高速液体クロマトグラフィー並びにカラムクロマトグラフィーによるアスコルビン酸2-硫酸の定量法

概要

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Prior to investigation of metabolism of ascorbic acid 2-sulfate (AsS) in guinea pig, methods to determine the amount of AsS in serum, urine, liver and other organs intubated with AsS have been studied by high speed liquid chromatography (HSLC) and column chromatography (CPC). AsS in organs was extracted with water by using homogenizer, and the resulting extracts were filtered with a filter (0.45μ) after centrifugation 100,000 x g, 30 minutes. 1) Method for HSLC : the operating conditions are following, instrument; Model JASCO HSLC 350,detector; UV photometer 254nm, column; JASCOPAC AV 02-500,mobilephase; 0.05M KH_2PO_4 : 0.5M KH_2PC_4=7 : 3 v/v, column pressure; 50kg/cm^2,flow rate; 1.0ml/min, sample volume; 2〜2Oμl and retention time; 10 minutes. 2) Method for CPC : chromatography of extracts 0.1〜2ml were performed with a column (0.6×2cm) of DEAE-Sephadex A-25(C1-type), column were washed with 50ml of water and 50ml of 0.1M KH_2PO_4 after absorbing, AsS eluted with 1M KH_2PO_4. The elution was followed by measurement of ultraviolet absorbancy at 254nm. The lower limit of detection were following, HSLC : serum 0.05pmol/m1,urine 0.2-=0.3μmol/m1,other organs 0.5μmol/g. CPC : serum 0.05μmol/m1,urine 0.2〜0.3μmol/m1,other organs 0.8μmol/g.
日本ビタミン学会の論文
1977-06-25