Sensitive and effective method with 96-well plate for determination of levamlodipine in human plasma using LC-MS/MS. 2024

Lingfang Dong, and Kun Li, and Shan Gao, and Wenyi Wang, and Yinghua Feng, and Jingying Zhang, and Jie Yang, and Weifeng Dong, and Wei Zhang
Department of Pharmacy, Henan Provincial People's Hospital, Zhengzhou University People's Hospital, Zhengzhou, Henan, 450003, China.

we developed an effective protein precipitation method for determination of levamlodipine in human plasma using LC-MS/MS. Sample extraction was carried out by using liquid-liquid extraction in 96-well plate format. (S)-Amlodipine-d4 was used as internal standard (IS). The chromatographic separation was achieved using Philomen Chiral MX (2) column (3 μm, 2.1×100 mm). Mobile phase A was comprised of Acetonitrile (ACN), Mono ethanol amine (MEA) and Iso-Propyl alcohol (IPA) (1000:1:10, v/v/v), Mobile phase B was IPA-ACN (2:1, v/v). The flow rate was 0.4 mL/min. The total run time of each sample was 4.0 min with gradient elution. LC-MS/MS spectra were generated in positive ion mode, and multiple reaction monitoring (MRM) was used to detect the following transitions: m/z 409.20 →238.15 for levamlodipine and 415.25→240.20 for (S)-Amlodipine-d4 (the IS). The method was linear from 50 -10000 pg/mL(R2=0.9988489),and the lower limit of quantification (LLOQ) was 50pg/mL. This method was applied to a bioequivalence study of levamlodipine.

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