Matsuura K, Kuwano M, Takashina H
Central Research Laboratories, Santen Pharmaceutical Co., Osaka, Japan.
J Chromatogr. 1993 Nov 24;621(2):173-80. doi: 10.1016/0378-4347(93)80093-j.
A new method has been developed for rapid analysis and determination of pilocarpine in aqueous humour using liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. The chromatography was carried out on a reversed-phase phenyl column with 0.1% acetic acid-acetonitrile (95:5, v/v). Pilocarpine and its analogues, isopilocarpine, pilocarpic acid and isopilocarpic acid, were separated. An aqueous humour sample was deproteinized with methanol. After evaporation, the residue was dissolved in the mobile phase. The method was applied to the analysis of the metabolite in aqueous humour after the topical application of 2% pilocarpine (w/v) eye-drops. The main metabolite, pilocarpic acid, was easily identified. The protonated molecular ion of pilocarpine was used for the determination. The calibration curve had a good linearity within the concentration range investigated (2 ng to 10 micrograms/ml). The limit of determination was estimated to be an aqueous humour concentration of ca. 2 ng/ml. The method was applied to the determination of unchanged pilocarpine after the topical application of 2% pilocarpine (w/v) eye-drops.
已开发出一种新方法，用于使用液相色谱 - 大气压化学电离质谱法快速分析和测定房水中的毛果芸香碱。色谱分析在反相苯基柱上进行，流动相为0.1%乙酸 - 乙腈（95:5，v/v）。毛果芸香碱及其类似物异毛果芸香碱、毛果芸香酸和异毛果芸香酸得以分离。房水样品用甲醇进行脱蛋白处理。蒸发后，残留物溶解于流动相中。该方法应用于局部滴用2%（w/v）毛果芸香碱滴眼液后房水中代谢物的分析。主要代谢物毛果芸香酸很容易被鉴定出来。使用毛果芸香碱的质子化分子离子进行测定。在所研究的浓度范围（2 ng至10 μg/ml）内，校准曲线具有良好的线性。测定限估计为房水浓度约2 ng/ml。该方法应用于局部滴用2%（w/v）毛果芸香碱滴眼液后未变化的毛果芸香碱的测定。
