Olagunju Adeniyi, Bolaji Oluseye O, Amara Alieu, Waitt Catriona, Else Laura, Soyinka Julius, Adeagbo Babatunde, Adejuyigbe Ebunoluwa, Siccardi Marco, Back David, Owen Andrew, Khoo Saye
Department of Molecular and Clinical Pharmacology, University of Liverpool, 70 Pembroke Place, Liverpool L69 3GF, UK Faculty of Pharmacy, Obafemi Awolowo University, Ile-Ife, Nigeria.
Faculty of Pharmacy, Obafemi Awolowo University, Ile-Ife, Nigeria.
J Antimicrob Chemother. 2015 Feb;70(2):555-61. doi: 10.1093/jac/dku420. Epub 2014 Oct 17.
This manuscript describes the development, validation and clinical application of a novel method for the quantification of the antiretroviral drug efavirenz in dried breast milk spots using LC-MS.
Dried breast milk spots were prepared by spotting 30 μL of human breast milk on each circle of Whatman 903 Protein Saver cards. Chromatographic separation was achieved on a reverse-phase C18 column with 1 mM ammonium acetate in water/acetonitrile using a solvent gradient at a flow rate of 400 μL/min and detection was by TSQ Quantum Access triple quadrupole mass spectrometer equipped with a heated electrospray ionization source. The method was applied to characterize the breast milk pharmacokinetic profile of efavirenz in HIV-positive nursing mothers receiving regimens containing 600 mg of efavirenz once daily.
The assay was validated over the concentration range 50-7500 ng/mL. Accuracy ranged between 95.2% and 102.5% and precision ranged between 1.05% and 9.53%. The average recovery of efavirenz from dried breast milk spots was 106.4% and the matrix effect was 8.14%. Stability of efavirenz in dried breast milk spots and processed samples at room temperature, -40°C and -80°C was demonstrated. In the pharmacokinetic study, the mean (SD) AUC0-24, Cmax and Cmin of efavirenz in breast milk were 59,620 ng·h/mL (17,440), 4527 ng/mL (1767) and 1261 ng/mL (755.9), respectively. The mean (range) milk-to-plasma concentration ratio over the dosing interval was 0.78 (0.57-1.26).
The dried breast milk spot method is simple, robust, accurate and precise, and can be used in settings with limited resources.
本手稿描述了一种使用液相色谱-质谱法（LC-MS）定量干母乳斑中抗逆转录病毒药物依非韦伦的新方法的开发、验证及临床应用。
通过在Whatman 903蛋白质保存卡的每个圆圈上点样30μL人母乳来制备干母乳斑。在反相C18柱上进行色谱分离，流动相为含1 mM醋酸铵的水/乙腈溶液，采用溶剂梯度洗脱，流速为400μL/min，通过配备加热电喷雾电离源的TSQ Quantum Access三重四极杆质谱仪进行检测。该方法应用于对接受每日一次600 mg依非韦伦治疗方案的HIV阳性哺乳期母亲母乳中依非韦伦的药代动力学特征进行表征。
该测定法在50 - 7500 ng/mL浓度范围内得到验证。准确度在95.2%至102.5%之间，精密度在1.05%至9.53%之间。依非韦伦从干母乳斑中的平均回收率为106.4%，基质效应为8.14%。证明了依非韦伦在干母乳斑及处理后的样品在室温、-40°C和-80°C下的稳定性。在药代动力学研究中，母乳中依非韦伦的平均（标准差）AUC0 - 24、Cmax和Cmin分别为59,620 ng·h/mL（17,440）、4527 ng/mL（1767）和1261 ng/mL（755.9）。给药间隔期间的平均（范围）母乳与血浆浓度比为0.78（0.57 - 1.26）。
干母乳斑法简单、稳健、准确且精密，可用于资源有限的环境。
