Mistri Hiren N, Jangid Arvind G, Sanyal Mallika, Shrivastav Pranav
Department of Chemistry, School of Sciences, Gujarat University, Navrangpura, Ahmedabad 380009, India.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 May 1;850(1-2):318-26. doi: 10.1016/j.jchromb.2006.12.003. Epub 2006 Dec 28.
A simple, sensitive and rapid liquid chromatography/tandem mass spectrometry (LC-MS/MS) method has been developed and validated to quantify griseofulvin in human plasma using propranolol hydrochloride as internal standard (IS). Samples were prepared using solid phase extraction and analysed without drying and reconstitution. The analytes were chromatographed on Hypersil, hypurity C18 reverse phase column under isocratic conditions using 0.05% formic acid in water:acetonitrile (30:70, v/v) as the mobile phase. Total chromatographic run time was 3.0 min. Quantitation was done on a triple quadrupole mass analyzer API-3000, equipped with turbo ion spray interface and operating in multiple reaction monitoring (MRM) mode to detect parent-->product ion transition for analyte and IS. The method was validated for sensitivity, matrix effect, accuracy and precision, linearity, recovery and stability studies. Linearity in plasma was observed over the concentration range 20-3000 ng/mL for griseofulvin. Lower limit of quantification (LLOQ) achieved was 20 ng/mL with precision (CV) less than 10% using 5 microL injection volume. The absolute recovery of analyte (87.36%) and IS (98.91%) from spiked plasma samples was consistent and reproducible. Inter-batch and intra-batch coefficients of variation across four validation runs (LLOQ, LQC, MQC and HQC) was less than 7.5%. The accuracy determined at these levels was within +/-4.2% in terms of relative error. The method was applied to a pilot bioequivalence study of 500 mg griseofulvin tablet in six healthy human subjects under fed condition.
已开发并验证了一种简单、灵敏且快速的液相色谱/串联质谱（LC-MS/MS）方法，以盐酸普萘洛尔作为内标（IS）定量测定人血浆中的灰黄霉素。使用固相萃取制备样品，无需干燥和复溶即可进行分析。分析物在Hypersil高纯C18反相柱上进行等度洗脱色谱分离，流动相为含0.05%甲酸的水：乙腈（30:70，v/v）。总色谱运行时间为3.0分钟。在配备涡轮离子喷雾接口并以多反应监测（MRM）模式运行的三重四极杆质谱分析仪API-3000上进行定量分析，以检测分析物和内标的母离子→子离子跃迁。该方法针对灵敏度、基质效应、准确度和精密度、线性、回收率和稳定性研究进行了验证。灰黄霉素在血浆中的线性范围为20-3000 ng/mL。使用5 μL进样体积时，定量下限（LLOQ）为20 ng/mL，精密度（CV）小于10%。加标血浆样品中分析物（87.36%）和内标（98.91%）的绝对回收率一致且可重复。四个验证批次（LLOQ、LQC、MQC和HQC）的批间和批内变异系数均小于7.5%。在这些水平下测定的准确度相对误差在±4.2%以内。该方法应用于500 mg灰黄霉素片在6名健康受试者进食条件下的初步生物等效性研究。
