采用柱切换直接进样液相色谱法测定人尿中新型唾液酸类似物GG167（GR121167X）

Determination of the novel sialic acid analog GG167 (GR121167X) in human urine by liquid chromatography: direct injection with column switching.

作者信息

Morris D M, Hussey B K, Geurin S L, Selinger K A

机构信息

Department of Clinical Pharmacology, Glaxo Inc. Research Institute, Research Triangle Park, NC 27709, USA.

出版信息

J Pharm Biomed Anal. 1995 Dec;14(1-2):191-201. doi: 10.1016/0731-7085(95)01626-0.

DOI:10.1016/0731-7085(95)01626-0

PMID:8833982

Abstract

GG167 is a novel compound which selectively inhibits viral neuraminidase and has demonstrated activity against influenza A and B. A liquid chromatography (LC) method for the determination of GG167 in human urine has been developed and validated. The method allows direct injection of urine (7 microliters) using LC column switching followed by UV detection. Initial chromatography is performed using a Nucleosil-Diol column (7 microns, 250 mm x 4.6 mm), eluted with 20 mM phosphate buffer (pH 2.5):acetonitrile (18:82, v/v) at 2.0 ml min-1. GG167 is "heart-cut" to a Spherisorb-SCX column (5 microns, 100 mm x 4.6 mm) and eluted with 35 mM phosphate buffer (pH 2.5):acetonitrile (50:50, v/v) at 1.5 ml min-1 for final separation. GG167 is detected by UV absorbance at lambda = 238 nm. UV detection and peak shape are enhanced at pH < 2.5. The quantitation range of the assay is 0.3-100 micrograms ml-1. The method has demonstrated sufficient ruggedness to be used in support of GG167 clinical trials.

摘要

GG167是一种新型化合物，它能选择性抑制病毒神经氨酸酶，并已证明对甲型和乙型流感有活性。已开发并验证了一种用于测定人尿中GG167的液相色谱（LC）方法。该方法采用LC柱切换后直接进样尿液（7微升），随后进行紫外检测。初始色谱分析使用Nucleosil - Diol柱（7微米，250毫米×4.6毫米），以20 mM磷酸盐缓冲液（pH 2.5）：乙腈（18:82，v/v）在2.0毫升/分钟的流速下洗脱。GG167被“中心切割”到Spherisorb - SCX柱（5微米，100毫米×4.6毫米），并以35 mM磷酸盐缓冲液（pH 2.5）：乙腈（50:50，v/v）在1.5毫升/分钟的流速下洗脱进行最终分离。通过在λ = 238纳米处的紫外吸光度检测GG167。在pH < 2.5时，紫外检测和峰形得到改善。该测定法的定量范围为0.3 - 100微克/毫升。该方法已证明具有足够的耐用性，可用于支持GG167的临床试验。