Practicum 5

Moisture analysis
A distillation method

Outline
Learning objectives.................................................................................................................... 1
Introduction ............................................................................................................................... 1
Quiz............................................................................................................................................ 4
Case study.................................................................................................................................. 4

Learning objectives
1. Understand the principle of moisture analysis using distillation method
2. Enable to calculate, analyze and interpret data of moisture analysis

Introduction
Distillation techniques involve codistilling the moisture in a food sample with a high boiling
point solvent that is immiscible in water, collecting the mixture that distills off, and then
measuring the volume of water. Two distillation procedures are in use today: direct and reflux
distillations, with a variety of solvents. For example, in direct distillation with immiscible
solvents of higher boiling point than water, the sample is heated in mineral oil or liquid with
a flash point well above the boiling point for water. Other immiscible liquids with boiling point
only slightly above water can be used (e.g., toluene, xylene, and benzene). However, reflux
distillation with the immiscible solvent toluene is the most widely used method.

Distillation techniques were originally developed as rapid methods for quality control work,
but they are not adaptable to routine testing. The distillation method is an AOAC-approved
technique for moisture analysis of spices (AOAC Method 986.21), cheese (AOAC Method
969.19), and animal feeds (AOAC Method 925.04). It also can give good accuracy and precision
for nuts, oils, soaps, and waxes.
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Distillation methods cause less thermal decomposition of some foods than oven drying at high
temperatures. Adverse chemical reactions are not eliminated but can be minimized by using
a solvent with a lower boiling point. This, however, will increase distillation times. Water is
measured directly in the distillation procedure (rather than by weight loss), but reading the
volume of water in a receiving tube may be less accurate than using a weight measurement.

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Reflux distillation uses either a solvent less dense than water (e.g., toluene, with a boiling
point of 110.6◦C; or xylene, with a boiling range of 137–140◦C) or a solvent denser than water
(e.g., tetrachloroethylene, with a boiling point of 121◦C). The advantage of using this last
solvent is that material to be dried floats; therefore, it will not char or burn. In addition, there
is no fire hazard with this solvent.

A Bidwell–Sterling moisture trap (Fig. 1) is commonly used as part of the apparatus for reflux
distillation with a solvent less dense than water. The distillation procedure using such a trap
is described below, with emphasis placed on dislodging adhering water drops, thereby
minimizing error. When the toluene in the distillation just starts to boil, the analyst will
observe a hazy cloud rising in the distillation flask. This is a vaporous emulsion of water in
toluene. Condensation occurs as the vapors rise, heating the vessel, the Bidwell–Sterling trap,
and the bottom of the condenser. It is also hazy at the cold surface of the condenser, where
water droplets are visible. The emulsion inverts and becomes toluene dispersed in water. This
turbidity clears very slowly on cooling.

Three potential sources of error with distillation should be eliminated if observed:

1. Formation of emulsions that will not break. Usually this can be controlled by allowing
the apparatus to cool after distillation is completed and before reading the amount of
moisture in the trap.
2. Clinging of water droplets to dirty apparatus. Clean glassware is essential, but water
seems to cling even with the best cleaning effort. A burette brush, with the handle
end flattened so it will pass down the condenser, is needed to dislodge moisture
droplets.
3. Decomposition of the sample with production of water. This is principally due to
carbohydrate decomposition to generate water (C6H12O6 → 6H2O + 6C). If this is a
measurable problem, discontinue method use and find an alternative procedure.

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Supplies and Equipment

1. Sample: basil leave, shallot (fresh)
2. Toluene

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 3. Analytical balance, 0.1 mg sensitivity
4. Glass distillation apparatus with ground glass joints: (1) Boiling flask, 250 ml or 300
ml, roundbottom, shortneck flask with a T.S. 24/40 joint; (2) West condenser with drip
tip, 400 mm in length with a T.S. 24/40 joint; (3) Bidwell–Sterling trap, T.S. 24/40 joint,
3-ml capacity graduated in 0.1 ml intervals.
5. Heat source, capable of refluxing toluene in the apparatus above (e.g., heating mantle
connected to voltage controller). No open flame!
6. Nylon bristle buret brush, ½ in. in diameter, and a wire loop. (It should be long enough
to extend through the condenser, ca. 450 mm. Flatten the loop on the buret brush
and use this brush, inverted, as a wire to dislodge moisture drops in the moisture
trap.)

Hazards, Cautions, and Waste Disposal

CAS No. Hazards
Toluene 108-88-3 Harmful, highly flammable
Toluene is highly flammable and is harmful if inhaled. Use adequate ventilation. Wear safety
glasses and gloves at all times. For disposal of toluene waste, follow good laboratory practices
outlined by environmental health and safety protocols at your institution.

Work Procedures
General method
1. Place sample in distillation flask and cover completely with solvent.
2. Fill the receiving tube (e.g., Bidwell-Sterling Trap) with solvent, by pouring it through
the top of the condenser.
3. Bring to a boil and distill slowly at first then at increased rate.
4. After the distillation has proceeded for approximately 1 hr, use an adapted buret
brush to dislodge moisture droplets from the condenser and top part of the Bidwell-
Sterling trap.
5. Slide the brush up the condenser to a point above the vapor condensing area.
6. Rinse the brush and wire with a small amount of toluene to dislodge adhering water
drops.
7. If water has adhered to the walls of the calibrated tube, invert the brush and use the
straight wire to dislodge this water so it collects in the bottom of the tube.
8. Return the wire to a point above the condensation point, and rinse with another small
amount of toluene.
9. After no more water has distilled from the sample, repeat the brush and wire routine
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to dislodge adhering water droplets.

10. Rinse the brush and wire with toluene before removing from the condenser.
11. Allow the apparatus to cool to ambient temperatures before measuring the volume
of water in the trap.
12. Volume of water x 2 (for a 50 g sample) = % moisture

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AOAC official method 986.21
1. Grind the fresh basil with a small table-top food grinder. Pulse grind the sample in 5–
10 s intervals. Avoid long pulses and excessive grinding to prevent frictional heat.
2. Weigh approximately 40 g of sample (basil or NFDM) accurately (amount chosen to
yield 2–5 ml water).
3. Transfer sample quantitatively to distilling flask. Add sufficient toluene to cover the
sample completely (not less than 75 ml).
4. Assemble the apparatus as shown in Fig. 1. Fill the trap with toluene by pouring it
through the condenser until it just fills the trap and begins to flow into the flask. Insert
a loose non absorbing cotton plug into the top of the condenser to prevent
condensation of atmospheric moisture in the condenser.
5. Bring to boil and reflux at about two drops per second until most of the water has
been collected in the trap, then increase the reflux rate to ca. four drops per second.
6. Continue refluxing until two consecutive readings 15 min apart show no change.
Dislodge any water held up in the condenser with a brush or wire loop. Rinse the
condenser carefully with ca. 5 ml toluene. Dislodge any moisture droplets adhering to
the Bidwell– Sterling trap or toluene trapped under the collected moisture. For this,
use the wire. Rinse wire with a small amount (10 ml) of toluene before removing from
apparatus.
7. Continue refluxing for 3–5 min, remove the heat, and cool the trap to 20°C in a
suitable water bath.
8. Calculate the moisture content of the sample:
% Moisture = [vol. of water ml /wt of sample g] x 100

Data Quality
The data quality might be evaluated using RSD and CV Horwits value. Repeatability is good if
RSD<2/3CV Horwitz. RSD is Relative Standard Deviation which might be calculated as follow:

SD x 100
𝑅𝑆𝐷 =
𝑥̅

Where, SD is Standard Deviation, and 𝑥̅ is average value.

While CV Horwits can be calculate using this formula:

𝐶𝑉 𝐻𝑜𝑟𝑤𝑖𝑡𝑠 = 2(1 − 0.5 log 𝐶)

Quiz
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The quiz can be accessed via the following link:

Case study
The simulation video can be accessed via the following link:
https://drive.google.com/drive/u/3/folders/1TNvULH1CL5qtBndwEdZ4Eog1DlO6udRZ

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Moisture Analyses
Sample (Onion) Sample 2 (Basil Leave)
Repetition X (g) Y (mL) Repetition X (g) Y (mL)
1 10.1121 8.1 1 10.0578 6.5
2 10.1098 7.3 2 10.0766 6.5
3 10.0213 6.8 3 10.0167 6.4

1. In each case below, would you likely overestimate or underestimate the moisture
content of a food product being tested? Explain your answer! (a) Emulsion between
water in sample and solvent not broken. (b) Water clinging to condenser.
2. Identify the key points in the procedure of moisture analysis of onion and basil that
should need to be considered to assure the data quality!
3. How to check the data quality?
4. According to the above result please check the data quality and interpret the result!

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