Simultaneous determination of atorvastatin, amlodipine, ramipril and benazepril in human plasma by LC-MS/MS and its application to a human pharmacokinetic study.
A rapid, simple, sensitive and specific LC-MS/MS method has been developed and validated for the simultaneous estimation of atorvastatin (ATO), amlodipine (AML), ramipril (RAM) and benazepril (BEN) using nevirapine as an internal standard (IS). The API-4000 LC-MS/MS was operated under the multiple-reaction monitoring mode using electrospray ionization. Analytes and IS were extracted from plasma by simple liquid-liquid extraction technique using ethyl acetate. The reconstituted samples were chromatographed on C(18) column by pumping 0.1% formic acid-acetonitrile (15:85, v/v) at a flow rate of 1 mL/min. A detailed validation of the method was performed as per the FDA guidelines and the standard curves were found to be linear in the range of 0.26-210 ng/mL for ATO; 0.05-20.5 ng/mL for AML; 0.25-208 ng/mL for RAM and 0.74-607 ng/mL for BEN with mean correlation coefficient of ≥0.99 for each analyte. The intra-day and inter-day precision and accuracy results were well with in the acceptable limits. A run time of 2.5 min for each sample made it possible to analyze more than 400 human plasma samples per day. The developed assay method was successfully applied to a pharmacokinetic study in human male volunteers.
Pilli
Nageswara Rao
NR
University College of Pharmaceutical Sciences, Jawaharlal Nehru Technological University, Kukatpally, Hyderabad, India.
Inamadugu
Jaswanth Kumar
JK
Mullangi
Ramesh
R
Karra
Vijaya Kumari
VK
Vaidya
Jayathirtha Rao
JR
Rao
J V L N Seshagiri
JV
D017311:Amlodipine / Q000097:blood* / Q000493:pharmacokinetics; D000924:Anticholesteremic Agents / Q000097:blood / Q000493:pharmacokinetics; D000959:Antihypertensive Agents / Q000097:blood / Q000493:pharmacokinetics; D000069059:Atorvastatin; D001552:Benzazepines / Q000097:blood* / Q000493:pharmacokinetics; D002853:Chromatography, Liquid; D004355:Drug Stability; D006538:Heptanoic Acids / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D016018:Least-Squares Analysis; D008297:Male; D019829:Nevirapine / Q000032:analysis; D011758:Pyrroles / Q000097:blood* / Q000493:pharmacokinetics; D017257:Ramipril / Q000097:blood* / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*
D000924:Anticholesteremic Agents; D000959:Antihypertensive Agents; D001552:Benzazepines; D006538:Heptanoic Acids; D011758:Pyrroles; D017311:Amlodipine; D019829:Nevirapine; D000069059:Atorvastatin; D017257:Ramipril; C044946:benazepril