Quantitative analysis of HIV-1 protease inhibitors in cell lysates using MALDI-FTICR mass spectrometry
| Autor: | Jeroen J. A. van Kampen, Esther J. Verschuren, Theo M. Luider, Mariska L. Reedijk, Rob A. Gruters, Albert D. M. E. Osterhaus, Peter C. Burgers, Ronald de Groot |
|---|---|
| Přispěvatelé: | Neurology, Pediatrics, Virology |
| Rok vydání: | 2008 |
| Předmět: |
viruses
Sensitivity and Specificity Auto-immunity transplantation and immunotherapy [N4i 4] Analytical Chemistry Matrix (chemical analysis) SDG 3 - Good Health and Well-being Indinavir immune system diseases Spectroscopy Fourier Transform Infrared medicine Protease inhibitor (pharmacology) Cells Cultured Chromatography Chemistry Poverty-related infectious diseases [N4i 3] virus diseases Lopinavir HIV Protease Inhibitors biochemical phenomena metabolism and nutrition Pathogenesis and modulation of inflammation [N4i 1] Nelfinavir Biochemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Ritonavir Microbial pathogenesis and host defense [UMCN 4.1] Saquinavir Quantitative analysis (chemistry) medicine.drug |
| Zdroj: | Analytica Chimica Acta, 80, 10, pp. 3751-6 Analytica Chimica Acta, 80, 3751-6 Analytical Chemistry, 80(10), 3751-3756. American Chemical Society |
| ISSN: | 0003-2670 0003-2700 |
| DOI: | 10.1021/ac702072c |
| Popis: | Contains fulltext : 71284.pdf (Publisher’s version ) (Open Access) In this report we explore the use of MALDI-FTICR mass spectrometry for the quantitative analysis of five HIV-1 protease inhibitors in cell lysates. 2,5-Dihydroxybenzoic acid (DHB) was used as the matrix. From a quantitative perspective, DHB is usually a poor matrix due to its poor shot-to-shot and poor spot-to-spot reproducibilities. We found that the quantitative precisions improved significantly when DMSO (dimethylsulfoxide) was added to the matrix solution. For lopinavir and ritonavir, currently the most frequently prescribed HIV-1 protease inhibitors, the signal-to-noise ratios improved significantly when potassium iodide was added to the matrix solution. The mean quantitative precisions, expressed as % relative standard deviation, were 6.4% for saquinavir, 7.3% for lopinavir, 8.5% for ritonavir, 11.1% for indinavir, and 7.2% for nelfinavir. The mean quantitative accuracies, expressed as % deviation, were 4.5% for saquinavir, 6.0% for lopinavir, 5.9% for ritonavir, 6.6% for indinavir, and 8.0% for nelfinavir. The concentrations measured for the individual quality control samples were all within 85-117% of the theoretical concentrations. The lower limits of quantification in cell lysates were 4 fmol/microL for saquinavir, 16 fmol/microL for lopinavir, 31 fmol/microL for ritonavir, and 100 fmol/microL for indinavir and nelfinavir. The mean mass accuracies for the protease inhibitors were 0.28 ppm using external calibration. Our results show that MALDI-FTICR mass spectrometry can be successfully used for precise, accurate, and selective quantitative analyses of HIV-1 protease inhibitors in cell lysates. In addition, the lower limits of quantification obtained allow clinical applications of the technique. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect