Quantitative Analysis of HIV-1 Protease Inhibitors in Cell Lysates Using MALDI-FTICR Mass Spectrometry
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- 作者：Jeroen J. A. van Kampen ; Peter C. Burgers ; Ronald de Groot ; Albert D. M. E. Osterhaus ; Mariska L. Reedijk ; Esther J. Verschuren ; Rob A. Gruters ; Theo M. Luider
- 刊名：Analytical Chemistry
- 出版年：2008
- 出版时间：May 15, 2008
- 年：2008
- 卷：80
- 期：10
- 页码：3751 - 3756
- 全文大小：140K
- 年卷期：v.80,no.10(May 15, 2008)
- ISSN：1520-6882
文摘
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/µL for saquinavir, 16 fmol/µL for lopinavir, 31 fmol/µL for ritonavir, and 100 fmol/µL 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.