Pantothenic Acid. Shang-Jing Pan, Ph.D. Abbott Nutrition Columbus, Ohio, USA

Transcription

1 Pantothenic Acid Shang-Jing Pan, Ph.D. Abbott Nutrition Columbus, Ohio, USA

2 Pantothenic acid- history Discovered by Roger J Williams in Isolated in 1933 by R. J. Williams. Named by Williams, meaning from everywhere for its widespread occurrence. Total synthesis was first achieved by the US Merck in 1940.

3 Pantothenic acid- background Also called vitamin B 5. Principal biological active form: coenzyme A (CoA). Second active form: 4 phosphopanthetheine moiety in acyl-carrier protein (ACP). Sodium and calcium salt are often used to fortify food and feed. Pantothenol is usually used in liquid pharmaceutical preparations and in cosmetics.

4 Pantothenic acid- structure Pantothenic acid

5 Pantothenic acid- structure Calcium pantothenate

6 pantothenol Pantothenic acid- structure

7 Pantothenic acid- vitamin activity CoA is integral in the metabolism of carbohydrates, lipids, and nitrogencontaining compounds, in TCA cycle, in the synthesis of fatty acids, phospholipids, heme, and steroids, and in regulating many proteins via acetylation or acylation. ACP participates in the biosynthesis of long-chain fatty acids.

8 Pantothenic acid- vitamin activity No IU has been defined. Analytical results generally reported in weight units of pantothenic acid. 1 mg pantothenic acid = mg of calcium pantothenate. Only the d- isomer is biologically active.

9 Pantothenic acid- analytical method in Foods Indirect measurement Microbiological method Biosensor Direct measurement Chromatography HPLC, UPLC, GC, UV, fluorescence, FTIR, Mass spec.

10 Pantothenic acid- analytical method in Foods Microbiological assay AOAC official method Applicable to milk-based RTF infant formula Employs enzyme digestion to quantitate total AOAC official method For vitamin preparations Issues: time, materials (maintaining test organism culture), precision. Alternative: microtiter plate format with mircoorganisms coated on the plate.

11 Pantothenic acid- analytical method in Foods Biosensor method AOAC certified, performance tested status (cert. #090601) Optical biosensor technique, measures difference in surface plasmon resonance of immobilized ligand before and after sample introduction affinity-based competitive inhibition protein binding assay Free pantothenic acid Published report showing data supporting applicability to various food products and NIST SRMs, assay sample LOQ18.9 ng/ml.

12 Pantothenic acid- analytical method in Foods Chromatography methods PA, low volatility for direct GC, need derivatization LC, direct analysis, sensitive and specific, could be used for simultaneous determination of several vitamins. PA, low UV absorption, derivatization required for fluorometric detection. UPLC tandem MS

13 Pantothenic acid- analytical method in Foods HPLC methods- UV detection Reversed phase C18 Low ph (2.5) mobile phase UV detection (195 nm, 197 nm) Published reports showing data supporting applicability to milk-based formula, LOQ 1 mg/100g

14 Pantothenic acid- analytical method in Foods HPLC methods- fluorometric detection Reversed phase C18, Low ph (2.5) mobile phase Ion exchange SPE pre-column purification Post column derivatization for fluorometric detection Published reports showing data supporting applicability to various foods, LOQ 65 mcg/100g

15 Pantothenic acid- analytical method in Foods LC-MS/MS Reversed phase C18 Stable isotope internal standard Tandam mass spec with positive-ion electrospray ionization (ESI+) Simultaneous determination of several vitamins Published reports showing data supporting applicability to various foods, LOQ 24 mcg/100g

16 Pantothenic acid- regulation mcg/100 Kcal min max/gul Ref US, IF 300 NS IFA:21 CFR EU, IF & toddler /141/EC EU, FSMP-IF /21/EC Codex, IF Codex stan Codex, FSMP-IF Codex stan Codex toddler 300 NS Codex stan Australia/NZ, IF Standard 2.9.1

17 Pantothenic acid- US DRI DRI (mg/day) 0 through 6M through 12M through 3yr 2 4 through 8yr 3 9 through 13yr 4 14yr and older 5 Pregnancy 6 lactation 7

18 Pantothenic acidproposed fitness for purpose statement An analytical method to determine the fortified level of d- pantothenic acid (and pantothenate salts) in infant/pediatric/ adult nutritional products. These products could be made from any combination of milk, soy, rice, hydrolyzed proteins and amino acids without intact proteins. LOQ of the method should be lower than 300 mcg/100g. Result should be calculated to weight units in pantothenic acid. Method should be verified by NIST SRM The intended use of this method is as a reference method for dispute resolution.