrolytic rancidity) or lipid oxidation (oxidative
milk. What is the fat content of the sample? rancidity).

 Lipolysis is the hydrolysis of fatty acids from the glyceride

molecule. Because of their volatility, hydrolysis of short-
5. Identify the best correct answer. chain fatty acids can result in off odors.
1) Which of the following is not one of the ways fat are o Fatty acids shorter than C12 (lauric acid)
useful in foods? can produce off-odors in foods.
A. Fats contribute to the taste and smell of foods
FFA is the percentage by weight of a specified fatty acid (e.g.,
B. Fats carry fat- soluble vitamins
percent oleic acid).
C. Fats provide a low- calorie source of energy compared to
carbohydrates Principle of Method
D.Fats provide essential fatty acids

FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 9
Measures of fat acidity normally reflect the amount of fatty
acids hydrolyzed from triacylglycerols. To a liquid fat sample, 2. What is antioxidant? Why are they necessary?
neutralized 95% ethanol and phenolphthalein indicator are  Antioxidants are compounds that slow down and/or
added. The sample then is titrated with NaOH and the
inhibit oxidation processes that degrade foods and other
percent FFA is calculated.
materials. Antioxidants like butylated hydroxyanisole
(BHA) are added to food to prevent fats from becoming
x 100
rancid and to minimize decomposition of vitamins and
essential fatty acids; they work by scavenging destructive
Importance of measuring %FFA:
free radicals from foods.
 In crude fat, FFA or acid value estimates the amount
of oil that will be lost during refining steps designed
to remove fatty acids. 3. What is peroxide value? Acid value?
 In refined fats, a high acidity level means a poorly  Peroxide value is a measure of the peroxides contained
refined fat or fat breakdown after storage or use. in the oil and is usually determined by volumetric
However, if a fat seems to have a high amount of methods.
FFAs, it may be attributable to acidic additives (e.g.,
 Acid value (AV) is defined as the mg of KOH necessary to
citric acid added as a metal chelator) since any acid
neutralize the free acids present in 1 g of fat or oil. The
will participate in the reaction.
AV is often used as a quality indicator in frying oils,
o If the fatty acids liberated are volatile, FFA
where a limit of 2mg KOH/g oil is sometimes used.
or acid value may be a measure of
%FFA(as oleic) × 1.99 = acid value
hydrolytic rancidity.
4. You want to compare several fat/oil samples for the
Exercise 8: Determination of Peroxide Value
chemical characteristics listed below. For each
Fat undergo changes during storage which result in the characteristic, name one test that can be used to obtain
production of an unpleasant taste and odor, which is information desired:
commonly known as rancidity. a. Degree of unsaturation Iodine value
o Rancidity is caused by the action of air (oxidative) or b. Predicted susceptibility to oxidative rancidity
by microorganisms (ketonic). Schaal oven test
c. Present statues with regard to oxidative rancidity
Anisidine value and Totox value
d. Average fatty acid molecular weight 
Saponification number
Principle of Methods
e. Amount of solid fat at various temperatures Solid
To a known amount of fat or oil, excess potassium iodide is
added, which reacts with the peroxides in the sample. The fat index
iodine liberated is titrated with standardized sodium f. Hydrolytic rancidity Free Fatty Acid
thiosulfate using a starch indicator. The calculated amount of
potassium iodide required to react with the peroxide present 5. What methods would be useful in determining the
is used to determine the peroxide value. effectiveness of various antioxidants added to an oil?
 Oxygen Radical Absorbance Capacity (ORAC)-
SELF TEST
measures antioxidant inhibition of peroxyl radical
1. List 10 chemical compounds that are permitted to be used
induced oxidations and thus reflects classical radical
antioxidants in foods. chain breaking antioxidant activity by H atom
1). Propyl Gallate (PG) transfer
2). t-butylhydroquinone (THBQ)  Total Radical-Trapping Antioxidant Parameter
3). Ionox 100 (TRAP)- involves the initiation of lipid peroxidation
4). Butylated Hydroxytoluene (BHT) by generating water-soluble peroxyl radicals and is
5). Dodecyl Gallate (DG) sensitive to all known chain breaking antioxidants,
but it is relatively complex and time-consuming to
6). 2,4,5-trihydroxybutyrophenone (THBP)
perform, requiring a high degree of expertise and
7). Octyl Gallate (OG)
experience.
8). Nordihydroguaiaretic Acid (NDGA)
 Total Oxidant Scavenging Capacity (TOSC)- permits
9). Ascorbyl Palmitate (AP)
quantification of the absorbance capacity of
10). 2 (or 3)-tert-butyl-4-hydroxyanisole (BHA)
FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 10
 antioxidants specifically toward three potent Principle of the Method
oxidants, that is, hydroxyl radicals, peroxyl radicals, This method is based on the determination of the volumes of
and peroxynitrite. test solution required to reduce completely a known volume
 Chemiluminescence (CL)- based on the reaction of of an alkaline solution of copper. The endpoint is indicated by
radical oxidants with marker compounds to produce titration with methylene blue.
excited state species that emit chemiluminescence
(chemically induced light). Any compounds that
react with the initiating radicals inhibit the light
production.
 2,2-Diphenyl-1-picrylhydrazyl (DPPH) Assay - based
on the measurement of the reducing ability of
antioxidants toward DPPH¥. The ability can be
1. Compare the result of Fehling’s test with sugar? With invert
evaluated by electron spin resonance (EPR) or by
measuring the decrease of its absorbance. sugar? Explain your answer.
 The test is commonly used for reducing sugars but is
6. List the factors which affect the onset of rancidity. known to be NOT specific for aldehydes. Table sugar or
 Oxidative rancidity is accelerated by exposure to sucrose will have a negative result because it is a non-
heat (temperature), light, moisture and presence of reducing sugar. Whereas, invert sugar, consisting of
trace metals. Other factors include chemical fructose and glucose, will give a positive result since both
structure of fat and presence of contaminants. are reducing sugars.

7. A 5 g of food- grade oil was reacted with excess KI to

2. Why are Fehling’s A and Fehling’s B kept separate?
determine peroxide value. The free iodine was titrated with
 Fehling’s A contains and Fehling’s B are kept separate to
a standardized solution of 0.1 N sodium thiosulfate. The
prevent deterioration, i.e. conversion of Cu(OH)2 to black
amount of titrant was 0.6 ml (blank corrected). Calculate
precipitate of CuO. Sodium potassium tartarate is added
the peroxide value of the oil.
to prevent precipitation of cupric hydroxide.

3. What is the difference of reducing sugar and non- reducing

8. A 10 g sample vegetable oil was used to determine free
sugar?
fatty acid. One ml of 0.01 N NaOH was used to titrate the
 The primary difference between reducing and
sample to endpoint. What is the acid value of the sample?
nonreducing sugars is the presence of a free group in the
prior. Sugars that can be oxidised by mild oxidising
agents are called reducing sugars because the oxidising
agent is reduced in the reaction. A non-reducing sugar is
Exercise 9: Determination of Sugars not oxidised by mild oxidising agents.

Sugars may be determined by chemical, enzymatic, and HPLC 4. How does sugar lengthen shelf- life of processed fruit?
techniques.  The principle of this technology is to add sugar in a
 The chemical methods generally involve reaction of quantity that is necessary to augment the osmotic
reducing sugars with copper (III) in an alkaline pressure of the product's liquid phase at a level
which will prevent microorganism development.
solution.
 From a practical point of view, however, it is usual to
 The color change associated with the reaction is partially remove water (by boiling) from the product
believed to be due to enolization, with resultant to be preserved, with the objective of obtaining a
double- bond formation producing color. higher sugar concentration. In concentrations of 60%
in the finished products, the sugar generally assures
Factors affecting the reaction includes: food preservation.
a. Type of sugar
5. A 5 g sample of jam is made up to 250 ml and gives a titer
b. Concentration of sugar
of 26.8 ml using 10 ml of mixed Fehling’s solution. Compute
c. Concentration of alkali
for % reducing sugar.
d. Temperature
e. Time

FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 11
 Exercise 11: Determination of Chloride 3. Would Mohr titration procedure be applicable to
Sodium content of foods can be determined by various determine the salt content of grape juice or catsup? Why
methods, including an ion selective electrode (ISE), the Mohr or why not?
or Volhard titration procedure, or indicator test strips.  Mohr’s method works in the pH region of 6-9 only. To make
 All these methods are faster and less expensive it applicable to grape juice or catsup, an adjustment in their
procedures than analysis by atomic absorption pH must be performed prior. A pH above 6-9, will result to
spectroscopy or inductively coupled plasma-atomic a precipitation between silver and hydroxide. Below this pH
emission spectroscopy. chromate converts to dichromate, a bright orange colour
thereby obscuring the endpoint. Calcium carbonate is
Principle of Method often added to reduce the acidity of the solution.
The Mohr titration is a direct titration method to quantitate
chloride ions, to then calculate sodium ions. The chloride- 4. Would overshooting the endpoint result in an over- or
containing sample solution is titrated with a standard solution underestimation of the salt content using Mohr titration?
of silver nitrate. After the silver from silver nitrate has
complexed with all the available chloride in the sample, the  Overshooting the endpoint will result to an overestimation
silver reacts with chromate that has been added to the of the salt content since the increase in volume of the silver
sample, to form an orange- colored solid, silver chromate. nitrate would directly increase the computation of % NaCl.
The volume of silver used to react with the chloride is used to
calculate the sodium content of the sample.

SELF TEST

1. Show the calculation of how to prepare ½ liter of 0.1 M

AgNO3.
MW AgNO3= 169.87 g/mol
M= 0.1 M
L= 0.5 L

Wt. sample= M x L solution x MW AgNO3

= 0.1 M x 0.5 L x 169.87 g/mol
Wt. sample= 8.4935 g

 Weigh 8.4935 g silver nitrate and dissolved to 500 ml

to produce ½ liter of 0.1 M AgNO3.

2. Show the reaction taking place to determine NaCl by

titration.
As the silver nitrate solution is slowly added, a precipitate of
silver chloride forms.
+ -
Ag (aq) + Cl (aq) --> AgCl(s)

The indicator used is dilute potassium chromate solution.

When all the chloride ions have reacted, any excess silver
nitrate added will react with chromate ions to form a red-
brown precipitate of silver chromate. This procedure is
known as Mohr’s method.
2Ag+(aq) + CrO42-(aq) --> Ag2CrO4(s)

FT 105 Lab /Ang tulog madaling bawiin, ang isang sem hindi/ Page 12