태규리 (한국표준과학연구원)

최기환 (한국표준과학연구원)

김병주 (한국표준과학연구원)

태규리 (한국표준과학연구원)

이홍희 (한국표준과학연구원)

최기환 (한국표준과학연구원)

김병주 (한국표준과학연구원)

Pantothenic acid (vitamin B₅) is an essential nutrient for human health. Beef, a major source of pantothenic acid, present in both free and coenzyme A–bound forms. This study developed an enzymatic hydrolysis method to release the vitamin from coenzyme A for the determination of total pantothenic acid in beef. Quantification was performed by isotope dilution–liquid chromatography–tandem mass spectrometry (ID–LC–MS/MS) using [¹³C₃, ¹⁵N]–pantothenic acid as an internal standard. Prior to the assay, ultra centrifugal filter successfully eliminated pantothenic acid from the enzyme mixture to a level that does not interfere with measurement. For enzymatic hydrolysis, beef samples were treated with a phosphatase and pigeon liver pantetheinase in tris buffer (pH 8). After incubation, the hydrolysates were purified using a reversed-phase solid-phase extraction (SPE) cartridge. Post-column infusion ionization profiling showed that the reversed-phase SPE method efficiently removed co-eluted impurities during LC–MS/MS, compared to protein precipitation. The calculated matrix effect assessed by post-extraction addition was less than 5 %. After optimizing enzymatic hydrolysis conditions with ID–LC–MS/MS, the developed method will be applied for a value assignment of the certified reference material.

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