Gomes Noel A, Laud Avdhoot, Pudage Ashutosh, Joshi Santosh S, Vaidya Vikas V, Tandel Jayram A
Department of Chemistry, Ramnarain Ruia College, Mumbai University, Matunga, Mumbai, 400 019, India.
J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Jan 15;877(3):197-206. doi: 10.1016/j.jchromb.2008.12.013. Epub 2008 Dec 11.
The present research work involves a first of its kind rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method developed and validated for simultaneous analysis of Alverine (ALV) and one of its hydroxy metabolites, para hydroxy Alverine (PHA) in human plasma. The analytes were extracted from the matrix using a simple solid-phase extraction procedure. Mebeverine was used as the internal standard for both analytes. A Kromasil C8 column provided chromatographic separation of analytes followed by detection with mass spectrometry. The method involves simple isocratic chromatography conditions and mass spectrometric detection in the positive ionization mode using an API 5000 MS/MS system. The proposed method has been validated with a linear range of 100-10,000 pg/mL for both ALV and PHA. The interrun and intrarun precision values are within 6.3%, 3.7% for ALV and 6.3%, 3.2% for PHA at LOQ levels. The intrarun accuracy in terms of % RE was within the range of -7.0% to -0.1% and -8.1% to -1.7% for ALV and PHA, respectively whereas the interrun accuracy was within the range of -5.1% to -0.5% for ALV and -8.6% to 0.4% for PHA, respectively. The overall recoveries for ALV and PHA were 83.5% and 86.2% respectively. Total elution time was about 4 min which allowed quantitation of more than 150 plasma samples per day. This validated method was used successfully for analysis of real samples from a bioequivalence study.
本研究工作涉及一种首次开发并验证的快速灵敏的液相色谱 - 串联质谱（LC-MS/MS）方法，用于同时分析人血浆中的阿尔维林（ALV）及其一种羟基代谢物对羟基阿尔维林（PHA）。使用简单的固相萃取程序从基质中提取分析物。美贝维林用作两种分析物的内标。采用Kromasil C8柱对分析物进行色谱分离，随后用质谱进行检测。该方法采用简单的等度色谱条件，并使用API 5000 MS/MS系统在正离子模式下进行质谱检测。所提出的方法已得到验证，ALV和PHA的线性范围均为100 - 10,000 pg/mL。在定量限水平下，ALV的批间和批内精密度值分别在6.3%、3.7%以内，PHA的批间和批内精密度值分别在6.3%、3.2%以内。ALV的批内准确度（以%相对误差计）在 - 7.0%至 - 0.1%范围内，PHA的批内准确度在 - 8.1%至 - 1.7%范围内，而ALV的批间准确度在 - 5.1%至 - 0.5%范围内，PHA的批间准确度在 - 8.6%至0.4%范围内。ALV和PHA的总回收率分别为83.5%和86.2%。总洗脱时间约为4分钟，这使得每天能够定量分析超过150份血浆样品。这种经过验证的方法已成功用于生物等效性研究中实际样品的分析。
