Ramesh Bokka, Rao Vadaparthi P Rajesh, Sukumar Genji, Manjula Nemali, Suresh Babu Katragadda, Sita Devi Potturi
Natural Products Chemistry Division, Indian Institute of Chemical Technology, Tarnaka, Hyderabad 500607, India.
J Pharm Anal. 2016 Feb;6(1):18-23. doi: 10.1016/j.jpha.2015.07.002. Epub 2015 Jul 20.
A liquid chromatography-high resolution mass spectrometry (LC-HRMS) method was developed and validated for the determination of piperine (PPR) on dried blood spots (DBS). DBS samples were prepared by spiking the whole blood with analyte to produce 30 µL of blood spots on specimen collection cards. Chromatographic separation was achieved on an Atlantis dC column using acetonitrile and water (0.1% formic acid) (85:15, v/v) as mobile phase in an isocratic mode of elution at a flow rate of 0.75 mL/min. MS detection was carried out in electrospray positive ion mode for the target ions and monitored at 286.1465 for PPR and 272.1303 for the internal standard (IS). The developed method exhibited a linear dynamic range over 0.01-2000 ng/mL for PPR on DBS. The overall extraction recovery of PPR from DBS was 92.5%. Influence of hematocrit and spot volume on DBS was also evaluated and found to be well within the acceptable limits. The method was successfully applied to pharmacokinetic studies of PPR in rats.
建立了一种液相色谱-高分辨率质谱（LC-HRMS）方法并进行了验证，用于测定干血斑（DBS）中的胡椒碱（PPR）。通过在全血中加入分析物来制备DBS样品，以在标本采集卡上产生30μL血斑。在Atlantis dC柱上进行色谱分离，使用乙腈和水（0.1%甲酸）（85:15，v/v）作为流动相，以等度洗脱模式，流速为0.75 mL/min。MS检测在电喷雾正离子模式下对目标离子进行，PPR监测质荷比为286.1465，内标（IS）监测质荷比为272.1303。所建立的方法在DBS上对PPR的线性动态范围为0.01 - 2000 ng/mL。从DBS中提取PPR的总体回收率为92.5%。还评估了血细胞比容和血斑体积对DBS的影响，发现其均在可接受范围内。该方法成功应用于大鼠中PPR的药代动力学研究。
