QUANTITATION OF AMLODIPINE IN HUMAN PLASMA BY LCMS/MS ASSAY
Keywords:
Amlodipine, Tizanidine, Human plasma, LC-MSMSAbstract
ÂÂÂÂÂÂÂÂÂÂÂÂÂÂÂÂÂObjective: To develop and validate a simple, precise, and rapid liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for quantification of amlodipine in human plasma.
Methods: Chromatographic analysis was performed on Atlantis dC18 column (2.1 x 100 mm, 3 µm) with a mobile phase consisting of acetonitrile and 10 mM formic acid (80:20, v: v) that was delivered at a flow rate of 0.3 ml/min. The eluents were monitored using electrospray ionization in the positive ion mode set at transition 409 → 238.4 and 254.3 → 43.9 for amlodipine and tizanidine hydrochloride (IS), respectively. The method was validated for linearity, accuracy, precision, and recovery as per US-FDA guidelines.
Results: The retention times of amlodipine and tizanidine (IS) were 1.26 and 1.22 respectively. The relationship between amlodipine concentration and peak height ratio of amlodipine to the IS was linear (R2³ 0.9868) in the range of 0.2–20 ng/ml, and the intra-and inter-day coefficient of variations and bias were ≤14.4% and ≤13.6% and ≤13.7% and ≤11.2%, respectively.
Conclusion: The proposed method is simple, precise, and accurate for rapid measurement of amlodipine level using 0.5 ml human plasma. Further, the assay was successfully applied to determine amlodipine level in human plasma samples obtained from a healthy volunteer.
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References
Haria M, Wagstaff AJ. Amlodipine a reappraisal of its pharmacological properties and therapeutic use in cardiovascular disease. Drugs 1995;50:560-86.
Abernethy DR. The pharmacokinetic profile of amlodipine. Am Heart J 1989;118:1100-3.
Meredith PA, Elliott HL. Clinical pharmacokinetics of amlodipine. Clin Pharmacokinet 1992;22:22-31.
Ramadan NK, Mohamed HM, Moustafa AA. Rapid and highly sensitive HPLC and TLC methods for quantification of amlodipine besilate and valsartan in bulk powder and in pharmaceutical dosage forms and in human plasma. Anal Lett 2010;43:570-81.
Krol GJ, Noe AJ, Yeh SC, Raemsch KD. Gas and liquid chromatographic analyses of nimodipine calcium antagonist in blood plasma and cerebrospinal fluid. J Chromatogr 1984;305:105-18.
Bereford AP, Marae PV, Stopher DA, Wood BA. Analysis of amlodipine in human plasma by gas chromatography. J Chromatogr Biomed Appl 1987;420:178-83.
Pandya KK, Saita M, Gandhi TP, Modi IA, Modi RI, Chakravarthy BK. Detection and determination of total amlodipine by high-performance thin-layer chromatography. J Chromatogr B: Biomed Sci Appl 1995;667:315-20.
Zarghi A, Foroutan SM, Shafaati A, Khoddam A. Validated HPLC method for determination of amlodipine in human plasma and its application to pharmacokinetic studies. IL Farmaco 2005;60:789-92.
Meyyanathan SN, Muralidharan S, Rajan S, Gopal K, Suresh B. A simple preparation with HPLC-UV method for estimation of amlodipine from plasma: application to bioequivalence study. Open Chem Biomed Methods J 2008;1:22-7.
Bahrami Gh, Mirzaeei Sh. Simple and rapid method for determination of amlodipine in human serum with fluorescence detection and its use in pharmacokinetic studies. J Pharm Biomed Anal 2004;36:163-8.
Sevgi T, Sedef A. Determination of amlodipine in human plasma by high performance liquid chromatography with fluorescence detection. J Chromatogr B 2001;758:305-10.
Josefsson M, Zackrisson AL, Norlander B. Sensitive high-performance liquid chromatographic analysis of amlodipine in human plasma with amperometric detection and a single-step solid-phase sample preparation. J Chromatogr B: Biomed Sci Appl 1995;672:310-3.
Prasad Bathula SNV, Prasad KD. Quantitative determination of amlodipine in human plasma by ultra-performance liquid chromatography-electrospray ionization mass spectrometry: application to a clinical pharmacokinetic study. Asain J Pharm Clin Res 2012;5:89-93.
Massaroti P, Moraes LA, Marchioretto MA, Cassiano NM, Bernasconi G, Calafatti SA, et al. Pedrazzoli. J Anal Bioanal Chem 2005;382:1049-54.
Wang X, Chaunmin WEI, Chanmei LV, Wang B, Ruichen GUO. An accurate, rapid and sensitive LC-MS-MS method for quantification of amlodipine in human plasma: application to a bioequivalence study. Latt Am J Pharm 2013;32:1355-66.
Dams R, Huestis MA, Lambert WE, Murphy CM. Matrix effect in bio-analysis of illicit drugs with LC-MS-MS: influence of ionization type, sample preparation, and biofluid. J Am Soc Mass Spectrom 2003;14:1290-4.
Guidance for Industry Bioanalytical Method Validation†US Department of Health Services, Food and Drug Administration, CDER, CVM; 2001.