20th Dec. 2009

B.K.K.K. Jinadasa



Determination of fibre
4.0 Determination of fibre 4.1 Determination of crude fibre 4.1.1. Introduction Crude fibre is the insoluble and combustible organic residue which remains after the sample has been treated under prescribed conditions. The most common conditions are consecutive treatments with light petroleum, boiling dilute sulphiuric acid, boiling dilute sodium hydroxide, dilute hydrochloric acid, alcohol and ether. This treatment provides a crude fibre consisting largely of the cellulose content together with a proportion of the lignin and hemicelluloses content of the sample. The amounts of these substances in the crude fibre will vary with the conditions employed. 4.1.2. Materials and reagents Round bottom flask Beaker Reflux condenser Sintered crucible (Gooch crucible) Asbestos wool Desiccator Heating mantle Weighing balance Sulphuric acid dil (H2SO4) Sodium hydroxide (NaOH) Litmus paper Sample tea

4.1.3. Procedure 2g of defatted sample (tea) was weighed and transferred into a 1 L conical flask. 200 mL of boiling 1.25% H2SO4 was added into the flask and boiled for 30 minutes over a Bunsen burner. Occasional swirling was done to remove solids from the sides of the flask. After 30 minutes solution was decanted through a Buchner funnel (filtered by Whatman 52 filter paper). All the residues were transferred and rinsed with boiling water. Then the residue was transferred from filter paper into the flask by using 200 mL of 1.25% W/V NaOH solution which was near to boil. Contents in the flask were boiled as quickly as possible for 30 minutes. Boiling water was added to maintain the volume. After this, solution was filtered through a rapid hardened filter paper. Residue was washed with 1% HCl until the filtrate was free from alkali. Then the residue was washed with small amounts of alcohol (15 mL) and diethyl ether (10 mL) respectively. Residue was transferred quantitatively to a tared crucible and dried in the oven at 105°C for 2 hours. After 2 hours sample was cooled and weight was taken. Then the sample was incinerated at 500°C, cooled and weight was taken.


Determination of fibre
4.1.4. Results and calculation Weight of tea sample (m1) Moisture % tea sample Dry weight of tea sample (m1) Weight of crucible (m2) Weight of crucible + filter paper (m3) Weight of crucible + filter paper + fibre residue (m4) Weight of crucible + ash (m5) = 2.0136 g = 7.0773% = 1.8711 g = 30.2048 g = 30.7822 g = 30.9946 g = 30.2060 g

= = =

(m4-m3) (m5-m2) × 100 m1 (30.9946-30.7822) g (30.2060-30.2048) g × 100 1.8711 g 11.29 %

4.1.5. Discussion Apparent crude fibre is lowered by defatting, though the low lipid content of some foods affects the results little. Sample should be defatted using ether, before doing acid and alkaline digestion. In this procedure carbohydrates is removed using H2SO4 digestion and protein is removed by using NaOH digestion. In acid digestion, boiling is to be taken within one minute; therefore H2SO4 was boiled and then added to the flask. Before digestion with NaOH all the residue in the Buchner funnel should be transferred carefully to the digestion flask. For this previously boiled NaOH is used. To get a constant result, the boiling technique must be visibly, carefully followed. After digesting in NaOH, the residue should be filtered with pre weighed filter paper; crucible and filter paper were measured before the NaOH digestion. After filtering, the residue was washed with 01 % HCl to get rid of all the NaOH remain in the residue. Then it should be washed with distilled water to remove all HCl. Then washed with C2H5OH to remove chlorophylls and gums. Filtering after each digestion must be completed within a given time, delays in filtering generally lower the results. The finer the sample material is ground, lower the fibre content. According to the SLSI standards (SLS 135:2009), max crude fibre % by mass (max) should be 16.5%.


Determination of fibre
4.2 Determination of acid detergent fibre 4.2.1. Introduction An acidified detergent solution is used to dissolve cell solubles, hemicellulose and soluble minerals leaving a residue of cellulose, lignin, and heat damaged protein and a portion of cell wall protein and minerals (ash). 4.2.2. Materials and reagents Volumetric flasks (100mL, 200mL) Measuring cylinder Heating mantle Sintered glass crucible Refluxing system Desiccator Sulphuric acid Cetyl trimethyl ammonium bromide solution (5 g of cetyl trimethyl ammonium bromide dissolved in 250 mL of 2M sulphuric acid) Sample tea

4.2.3. Procedure 1g of air dried tea sample was weighed and transferred to a refluxing flask. 100 ml of acid detergent solution was added into it. Then the flask was heated to boil and refluxed for 1 hour. After 1 hour sample was allowed to filter through a pre weighed glass crucible using minimum suction. Sample was washed twice with hot water and then washed with acetone until no more colour is removed. Then the sample was dried for three hours at 100ºC. 4.2.4. Results and calculation Weight of sample Moisture percentage Weight of dry sample Weight of glass crucible Weight of crucible + fibre after drying = 1.0014 g = 7.0773 % = 0.9305 g = 136.8328 g = 137.1034 g

= 137.1034 136.8328 × 100 0.9305


Determination of fibre
= 29.08 % 4.2.5. Discussion The content of fibre extracted using the acid detergent method was significantly higher than the fibre content extracted by crude fibre method. During the filtering through glass crucible, low suction was applied. If the suction is very high, the fibre particles may be penetrating in to the pores of the filter


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