High-performance liquid chromatographic method for the simultaneous determination of the six HIV-protease inhibitors and two non-nucleoside reverse transcriptase inhibitors in human plasma.
A selective and sensitive high-performance liquid chromatographic (HPLC) method has been developed for the determination of the six human immunodeficiency virus (HIV)-protease inhibitors (amprenavir, indinavir, lopinavir, nelfinavir, ritonavir, and saquinavir) and the non-nucleoside reverse transcriptase inhibitors (efavirenz and nevirapine) in a single run. After a liquid-liquid extraction with diethyl ether, the six protease inhibitors and the two non-nucleoside reverse transcriptase inhibitors are separated on a Stability RP18 column eluted with a gradient of acetonitrile and phosphate buffer 50 mmol/L pH 5.65. A sequential ultraviolet detection (5-minute sequence set at 240 nm for nevirapine acquisition, 22-minute sequence set at 215 nm for other antiretroviral drugs acquisition followed by a sequence set at 260 nm for internal standard acquisition) allowed for simultaneous quantitation of the six protease inhibitors, nevirapine, and efavirenz. Calibration curves were linear in the range 100 ng/mL to 10,000 ng/mL. The limit of quantitation was 50 ng/mL for all drugs except nevirapine (100 ng/mL). Average accuracy at four concentrations ranged from 88.2% to 110.9%. Both interday and intraday coefficients of variation were less than 11% for all drugs. The extraction recoveries were greater than 62%. This method is simple and shows a good specificity with respect to commonly co-prescribed drugs. This method allows accurate therapeutic monitoring of amprenavir, indinavir, lopinavir, nelfinavir, ritonavir, saquinavir, efavirenz, and nevirapine.
Titier
Karine
K
Department of Clinical Pharmacology and Toxicology, University Victor Ségalen, Pellegrin Hospital, Place Amélie Raba-Léon, 33076 Bordeaux cedex, France.
Lagrange
Fabrice
F
Péhourcq
Fabienne
F
Edno-Mcheik
Leila
L
Moore
Nicholas
N
Molimard
Mathieu
M
D000480:Alkynes; D048588:Benzoxazines; D002219:Carbamates; D002851:Chromatography, High Pressure Liquid; D003521:Cyclopropanes; D016903:Drug Monitoring / Q000379:methods / Q000592:standards*; D005663:Furans; D015658:HIV Infections / Q000188:drug therapy; D006801:Humans; D019469:Indinavir / Q000097:blood; D061466:Lopinavir; D019888:Nelfinavir / Q000097:blood; D019829:Nevirapine / Q000097:blood; D010078:Oxazines / Q000097:blood; D011480:Protease Inhibitors / Q000097:blood* / Q000627:therapeutic use; D011744:Pyrimidinones / Q000097:blood; D015203:Reproducibility of Results; D018894:Reverse Transcriptase Inhibitors / Q000097:blood* / Q000627:therapeutic use; D019438:Ritonavir / Q000097:blood; D019258:Saquinavir / Q000097:blood; D013449:Sulfonamides / Q000097:blood
D000480:Alkynes; D048588:Benzoxazines; D002219:Carbamates; D003521:Cyclopropanes; D005663:Furans; D010078:Oxazines; D011480:Protease Inhibitors; D011744:Pyrimidinones; D018894:Reverse Transcriptase Inhibitors; D013449:Sulfonamides; D061466:Lopinavir; C095108:amprenavir; D019469:Indinavir; D019829:Nevirapine; D019888:Nelfinavir; C098320:efavirenz; D019258:Saquinavir; D019438:Ritonavir