Chapron D J, White L B
J Pharm Sci. 1984 Jul;73(7):985-9. doi: 10.1002/jps.2600730732.
A high-performance liquid chromatographic method for the determination of acetazolamide in whole blood, plasma, and urine was developed. Samples of biological fluids containing various concentrations of acetazolamide were spiked with the internal standard, sulfadiazine. Samples were then mixed with a 50% ammonium sulfamate solution. Whole blood samples were heated for 25 s in boiling water. All samples were extracted with ethyl acetate; a phosphate buffer (pH 8.0) was used to wash the extracts. Acetazolamide was back-extracted into a glycine buffer (pH 10.0), which was then washed with ether. Separation of acetazolamide and internal standard from other biological constituents was achieved on a 10-micron C18 reverse-phase column using an acetonitrile-methanol-acetate buffer (pH 4.0). The eluant was monitored at 254 nm. All calibration curves were linear, and the results from reproducibility studies were excellent. Application of the method to human pharmacokinetic studies was demonstrated.
建立了一种用于测定全血、血浆和尿液中乙酰唑胺的高效液相色谱法。将含有不同浓度乙酰唑胺的生物流体样品加入内标磺胺嘧啶。然后将样品与50%氨基磺酸铵溶液混合。全血样品在沸水中加热25秒。所有样品用乙酸乙酯萃取；用磷酸盐缓冲液（pH 8.0）洗涤萃取液。将乙酰唑胺反萃取到甘氨酸缓冲液（pH 10.0）中，然后用乙醚洗涤。使用乙腈-甲醇-乙酸盐缓冲液（pH 4.0）在10微米C18反相柱上实现乙酰唑胺和内标与其他生物成分的分离。在254nm处监测洗脱液。所有校准曲线均呈线性，重现性研究结果良好。证明了该方法在人体药代动力学研究中的应用。
