Rendi M A, Ellis P P, Gal J
J Chromatogr. 1984 Dec 12;336(2):337-44. doi: 10.1016/s0378-4347(00)85157-9.
A high-performance liquid chromatographic assay for pilocarpine has been developed for the determination of pilocarpine in aqueous humor. A structurally similar internal standard is used, and pilocarpine is separated from isopilocarpine under the chromatographic conditions used. A 100-microliter sample is mixed with an aliquot of internal standard at pH 8.3 and extracted with methylene chloride. The extract is evaporated to dryness and the alkaloids are quaternized with p-nitrobenzyl bromide. Following the quaternization, the sample is evaporated to dryness, washed and diluted with a mobile phase--triethylamine mixture and analyzed by high-performance liquid chromatography using a reversed-phase octadecylsilane column with detection at a wavelength of 254 nm. This is a highly sensitive, reproducible and selective assay for measuring pilocarpine at physiological levels in individual aqueous humor samples.
已开发出一种用于测定房水中毛果芸香碱的高效液相色谱分析法。使用了一种结构相似的内标物，在所采用的色谱条件下，毛果芸香碱与异毛果芸香碱得以分离。将100微升样品与pH值为8.3的内标物等分试样混合，并用二氯甲烷萃取。提取物蒸发至干，生物碱用对硝基苄基溴季铵化。季铵化后，样品蒸发至干，用流动相 - 三乙胺混合物洗涤并稀释，然后使用反相十八烷基硅烷柱通过高效液相色谱法进行分析，检测波长为254nm。这是一种用于测量单个房水样品中生理水平毛果芸香碱的高灵敏度、可重现且选择性高的分析方法。
