Method Development and Validation of
Liquid Chromatography-Tandem Mass Spectrometry for Angiotensin-II in Human Plasma:
Application to Study Interaction Between Atorvastatin & Olmesartan Drug
Combination |
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| Authors: | Rakesh Das Tapan Kumar Pal |
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| Institution: | .Department of Pharmaceutical Technology, Jadavpur University, Kolkata, WB India ;.Bioequivalence Study Center, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700032 WB India |
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| Abstract: | Simple and sensitive Liquid Chromatography-Tandem Mass Spectrometry (LCMS/MS)
method was developed and validated, then was implicated on hypertensive human
subjects to study drug interaction of atorvastatin (ATVS) and Olmesartan (OLM) on
status of Angiotensin-II (ANG-II). The ANG-II in plasma was extracted with 5 mL
methanol containing 5 % formic acid through C18 (cartridges)
liquid–liquid extraction, dried and reconstituted with 1 mL of 16 % acetonitrile in
0.1 % formic acid in water. The chromatographic separation of ANG-II with a Agilent
technology 6410 Triple quadrupole was carried multiple reaction monitoring scan mode
with a Agilent 1290 Infinity LC system for UHPLC. The sample were separated on a
(Thermo Scientific) Hy-Purity advance (50 × 4.6 mm, 5 μm) using Mobile Phase A: 16 %
acetonitrile in 0.1 % formic acid in water and Mobile Phase B: 0.1 % formic acid in
methanol at a flow rate of 0.3 mL/min, performed at ambient temperature. The mobile
phase gradient of 16 % acetonitrile in water was linearly increased to 38 %
acetonitrile over 10 min and subsequently the mobile-phase was increased to 100 %
acetonitrile over 15 min. The developed method was validated for specificity,
accuracy, precision, stability, linearity, sensitivity and recovery. The method was
linear between peak area ratio of standard and internal standard over the range of
50–800 ng/mL. The method was successfully applied for the drug interaction study
revealed levels of ANG-II were significantly higher in ATVS + OLM treatment
condition as compared to individual treatment of OLM. This reflects the reason of
low effectiveness of ATVS + OLM in combination instead of synergistic
activity.Electronic supplementary materialThe online version of this article (doi:10.1007/s12291-014-0457-x) contains supplementary material, which is available to authorized
users. |
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| Keywords: | Angiotensin-II Angiotensin-III LCMS/MS Drug interaction Olmesartan Atorvastatin |
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