Gal J, Ellis P P, Rendi M
Curr Eye Res. 1981;1(6):361-5. doi: 10.3109/02713688108998363.
A high-performance liquid chromatographic (HPLC) assay for acetazolamide is presented. A 100-microliter sample is mixed with an aliquot of the internal standard solution and the mixture, buffered at pH 4.5, is extracted with ethyl acetate. The extract is evaporated to dryness and the residue is analyzed by HPLC, using a reverse-phase octadecylsilane column. The wavelength of the detection is 254 nm. The coefficient of variation (CV) in the within-day analysis of replicate 10-microgram/ml acetazolamide samples in human blood plasma was 6.5%, while the between-day CV was 7.1%. The procedures was developed for the 1-25 microgram/ml acetazolamide concentration range. The internal standard used is similar in chemical structure to acetazolamide and can be readily prepared in one step from a commercially available precursor. In addition to blood serum or plasma, the assay can also use aqueous and vitreous humor samples. Theophylline and acetaminophen interfere in the assay. The technique was used to determine the concentration of acetazolamide in the blood serum of human volunteers after an oral dose of the drug, and in the aqueous and vitreous humors of rabbits after an intravenous dose of acetazolamide.
本文介绍了一种用于乙酰唑胺的高效液相色谱（HPLC）测定法。将100微升样品与一份内标溶液混合，在pH 4.5缓冲的混合物用乙酸乙酯萃取。提取物蒸发至干，残留物用反相十八烷基硅烷柱通过HPLC分析。检测波长为254nm。人血浆中10微克/毫升乙酰唑胺重复样品日内分析的变异系数（CV）为6.5%，日间CV为7.1%。该方法适用于1 - 25微克/毫升的乙酰唑胺浓度范围。所用内标在化学结构上与乙酰唑胺相似，可从市售前体一步轻松制备。除血清或血浆外，该测定法还可使用房水和玻璃体液样品。茶碱和对乙酰氨基酚会干扰该测定法。该技术用于测定口服药物后人类志愿者血清中乙酰唑胺的浓度，以及静脉注射乙酰唑胺后兔房水和玻璃体液中乙酰唑胺的浓度。
