Silverthorn Courtney F, Parsons Teresa L
Division of Clinical Pharmacology, Department of Medicine, The Johns Hopkins University School of Medicine, 600 North Wolfe Street, Osler 527, Baltimore, MD 21287-5554, USA.
Biomed Chromatogr. 2006 Jan;20(1):23-7. doi: 10.1002/bmc.519.
A fully validated and clinically relevant assay was developed for the assessment of nevirapine concentrations in neonate blood plasma samples. Solid-phase extraction with an acid-base wash series was used to prepare subject samples for analysis. Samples were separated by high performance liquid chromatography and detected at 280 nm on a C8 reverse-phase column in an isocratic mobile phase. The retention times of nevirapine and its internal standard were 5.0 and 6.9 min, respectively. The method was validated by assessment of accuracy and precision (statistical values <15%), specificity, and stability. The assay was linear in the range 25-10,000 ng/mL (r2 > 0.996) and the average recovery was 93% (n = 18). The lower limit of quantification (relative standard deviation <20%) was determined to be 25 ng/mL for 50 microL of plasma, allowing detection of as little as 1.25 ng of nevirapine in a sample. This value represents an increase in sensitivity of up to 30-fold over previously published methods.
已开发出一种经过充分验证且具有临床相关性的检测方法，用于评估新生儿血浆样本中的奈韦拉平浓度。采用酸碱洗涤系列的固相萃取法制备待分析的受试者样本。通过高效液相色谱法分离样本，并在等度流动相中的C8反相柱上于280 nm处进行检测。奈韦拉平及其内标的保留时间分别为5.0分钟和6.9分钟。通过评估准确度和精密度（统计值<15%）、特异性和稳定性对该方法进行了验证。该检测方法在25 - 10,000 ng/mL范围内呈线性（r2 > 0.996），平均回收率为93%（n = 18）。对于50微升血浆，定量下限（相对标准偏差<20%）确定为25 ng/mL，这使得能够检测出样本中低至1.25 ng的奈韦拉平。该值表示灵敏度比先前发表的方法提高了高达30倍。
