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A validated new method for nevirapine quantitation in human plasma via high‐performance liquid chromatography
Author(s) -
Silverthorn Courtney F.,
Parsons Teresa L.
Publication year - 2006
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.519
Subject(s) - chromatography , chemistry , nevirapine , high performance liquid chromatography , detection limit , extraction (chemistry) , quantitative analysis (chemistry) , sample preparation , human immunodeficiency virus (hiv) , medicine , family medicine , viral load , antiretroviral therapy
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 C 8 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[sol ]mL ( r 2 > 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[sol ]mL for 50 µL 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. Copyright © 2005 John Wiley & Sons, Ltd.