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

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry Pub Date : 2024-05-03 DOI:10.1016/j.ab.2024.115556

Lingfang Dong , Kun Li , Shan Gao , Wenyi Wang , Yinghua Feng , Jingying Zhang , Jie Yang , Weifeng Dong , Wei Zhang

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Abstract

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 to 10000 pg/mL（R²＝0.9988489）,and the lower limit of quantification (LLOQ) was 50 pg/mL. This method was applied to a bioequivalence study of levamlodipine.

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利用液相色谱-串联质谱/质谱（LC-MS/MS）技术，采用 96 孔板灵敏有效地测定人血浆中的左旋氨氯地平

我们建立了一种有效的蛋白质沉淀法，利用 LC-MS/MS 测定人血浆中的左旋氨氯地平。样品提取采用96孔板液液萃取法。(S)-Amlodipine-d4 用作内标（IS）。色谱分离采用 Philomen Chiral MX (2) 色谱柱（3 μm，2.1 × 100 mm）。流动相 A 由乙腈 (ACN)、单乙醇胺 (MEA) 和异丙醇 (IPA) 组成（1000:1:10，v/v/v），流动相 B 为 IPA-ACN（2:1，v/v）。流速为 0.4 mL/min。每个样品的梯度洗脱总运行时间为 4.0 分钟。LC-MS/MS 图谱在正离子模式下生成，多反应监测（MRM）用于检测以下跃迁：左旋氨氯地平的跃迁为 m/z 409.20 → 238.15，(S)-阿莫地平-d4（IS）的跃迁为 m/z 415.25 → 240.20。该方法在50～10000 pg/mL范围内线性关系良好（R2＝0.9988489），定量下限为50 pg/mL。该方法应用于左旋氨氯地平的生物等效性研究。

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来源期刊

Analytical biochemistry 生物-分析化学

CiteScore

5.70

自引率

0.00%

发文量

283

审稿时长

44 days

期刊介绍： The journal''s title Analytical Biochemistry: Methods in the Biological Sciences declares its broad scope: methods for the basic biological sciences that include biochemistry, molecular genetics, cell biology, proteomics, immunology, bioinformatics and wherever the frontiers of research take the field. The emphasis is on methods from the strictly analytical to the more preparative that would include novel approaches to protein purification as well as improvements in cell and organ culture. The actual techniques are equally inclusive ranging from aptamers to zymology. The journal has been particularly active in: -Analytical techniques for biological molecules- Aptamer selection and utilization- Biosensors- Chromatography- Cloning, sequencing and mutagenesis- Electrochemical methods- Electrophoresis- Enzyme characterization methods- Immunological approaches- Mass spectrometry of proteins and nucleic acids- Metabolomics- Nano level techniques- Optical spectroscopy in all its forms. The journal is reluctant to include most drug and strictly clinical studies as there are more suitable publication platforms for these types of papers.

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