5. Vitamin Analysis.pdf

9/23/24 VITAMINS Low molecular weight biologically active organic...

9/23/24 VITAMINS Low molecular weight biologically active organic compounds Minor constituents but essential for an organism’s normal health and growth Relatively labile compounds VITAMIN CONTENT ANALYSIS FST 102 2 3 IMPORTANCE SAMPLE PREPARATION Low temperature storage Compliance of companies Storage of samples using amber bottles with the latest labelling Extraction from biological matrix regulations – Ascorbic acid – cold extraction with Ensuring declared labels are metaphosphoric acid or acetic acid correct for products with – B1 and B2 – boiling/autoclaving in acid claims followed by acid treatment – Niacin (B3) – autoclaving in acid (non- cereals) or base (cereals) – A, E, D – organic solvent extraction, saponification, and re-extraction 4 5 1  9/23/24 METHODS Experimental rats Cod-liver oil Observation after deprived with A. Biossay Method Vitamin D administration several days Used to determine the concentration of purity or biological activity of vitamins Rarely used clinically Common for Vit B12 and D analysis only Example: Line test for deficiency studies of Vit D 6 7 METHODS Sample containing B. Microbiological Assay Solution with known Vitamins Makes use of microorganisms amount of specific Estimates the concentration of the potency of an vitamin Extract specific Vit. antibiotic by measuring the area of inhibition compared to a standard on a specific mcg Plate the extract Plate the extract Limited only to niacin, cyanocobalamin, and pantothenic acid 8 9 2  9/23/24 Example: Vitamin A Analysis METHODS Sample (Proper sample C. Physicochemical Assay prep) Includes spectrophotometric, fluorometric, β-carotene extraction using chromatographic, enzymatic, immunological, and acetone:hexane radiometric methods Separation of pigments Chromatographic methods using high-performance liquid using solid-liquid β-carotene standard chromatography (HPLC) are most preferred chormatography solution HPLC is used as a reference technique to analyze any type of vitamin, both water soluble and fat-soluble Collection of extract vitamin, in one sample 10 11 3