The Microdetermination of Ascorbic acid in Blood by Gas-Liquid Chromatography

혈액중(血液中) Ascorbic acid 의 Gas-Liquid Chromatography 에 의(依)한 미량정량(微量定量)

A microdetermination of Ascorbic acid (AsA) in blood by Gas-Liquid Chromatography (GLC) was studied. AsA was applied on GLC after the conversion into trimethylsilyl derivative (TMS) and the GLC was available for the only reduced form of AsA. A calibration curve is made by GLC of TMS-AsA as the internal standard of n-docosane. The minimum amount of AsA required for the determination was 0.5ml of 1 mg% pyridine solution. Prior to the conversion of AsA in serum into TMS derivative, serum was lyophilized and then it was allowed to stand at room temperature with TMS reagents for 48 hr. On injection of the supernates of TMS derivative to GLC the peak corresponding to AsA was not detected. Its reason why the concentration of AsA in serum is $0.5mg{\sim}0.8mg%$ in general, and it is less than minimum concentration of GLC. In case of L-AsA 1 mg was added to 1ml of serum, which was followed by lyophilization, silylation and GLC. The recorvery of AsA added was 98 percentage.

1) 본실험조건(本實驗條件)에서 GLC에 의하여 AsA의 최저(最低)농도(濃度) 1mg%까지 정량가능(定量可能)하였다. 2) TMS 반응조건(反應條件)으로서 동결건조시료(凍結乾燥試料)에는 반드시 TMS 혼합시액중(混合試液中)에서 추출(抽出)과 TMS 화(化)를 동시(同時)에 시켜야만 98% 이상(以上)의 회수율(回收率)을 얻을 수 있다. 3) AsA 농도(濃度)가 1mg%이상(以上)인 시료(試料)에 대(對)하여 GLC에 의한 미량정량(微量定豊)을 확립(確立)하여 특(特)히 방해물질(妨害物質)이 많은 일반식품(一般食品)의 정량(定量)에 적용(適用)이 가능(可能)하도록 하였다. 4) 본실험(本實驗)에 사용(便用)한 일본전자(日本電子)의 750형(型)의 eletron meter의 성능(性能)보다 감도(感度)가 300배(倍)인 20K 형(型)을 사용(使用) 한다면 혈청중(血淸中) AsA의 정량(定量) 및 그 이하(以下)의 농도(濃度)인 미량정량(微量定量)도 충분(充分)히 가능(可能)하리라 생각된다.