The purpose of the document is to establish a general and uniform method for the determination of total dietary fiber in foods.
2.0 SCOPE
This standard testing procedure is suitable for the determination of total dietary fiber in food samples.
3.0 RESPONSIBILITY
The expert and trained chemist are responsible for following this procedure. The technical manager is responsible for implementing this procedure.
4.0 REQUIREMENTS
4.1 Apparatus
4.1.1 Water bath
4.1.2 pH Meter
4.1.3 Analytical balance
4.1.4 Desiccator
4.1.5 Muffle furnace
4.1.6 Hot air oven
4.2 Glassware
4.2.1 G-2 crucible
4.2.2 Beaker
4.2.3 Measuring Cylinder
4.3 Reagent
4.3.1 NaOH ( 0.275 M)
4.3.2 HCl( 0.325 M)
4.3.3 Phosphate buffer(0.08M)
4.3.4 Protease enzyme
4.3.5 Amyloglucosidase
4.3.6 Alpha-amylase
4.3.7 Ethanol (95% and 78%)
4.3.8 Acetone
5.0 PROCEDURE
5.1. Weigh 1g sample in four 100 ml beakers.
5.2. Take 2 empty beaker and run blank in the same manner as in test sample (leaving sample weight follow the whole procedure in blank as well).
5.3. Add 50 ml phosphate buffer in each.
5.4. Add 0.1 ml amylase in each beakers &cover each beaker with aluminium foil..
5.5. Keep the beakers on the boiling water bath and keep on shaking, after 5 minute interval till the internal temp. of the solution reaches 95°C-100°C.
5.6. Allow to cool to room temp.
5.7. Adjust the pH to 7.5 + 0.2by adding 10 ml of 0.275 N NaOH to each beaker.
5.8. Check pH and adjust if necessary, using NaOH or HCl.
5.9. Add 0.1 ml protease enzyme into each beaker (50 mg/ml) in phosphate buffer).
5.10. Cover each beaker with aluminum foil & place in 60°C water bath
5.11. Continue shaking at 5-minute interval and incubate for 30 min. when the internal temp. reaches 60°C.
5.12. Allow the solution to cool to room temp.
5.13. Adjust pH to 4.0 and 4.6 by adding 10 ml of 0.325 M HCl to each beaker.
5.14. Adjust pH if necessary with NaOH/HCl.
5.15. Add 0.1 ml of amyloglucosidase to each beaker.
5.16. Cover each with aluminum foil and place in the water bath at 60°C with continuous agitation, incubate for 30 min. once the internal temp. reaches 60°C.
5.17. Add 4 volumes(280 ml) of 95% ethanol to each beaker.
5.18. Keep overnight for precipitation.
5.19. Take G-2 crucibles, add 0.5 g celite and keep at 130°C for 1 hr.
5.20. Remove the crucible &place it in the desiccator to cool and weigh it.
5.21. Wet and redistribute the bed of celite in crucible with 78% ethyl alcohol through suction.
5.22. Filter the solution through crucible,using filtration assembly.
5.23. Wash residue with three (20 ml) portion of 78% ethanol and two (10 ml) portion of 95% ethanol.
5.24. Now wash the residue with two (10 ml) portions of acetone.
5.25. The dry crucible containing residue overnight at 105°C in a hot air oven.
5.26. Cool in the desiccator and weigh all the crucibles.
5.27. Take three crucible’s containing residue for protein analysis.
5.28. Another three crucibles are transferred in the muffle for ashing at 525°C for 5 hrs.
5.29. Cool in the desiccator and weigh.
6.0 CALCULATION
Determination of blank:
B= blank, mg=weigh residue-PB - AB
Where,
Weight residue = average of residue weight (mg) for duplicate blank determination: and PB - weight (mg)of protein and ash, respectively, determined in first and second blank residues.
Calculate TDF as follows:
TDF % = [(weigh residue –P-A-B)weight test portion] X 100
Where,
Weight residue= average of weights (mg) for duplicate blank determinations: and P and A weight (mg) of protein and ash, respectively, in first and second test portion residues: and weight test portion = average of 2 test portion weight(mg) taken.
7.0 REVISION/CHANGE HISTORY: NA
0 ATTACHMENTS
· Appendix-1: FLOW CHART OF TOTAL DIETARY FIBER IN FOODS
9.0 REFERENCES
· AOAC official method 985.29
END OF THE DOCUMENT
You may like to read these links:
1. List of All SOPs and Documents for the Chemistry Laboratory
2. List of All SOPs and Documents for Quality Control
3. List of All SOPs and Documents For Quality Assurance Dept.
4. List of SOPs and Documents Required to Achieve NABL Accreditation
5. List of All SOPs and Documents for Laboratory Instruments and Equipment
Hi. I’m Writer of Researchsop.com. ’ ’ Please share these SOPs to all concern pharma people for their development. I like to fullfill the need of curious people. These things inspire me to make things looks better.
0 comments:
Post a Comment