Bartels H, Bartels R
Borstel Research Center, Germany.
J Chromatogr B Biomed Appl. 1996 Nov 15;686(2):235-40. doi: 10.1016/s0378-4347(96)00223-x.
A simple HPLC method has been developed that allows the sensitive determination of rifabutin (RBT) in human serum using on-line concentration and column switching. After pretreatment of the serum with acetonitrile and centrifugation, the samples were applied to a concentration column (CC) (Zorbax CN). Washing with phosphate buffer-methanol removed most of the contaminating substances. Via a six-port valve the CC was switched to the analytical mode. RBT was separated on a Chromspher RP 8 column (acetonitrile-phosphate buffer pH 7.4/sodium chloride) and determined photometrically at 278 nm. The lower limit of quantification for 200 microliters serum precipitated with 200 microliters acetonitrile and after injection of 2 x 150 microliters was 33 micrograms/l and linearity was observed up to 27 mg/l. Different modes of sample application (single, repeated, and different injection volume portions), as well as washing time, cycle time and different CC materials were investigated.
已开发出一种简单的高效液相色谱法，该方法可使用在线浓缩和柱切换技术灵敏地测定人血清中的利福布汀（RBT）。血清经乙腈预处理并离心后，将样品注入浓缩柱（CC）（Zorbax CN）。用磷酸盐缓冲液 - 甲醇洗涤可去除大部分污染物。通过六通阀将CC切换至分析模式。RBT在Chromspher RP 8柱（乙腈 - pH 7.4的磷酸盐缓冲液/氯化钠）上分离，并在278 nm处进行光度测定。用200微升乙腈沉淀200微升血清，进样2×150微升后的定量下限为33微克/升，在高达27毫克/升的范围内观察到线性关系。研究了不同的进样模式（单次、重复和不同进样体积部分）以及洗涤时间、循环时间和不同的CC材料。
