manual of methods for determining micronutrients in fortified foods

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4. Add 10 mL of n-heptane and a small volume of saturated saline solution to improve separation. Shake in a vortex mixer for 1 minute using a stopwatch. 5. Centrifuge the tube for 10 minutes at 3000 rpm. A clear separation between the solids-aqueous layer and the organic layer must be obtained. The organic layer will be yellow (see picture below). 6. Using a Pasteur pipette, transfer the organic phase (~10mL) to a clean test tube (See test tubes with the yellow solution in the pictures shown below in step 8). 7. Using a pipette, transfer 2 mL of the organic extract to another test tube. 8. Pipette 3 mL of the TFA solution and add them fast and vigorously into the tube containing the extract, so the addition will mix the solution. The development of a blue color indicates the presence of vitamin A in the flour and the intensity of the color is directly proportional to the concentration of vitamin A in the sample. The pictures below show different shades of blue in two samples. The concentration of vitamin A in the picture to the left is higher than the concentration in the sample from picture to the right. 9. If a blue or light blue color is observed, report the results as positive and above 0.5 mg/kg. - 24 -
VIII. Qualitycontrol Run the flour control with a known concentration of vitamin A every time wheat flour samples are analyzed. If the reaction is negative with the control, check the procedure and the stability of the TFA solution. - 25 -
Page 1 and 2: MANUAL OF METHODS FOR DETERMINING M
Page 3 and 4: - 3 -
Page 5 and 6: I. Introduction Food fortification
Page 7 and 8: A. Qualitative method to determine
Page 9 and 10: VII. Interpretation • Unfortified
Page 11 and 12: − − − − − − Refrigerato
Page 13 and 14: 3. Stir with a magnetic stirrer for
Page 15 and 16: IX. Calculations 1. Plot a graph of
Page 17 and 18: − 1,10-phenanthroline-monohydrate
Page 19 and 20: . Preparation of the ash solution 1
Page 21 and 22: III. Methods to determine vitamin A
Page 23: IV. Equipment and materials − 50
Page 27 and 28: − Volumetric flasks (100 and 10 m
Page 29 and 30: VII. Procedure a. Preparation of sa
Page 31 and 32: c. Clean-up and concentration proce
Page 33 and 34: VIII. Calculations Results can be c
Page 35 and 36: IV. Methods to determine water-solu
Page 37 and 38: A. Method to determine riboflavin i
Page 39 and 40: VI. Standard solutions 1. Riboflavi
Page 41 and 42: 8. Pipette exactly 4.0 mL of the fi
Page 43 and 44: B. Determination of thiamin in flou
Page 45 and 46: In a 25-mL beaker weigh 0.9602 g so
Page 47 and 48: C. Determination of niacin in flour
Page 49 and 50: 7. Transfer the extract to a 250 mL
Page 51 and 52: IV. Equipment − Analytical balanc
Page 53 and 54: 6. Ammonium hydroxide-0.1M: In a 10
Page 55 and 56: VIII. Procedure A. Strain propagati
Page 57 and 58: Note: Prior to start analyzing samp
Page 59 and 60: IX. Calculations 1. Calculate the a
Page 61 and 62: B. Verification 1. In an Erlenmeyer
Page 63 and 64: V. References • Bowman BA and Rus

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