High-performance liquid chromatographic assay for pilocarpine in aqueous humor
1984; Elsevier BV; Volume: 336; Issue: 1 Linguagem: Inglês
10.1016/s0378-4347(00)85157-9
ISSN1872-812X
AutoresMaris A. Rendi, Philip P. Ellis, J. Gál,
Tópico(s)Toxin Mechanisms and Immunotoxins
ResumoA 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 100μl 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.
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