Vitamin

 Fat-soluble vitamins
 A: Retinol or b-carotene

Vitamin analysis 


D: Cholecalciferol or Ergocalciferol
K: Phylloquinone
 E: Tocopherol
Prof. Dr. Pham Van Hung  Water-soluble vitamins
Department of Food Technology  B complex: Thiamin (B1), Riboflavin (B2), Pyridoxine (B6),
Folic acid (B9), Cyanocobalamin (B12)
 C: ascorbic acid

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Extraction Methods Extraction Methods

 Vitamin B1 and B2:
 Vitamins A, E, or D:  Boiling or autoclaving in acid plus enzyme treatment.
 Organic solvent extraction, saponification, and re-  Folate:
extraction with organic solvents. For unstable  Enzyme extraction with α-amylase, protease and γ-glutamyl
vitamins, antioxidants are routinely added to inhibit hydrolase(conjugase).
oxidation.  Niacin:
 Autoclaving in acid (noncereal products) or alkali (cereal
products).
 Ascorbic acid:
 Cold extraction with metaphosphoric acid/acetic acid..

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 Vitamin analysis 𝛽-carotene analysis
 Bioassay Methods:
1. RP-HPLC analysis was performed with the Agilent
 involving humans and animals
1100 series HPLC system (Agilent; USA).
 vitamins B12 and D.

 Microbiological Assays: 2. The Agilent TC-C18 column (5 𝜇m, 4.6 × 250 mm).
 making use of protozan organisms, bacteria, and yeast. 3. An acetonitrile-methyl alcoholethyl acetate (88 : 10 : 2)
 water-soluble vitamins. solvent. An isocratic elution and the solvent flow rate
 The growth of a certain microorganism in an extract of a was maintained at 1 mL/min.
vitamin-containing sample is compared against the growth of
this microorganism in the presence of known quantities of that 4. A diode array detector.
vitamin. 5. An UV absorbance at 453 nm.
 Physicochemical assays:
6. Injection volume: 20 μl
 include spectrophotometer, fluorometric, chromatographic,
enzymatic, immunological, and radiometric methods.

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Analysis of vitamins A, D, E, K Analysis of vitamins B

1. RP-HPLC analysis was performed with the Agilent 1. RP-HPLC analysis was performed with the Agilent
1100 series HPLC system (Agilent; USA). 1100 series HPLC system (Agilent; USA).
2. The Agilent Eclipse XDB-C18 column (5 𝜇m, 4. 6 × 2. Agilent ZORBAX Eclipse Plus C18; 250 × 4.6 mm
150 mm). i.d., 5 𝜇m).
3. Methanol solvent. An isocratic elution and the solvent 3. The isocratic delivery mobile phase (A/B 33/67; A:
flow rate was maintained at 1 mL/min. MeOH, B: 0.023 M H3PO4, pH = 3.54) at a flow rate
4. A diode array detector. of 0.5 mL/min.
5. An UV absorbance at 325 nm for vitamin A, 265 nm 4. A diode array detector.
for vitamin D3, 290 nm for vitamin E, and 244 nm for 5. An UV absorbance at 270 nm
vitamin K3. 6. Injection volume: 20 μl
6. Injection volume: 20 μl

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 Analysis of vitamins C
1. RP-HPLC analysis was performed with the Agilent
1100 series HPLC system (Agilent; USA).
2. The Agilent Eclipse XDB-C18 column (5 𝜇m, 4. 6 ×
150 mm). The end!
3. A/B 33/67; A: 0.1 M potassium acetate, pH = 4.9, B:
acetonitrile: water [50 : 50]) at a flow rate of 1 mL/min.
4. A diode array detector.
5. An UV absorbance at 254 nm.
6. Injection volume: 20 μl

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