LC-MS/MS Method for Determination of Colistin in Human Plasma: Validation and Stability Studies  

作　　者：Nada H. Binhashim Syed N. Alvi Muhammad M. Hammami Nada H. Binhashim;Syed N. Alvi;Muhammad M. Hammami(Clinical Studies and Empirical Ethics Department, King Faisal Specialist Hospital & Research Center, Riyadh, KSA;Alfaisal University College of Medicine, Riyadh, KSA)

机构地区：[1]Clinical Studies and Empirical Ethics Department, King Faisal Specialist Hospital & Research Center, Riyadh, KSA [2]Alfaisal University College of Medicine, Riyadh, KSA

出　　处：《International Journal of Analytical Mass Spectrometry and Chromatography》2021年第1期1-11,共11页分析质谱与色谱期刊（英文）

摘　　要：A simple and reliable high performance liquid chromatography tandem mass spectrometry (LC-MS/MS) assay for the determination of colistin A and colistin B in human plasma was developed and validated. Clarithromycin was used as an internal standard (IS). Plasma extraction was performed using C-18 cartridges and methanol containing 0.1% formic acid. Analysis was performed using Atlantis dC18 (2.1 × 100 mm, 3 μm) column at room temperature and a mobile phase of 0.2% formic acid in acetonitrile and water (50:50, v:v), delivered at a flow rate of 0.2 ml/minute. Eluent was detected in the positive ion mode using electrospray ionization at the following transitions of mass to charge (m/z): colistin A, 585.6 → 101.4;colistin B, 578.7 → 101.3;and IS, 748.6 → 158.4. No interference by components of blank plasma or commonly used drugs was observed. The relationship between colistin A colistin B concentrations and their corresponding peak height ratios to the IS was linear over the range of 0.05 - 10 μg/ml. Inter-day coefficient of variation and bias were, respectively, ≤11.5% and <span style="white-space:nowrap;">&minus;</span>3.0 to 6.0 for colistin A and ≤9.9 and <span style="white-space:nowrap;">&minus;</span>4.7 to 3.0 for colistin B. Mean extraction recovery of colistin A, colistin B, and the IS were 97%, 94%, and 97%, respectively. The method was applied to assess the stability of colistin A and colistin B in processed samples (24 hr. at room temperature, 48 hours at <span style="white-space:nowrap;">&minus;</span>20<span style="white-space:nowrap;">&#176;</span>C) and unprocessed samples (24 hr. at room temperature, 8 weeks at <span style="white-space:nowrap;">&minus;</span>20<span style="white-space:nowrap;">&#176;</span>C) and after three cycles of freeze and thaw found to be ≥87%.A simple and reliable high performance liquid chromatography tandem mass spectrometry (LC-MS/MS) assay for the determination of colistin A and colistin B in human plasma was developed and validated. Clarithromycin was used as an internal standard (IS). Plasma extraction was performed using C-18 cartridges and methanol containing 0.1% formic acid. Analysis was performed using Atlantis dC18 (2.1 × 100 mm, 3 μm) column at room temperature and a mobile phase of 0.2% formic acid in acetonitrile and water (50:50, v:v), delivered at a flow rate of 0.2 ml/minute. Eluent was detected in the positive ion mode using electrospray ionization at the following transitions of mass to charge (m/z): colistin A, 585.6 → 101.4;colistin B, 578.7 → 101.3;and IS, 748.6 → 158.4. No interference by components of blank plasma or commonly used drugs was observed. The relationship between colistin A colistin B concentrations and their corresponding peak height ratios to the IS was linear over the range of 0.05 - 10 μg/ml. Inter-day coefficient of variation and bias were, respectively, ≤11.5% and <span style="white-space:nowrap;">&minus;</span>3.0 to 6.0 for colistin A and ≤9.9 and <span style="white-space:nowrap;">&minus;</span>4.7 to 3.0 for colistin B. Mean extraction recovery of colistin A, colistin B, and the IS were 97%, 94%, and 97%, respectively. The method was applied to assess the stability of colistin A and colistin B in processed samples (24 hr. at room temperature, 48 hours at <span style="white-space:nowrap;">&minus;</span>20<span style="white-space:nowrap;">&#176;</span>C) and unprocessed samples (24 hr. at room temperature, 8 weeks at <span style="white-space:nowrap;">&minus;</span>20<span style="white-space:nowrap;">&#176;</span>C) and after three cycles of freeze and thaw found to be ≥87%.

关 键 词：COLISTIN CLARITHROMYCIN Human Plasma LC-MS/MS 

分 类 号：O62[理学—有机化学]