This document reports on experiments conducted to quantify reducing and total sugars in food samples. In Experiment 1, reducing sugars were quantified using redox titration with ferricyanure solution. The volume of sample and glucose standard required to react with the ferricyanure allows calculating the reducing sugar content. In Experiment 2, total sugars were quantified by first hydrolyzing samples with hydrochloric acid to break down polysaccharides, then performing redox titration and calculation. Coconut water and banana were analyzed, with banana requiring ethanol extraction first. Reducing and total sugar contents were calculated and reported.
This document reports on experiments conducted to quantify reducing and total sugars in food samples. In Experiment 1, reducing sugars were quantified using redox titration with ferricyanure solution. The volume of sample and glucose standard required to react with the ferricyanure allows calculating the reducing sugar content. In Experiment 2, total sugars were quantified by first hydrolyzing samples with hydrochloric acid to break down polysaccharides, then performing redox titration and calculation. Coconut water and banana were analyzed, with banana requiring ethanol extraction first. Reducing and total sugar contents were calculated and reported.
This document reports on experiments conducted to quantify reducing and total sugars in food samples. In Experiment 1, reducing sugars were quantified using redox titration with ferricyanure solution. The volume of sample and glucose standard required to react with the ferricyanure allows calculating the reducing sugar content. In Experiment 2, total sugars were quantified by first hydrolyzing samples with hydrochloric acid to break down polysaccharides, then performing redox titration and calculation. Coconut water and banana were analyzed, with banana requiring ethanol extraction first. Reducing and total sugar contents were calculated and reported.
This document reports on experiments conducted to quantify reducing and total sugars in food samples. In Experiment 1, reducing sugars were quantified using redox titration with ferricyanure solution. The volume of sample and glucose standard required to react with the ferricyanure allows calculating the reducing sugar content. In Experiment 2, total sugars were quantified by first hydrolyzing samples with hydrochloric acid to break down polysaccharides, then performing redox titration and calculation. Coconut water and banana were analyzed, with banana requiring ethanol extraction first. Reducing and total sugar contents were calculated and reported.
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VIETNAM NATIONAL UNIVERSITY – HO CHI MINH CITY
HO CHI MINH CITY UNIVERSITY OF TECHNOLOGY
EXPERIMENT REPORT SUBJECT: LABORATORY OF FOOD CHEMISTRY AND BIOCHEMISTRY LAB 8: CARBOHYDRATE - QUANTIFICATION
Class: CC01 Group: 04 Instructor: Assoc. Prof. Dr. VO DINH LE TAM Group members: Huỳnh Diệp Mỹ Phụng 2011869 Đặng Thụy Huỳnh Như 2011771 Nguyễn Cao Ngọc Quý 2053389
1 EXPERIMENT 8.1: QUANTIFICATION OF REDUCING AND TOTAL SUGAR 1. Quantification of reducing sugar by the method for redox titration with ferricyanure 1.1. Objective Using the method of redox titration with ferricyanure to determine the amount of reducing sugar in a solution. 1.2. Principle Ferricyanure K 3 Fe(CN )6 reacts with reducing sugar, forming ferrocyanure. Using this reaction can determine the amount of reducing sugar in solution. Titration is done in alkaline condition (NaOH) when heated with a methylene blue indicator. The reaction equation is as follows:
This method is simpler than the method of using alkaline solution of copper sulfate due to not producing precipitate and the signal of the reaction ending easily being seen. Result calculation does not base on theoretical equations, instead, using experimental formulas. The precision of results depends on many factors, but experimental procedure and performance are the most important. All monosaccharides and some oligosaccharides are reducing sugars. Oligosaccharide and polysaccharides are hydrolyzed easily to form monosaccharides, so we can quantity reducing sugar before and after hydrolysis to determine their content. 1.3. Apparatus - Electric stove, clamp, wire gauze, water bath. - Filter funnel, graduated cylinder, 100ml volumetric flask, beaker, conical flask, burette, pipette. 1.4. Materials and chemicals - Coconut water: 2 groups - Banana: 2 groups - 1% K 3 Fe(CN )6 solution - 0.5% glucose (w/v) - 5% and 2.5N NaOH solution - 5% HCl
2 - 10% CCl3 COOH - 1% Methyl red - 0.04% Methylene blue 1.5 Procedure - Sample contains much starch or inulin (banana) + Extract sugar by Ethanol 70-80% + Grind sample, add the ethanol, use a pot containing water to heat the mixture being contained in a round flask (assembled with an air condenser). In this case, it is unnecessary to precipitate protein because the amount of protein in the mixture is not remarkable). Titration of reducing sugar - Pour sample containing reducing sugar (after filtration) into a burette (note the origial volume of the sample in the burette to calculate the used Vk). - Put 10ml of 1% K 3 Fe(CN )6 solution and 2.5ml of 2.5N NaOH solution into an Erlenmeyer flask/conical flask. - Boiling and titrate on the electric stove using reducing sugar solution from the burette (drop one by one, shake strongly) - Initial solution has lime yellow of ferricyanure. The time to stop the titration is when the lime yellow disappears, the solution is transparently colorless in 30 seconds and becomes the light straw-yellow of ferrocyanure. - In the case that it is difficult to realize the time when color changes, to determine the titration stop point, drop a droplet of methylene blue and a first extra drop of sugar will make the blue disappear and the reaction finished. - The result of the first titration is just for reference for the second titration. At the second titration, after boiling ferricyanure solution, quickly drop the reducing sugar solution (based on the first result), leaving about 1ml of the solution to titrate continuously to determine the titration stop point. - When calculating, use data from the second titration time afterwards. - Perform a similar experiment for 0.5% glucose standard solution to get the volume of 0.5% glucose solution to titrate. 1.6 Calculation In the experiment, both Vk (ml) of sample solution and Vg (ml) of 0.5% glucose react with the amount of ferricyanure solution (at a determined/known concentration).
3 So, in Vk ml of sample solution or in Vg ml of 0.5% glucose solution, there is (0.5 x Vg) / 100g of glucose. The amount of reducing sugar is calculated using the following formula: 0.5 ×Vg × V × 100 Xk = 100× Vk × m
Where Xk: The amount of reducing sugar, g/100g or g/100mL Vg: The volume of 0.5% glucose solution to titrate, ml Vk: The volume of reducing sugar to titrate, ml V: The volume of volumetric flask, ml m: The weight of sample used, g or ml 1.7 Result and report
Parameter Result
The volume of 0.5% glucose solution to titrate, ml, (Vg) 27ml
The volume of reducing sugar to titrate, ml, (Vk) 2ml
The volume of volumetric flask, ml, (V) 100ml
The weight of sample used, g or ml, (m) 6.32g
The amount of reducing sugar, g/100g or g/100mL, (Xk) 0.5 ×Vg × V × 100 Xk = 100× Vk × m 0.5× 27 ×100 ×100 ¿ 100 ×2 ×6.32 = 106.804 g/100mL
2. Quantification of total sugar
2.1. Objection Quantify extractable soluble carbohydrates in water of a food sample. 2.2. Principle Total sugar includes extractable soluble carbohydrates in water. Complex carbohydrate 4 contains a large number of sugar molecules which are bonded together with glycosidic linkage. Using hydrochloric acid to hydrolyze oligosaccharides and polysaccharides into monosaccharides, then quantify total sugar content in a food sample using ferriccyanure. 2.3. Apparatus - Electric stove, clamp, wire gauze, water bath - Filter funnel, graduated cylinder, 100ml volumetric flask, beaker, conical flask, burette, pipette 2.4. Materials and chemicals - Banana: 2 groups - 1% K 3 Fe(CN )6 solution - 0.5% Glucose (w/v) - 5% and 2.5N NaOH solution - 5% HCl - 10% CCl3 COOH - 1% Methyl red - 0.04% Methylene blue 2.5. Procedure - Pour wxactly 50ml of the above reducing sugar solution into a 100mL vlumetric flask. Add 20ml of 5% HCl solution, heat the mixture in 30-45 minutes with a water containing pot. - Cool fast, neutralize the mixture by 2.5N NaOH solution or saturated Na2 SO3 solution using litmus paper. - Add water to the line. - Do the titration the same as above: titration 0.5% standard glucose (to get Vg) and titration with the total sugar sample (to get Vt) 2.6. Calculation The content of total sugar is calculated by the following formula: 0.5 ×Vg ×V 1 × V 2 × 100 Xt = 100 ×Vt ×50 × m Where Xt : the content of total sugar, % Vg : volume of 0.5% glucose solution for titrating, ml Vt : volume of total sugar solution for titrating, ml V 1: volume of volumetric flask of solution needed to determine reducing sugar content, ml V 2: volume of volumetric flask of solution needed to determine total sugar, ml m: weight of sample used, g or ml
5 2.7. Result and report Parameter Result The volume of 0.5% glucose solution for 27 titrating, ml The volume of total sugar solution for 7.5 titrating, ml The volume of volumetric flask of solution 100 needed to determine reducing sugar content, ml The volume of volumetric flask of solution 100 needed to determine total sugar, ml The weight of sample used, g or ml 6.83 The content of total sugar, % 0.5 ×Vg ×V 1 × V 2 × 100 Xt = 100 ×Vt ×50 × m 0.5 ×27 ×100 ×100 ×100 Xt = 100 ×7.5 ×50 ×6.83 ≈ 52.7086 %
3. Questions
Present principle of method for quantifying reducing sugar using ferricyanure
Reducing sugars undergo oxidation in presence of an oxidizing agent. This property has been exploited for their analysis since quite some time. Present method involves estimation of reducing sugars using Cole's ferricyanide method.
Under boiling circumstances, Cole's ferricyanide technique includes titrating an alkaline
solution of ferricyanide ions Fe(CN)63- with reducing sugars. The change in oxidation state of iron from Fe(III) to Fe(IV) marks the transition from ferricyanide to ferrocyanide (II). The color change from yellow to colorless, which is difficult to identify with the human eye, indicates complete conversion from ferricyanide to ferrocyanide. A redox electrode is used to detect the change in potential that occurs as a result of this transition.
Predict some errors that may occur in the experiment
Error in neutralizing the mixture in quantification of total sugar. If we don’t
carefully drop NaOH solution, the mixture will become over base.
6 The final results can be affected by the environment factor such as the heat that we boil the solution or the boiling time. The weight of the sample can be decreased during the process that affects the final results and calculation. Difficulty in determining the time when color changes, we can observe the wrong number.
Present some properties of monosaccharide
A monosaccharide or simple sugar is a carbohydrate that cannot be hydrolyzed into
smaller carbohydrates. Like all carbohydrates, a monosaccharide consists of three chemical elements: carbon, hydrogen, and oxygen. Monosaccharides include aldoses, ketoses, and their derivatives. The general chemical formula for a monosaccharide is
CnH2nOn or (CH2O)n.
In pure form, monosaccharides are crystalline, water-soluble, colorless solids.
Monosaccharides have a sweet flavor because the orientation of the OH group interacts with the taste receptor on the tongue that detects sweetness. Via a dehydration reaction, two monosaccharides can form a disaccharide, three to ten can form an oligosaccharide, and more than ten can form a polysaccharide.
Monosaccharides serve two main functions within a cell. They are used to store and produce energy. Glucose is a particularly important energy molecule. Energy is released when its chemical bonds are broken. Monosaccharides are also used as building blocks to form more complex sugars, which are important structural elements.
4. References David, L. N., & Michael, M. (2017). Cox, Principles of biochemistry (7th Edition). W. H. Freeman and Company, 3270 pages. Najafpour, G. (2015). Biochemical engineering and biotechnology (second edition). Elsevier, 650 pages. Srinivasan, D., & Kirk, L. P. (2017). Fennema's Food Chemistry (5th Edition). CRC press, 1107 pages. Trần, B. L., Tôn, N, M, N., & Định, T. N. T. (2011). Thí nghiệm Hóa sinh Thực phẩm, ĐHQG TP.HCM, 83 trang.
