Simultaneous method development and validation of amlodipine besylate and hydrochlorothiazide in human plasma by lc-ms/ms
International Journal of Development Research
Simultaneous method development and validation of amlodipine besylate and hydrochlorothiazide in human plasma by lc-ms/ms
Received 26th July, 2017; Received in revised form 10th August, 2017 Accepted 17th September, 2017; Published online 10th October, 2017
Copyright ©2017, Meruva Sathish Kumar and Shanmugapandiyan. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
A rapid, simple, selective and sensitive LC-MS/MS method was developed for the determination of Amlodipine Besylate and Hydrochlorothiazide in human plasma using Amlodipine D4 and HCTZ 15N213C D2 as internal standard (IS). The method was developed with turbid ion spray (TIS) in the positive ion and multiple reaction monitoring (MRM) mode. The mobile phase was Water: Methanol (05:95) with 1 ml Ammonia Solution. Chromatographic separation was achieved on CHIRALCEL OZ-3R, 4.6 x 150 mm, column with a flow rate of 1.0ml/min. The MRM transitions monitored for Amlodipine Besylate and Hydrochlorothiazide and Amlodipine D4 and HCTZ 15N213C D2 were 296.00/205.00 (m/z), Mass 409.20/238.00 (m/z), Mass 301.00/207.00 (m/z), Mass 413.30/238.10(m/z) respectively. The developed method was validated as per FDA and ICH guidelines. Linearity was observed from of 51.910 pg/mL to 14464.765 pg/mL and0.509 ng/mL to 500789 ng/mL with correlation coefficient of 0.9969. The percent recovery for the drug and IS was found to be 45.5 and48.8 and,62.6 and 82.5% and respectively. Stability studies like freeze thaw, bench top, short term and long term were performed and the results were found to be within the acceptance limits according to FDA guidelines.
