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已有的 HPLC 测定生物液体中氢氯噻嗪(Ⅰ)浓度的方法大多需繁琐的样品处理步骤,采用本文方法,血浆样品可直接注入色谱系统作全自动分析。溶液的制备分别配制1.0mg/ml 的(Ⅰ)和内标氢氟噻嗪(Ⅱ)的甲醇溶液作为贮备液,于-20℃保存。将(Ⅰ)的贮备液用10mmol/L 的磷酸盐缓冲液(pH7.2)稀释至100μg/ml,并以空白血浆(先离心去微粒)进一步稀释制得血浆标准液。以相同缓冲液稀释内标液至1500ng/ml。血浆样品室温解冻后离心(3000rpm,10min)去微粒,将150μl 血浆标准液或样品液与150μl 内标液混合,采用自动进样器将100～200μl 混合液注入 HPLC 仪。色谱系统包括提取柱(两个串联的Guard-Pak C_(18)柱,柱长0.4cm)和串联Guard-Pak C_(18)保护柱的分析柱(HypersilODS,5μm,4.6×250mm),分别联于六孔
Existing HPLC methods for determining the concentration of hydrochlorothiazide (I) in biological fluids often require cumbersome sample processing steps. Using this method, plasma samples can be injected directly into the chromatography system for automated analysis. Solution Preparation A methanol solution of 1.0 mg / ml (I) and an internal standard hydroflumethiazine (II) was prepared as a stock solution and stored at -20 ° C. The stock solution of (I) was diluted to 100 μg / ml with 10 mmol / L phosphate buffer (pH 7.2) and further diluted with blank plasma (centrifuge to remove particulates) to prepare a plasma standard solution. The internal standard solution was diluted to 1500 ng / ml with the same buffer. Plasma samples were thawed at room temperature and centrifuged (3000 rpm, 10 min) to remove particulates. 150 μl of the plasma standard or sample solution was mixed with 150 μl of internal standard solution and 100-200 μl of the mixture was injected into the HPLC instrument using an autosampler. The chromatographic system consisted of an analytical column (Hypersil ODS, 5 μm, 4.6 × 250 mm) equipped with an extraction column (two Guard-Pak C_ (18) columns in series with a column length of 0.4 cm) and a Guard- Pak C_ In six holes