Knebel N G, Winkler M
Department of Biological Research Biochemistry, ASTA Medica AG, Frankfurt, Germany.
J Chromatogr B Biomed Sci Appl. 1997 Nov 21;702(1-2):119-29. doi: 10.1016/s0378-4347(97)00381-2.
An on-line HPLC assay with tandem mass spectrometric detection for the fast and sensitive determination of reproterol in human plasma was developed, utilising a methylated structural analogue as the internal standard. Automated solid-phase extraction of diluted plasma samples, based on 250 microl plasma aliquots, at pH 6.5, allowed a reliable quantification of reproterol down to 400 pg/ml. Injection of 100 microl of plasma extracts onto a 30 mm x 4.6 mm reversed-phase guard column provided retention times ranging from 20 to 30 s for reproterol and the internal standard. The standard curves were linear from 0.2 to 200 ng/ml using weighted linear regression analysis (1/y2). The inter-assay and intra-assay accuracies were -0.9% and +3.2%, exhibiting a precision (C.V.) of +/- 11% and +/- 9.3%, respectively. Up to 100 unknowns may be analysed each 24 h per analyst.
建立了一种采用串联质谱检测的在线高效液相色谱法，用于快速灵敏地测定人血浆中的瑞普特罗，使用甲基化结构类似物作为内标。基于250微升血浆等分试样，在pH 6.5条件下对稀释的血浆样品进行自动固相萃取，可可靠地定量低至400 pg/ml的瑞普特罗。将100微升血浆提取物注入30 mm×4.6 mm反相保护柱，瑞普特罗和内标的保留时间为20至30秒。使用加权线性回归分析(1/y2)，标准曲线在0.2至200 ng/ml范围内呈线性。批间和批内准确度分别为-0.9%和+3.2%，精密度(C.V.)分别为±11%和±9.3%。每位分析人员每24小时最多可分析100个未知样品。
