Validation of HPLC Methods for Ascorbic Acid and Its Derivatives in Foods

Analytical methods for food antioxidants including ascorbic acid, erythorbic acid, ascorbyl palmitate (AP), and ascorbyl stearate (AS), were validated using high performance liquid chromatography. Validation parameters such as linearity, limit of detection (LOD), limit of quantification (LOQ), and recovery were tested using lard and cider as food model systems. Linearity of ascorbic acid and erythorbic acid were both higher than ($R^2$> 0.99), LOD of these compounds were 0.46 and 0.48 ${\mu}g/mL$, respectively and LOQ were 1.39 and 1.45 ${\mu}g/mL$, respectively. The recovery rates of these compounds were 86.35-94.78% and 84.76-95.02%, respectively. However, the concentration of AP and AS decreased in methanol stock solution. Four other solvents including ethanol, acetonitrile, mixture of methanol and acetonitrile, and mixture of ethanol and acetonitrile were tested to increase the stability of AP and AS under room temperature and refrigerated temperature. Ethanol provided better stability of AP and AS under both room and refrigerated temperature. This study can help to accurately analyze the content of ascorbic acid and its derivatives in processed foods.

식품 중 아스코르빈산 유래 산화방지제의 HPLC 분석법 검증 및 개선

산화방지제인 아스코르빈산(AA), 에리쏘르빈산(isoAA), 아스코르빌파르미테이트(AP), 아스코르빌스테아레이트(AS)에 대한 '식품 중 식품첨가물 분석법'을 검증하고 미비점을 개선하였다. 고속액체크로마토그래피 자외선검출기법을 이용하여 검출한계(LOD), 정량한계(LOQ), 상관계수($R^2$) 등을 측정하였고 돼지기름(lard), 사이다를 이용한 모델식품에서의 회수율과 재현성을 측정하였다. AA와 isoAA의 검출한계는 각각 0.46, 0.48 ${\mu}g/mL$이었으며, 회수율은 각각 86.35-94.78, 84.76-95.02%를 나타내었고 상관계수는 모두 0.999이상을 보였다. AP와 AS의 현 분석법은 메탄올을 용매로 사용하지만 메탄올 용매에서 AP와 AS는 불안정하였다. 냉장온도에서 에탄올을 용매로 사용 시 다른 용매에 비해 유의적으로 높은 안정성을 나타내어 기존 용매의 불안정성을 개선할 수 있었다.