Clark T Nicole, White Catherine A, Bartlett Michael G
Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, The University of Georgia, Athens, 30602-2352, USA.
Biomed Chromatogr. 2006 Jun-Jul;20(6-7):605-11. doi: 10.1002/bmc.651.
A rapid and efficient high-performance liquid chromatography (HPLC)-tandem mass spectrometry method for the determination of didanosine concentrations in maternal rat plasma, amniotic fluid, placental and fetal tissue samples has been developed and validated. Tissue samples were homogenized in optima water and centrifuged. The supernatant was subjected to solid-phase extraction (SPE) prior to analysis. Plasma and amniotic fluid samples were extracted without pretreatment. An Agilent 1100 Series HPLC coupled with a Micromass Quattro II triple quadrupole mass spectrometer was used for all analyses. Chromatographic resolution was achieved on a Nova-Pak phenyl analytical column (2.0 x 150 mm, 4 microm particle size) equipped with a Phenomenex Security-guard phenyl guard cartridge (2.0 x 4.0 mm) using 60% methanol in 10 mm ammonium acetate buffer mobile phase for all matrices at a flow rate of 0.15 mL/min. The method yields retention times of 2.9 min for didanosine and 3.0 min for the internal standard, stavudine. Limits of detection were 1 ng/mL for all matrices. Recoveries were 70% or greater for both compounds in the different matrices. Within- and between-run precision (%RSD) and accuracy (%error) was less than 15% for all matrices.
已开发并验证了一种快速高效的高效液相色谱（HPLC）-串联质谱法，用于测定孕鼠血浆、羊水、胎盘和胎儿组织样本中的去羟肌苷浓度。将组织样本在超纯水中匀浆并离心。上清液在分析前进行固相萃取（SPE）。血浆和羊水样本无需预处理即可进行萃取。所有分析均使用安捷伦1100系列HPLC与Micromass Quattro II三重四极杆质谱仪联用。在配备Phenomenex Security-guard苯基保护柱（2.0×4.0 mm）的Nova-Pak苯基分析柱（2.0×150 mm，4μm粒径）上实现色谱分离，所有基质均使用含60%甲醇的10 mM醋酸铵缓冲液作为流动相，流速为0.15 mL/min。该方法测得去羟肌苷的保留时间为2.9分钟，内标司他夫定的保留时间为3.0分钟。所有基质的检测限均为1 ng/mL。两种化合物在不同基质中的回收率均达到70%或更高。所有基质的批内和批间精密度（%RSD）和准确度（%误差）均小于15%。
