Simple and fast LC-MS/MS method for quantification of terazosin in human plasma and application to bioequivalence study

J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Jan 15:1163:122517. doi: 10.1016/j.jchromb.2020.122517. Epub 2020 Dec 28.

Abstract

A simple, fast and sensitive LC-MS/MS method was developed to quantify terazosin in human plasma. The mobile phase consisted of acetonitrile-0.1% (v/v) formic acid (70:30, v/v). Prazosin was used as internal standard (IS). As deproteinization agent, acetonitrile produced a clean sample. A higher response intensity with more symmetrical peak was obtained using Agilent Poroshell 120 EC-C18 - Fast LC column (100 × 2.1mmID, 2.7 μm) compared with Kinetex XB-C18 (100 × 2.1 mm, 2.6 µm) column. The response of terazosin and IS were approximately two times in citrate phosphate dextrose (CPD) plasma compared with dipotassium ethylenediaminetetraacetic acid (K2EDTA) plasma. Plasma calibration curve was linear from 1.0 to 100.0 ng/mL, with coefficient of determination r2 ≥ 0.99. The within-run and between-run precision values (CV, %) were <5.2% and <7.8%, while accuracy values were 102.8-112.7% and 103.4-112.2%. The extended run accuracy was 98.6-102.8% and precision (CV, %) 4.3-10.4%. The recovery of analyte was >98% and IS >94%. Terazosin in plasma kept at benchtop was stable for 24 h, in autosampler tray for 48 h, in instrumentation room for 48 h, for 7 freeze-thaw cycles and in freezer for 140 days. Terazosin and IS stock standard solutions were stable for 140 days at room temperature and in the chiller. The high throughput method was successfully utilized to measure 935 samples in a bioequivalence study of terazosin.

Keywords: LC-MS/MS; Plasma; Prazosin; Protein precipitation; Terazosin.

Publication types

  • Randomized Controlled Trial

MeSH terms

  • Administration, Oral
  • Adolescent
  • Adult
  • Chromatography, High Pressure Liquid / methods*
  • Cross-Over Studies
  • High-Throughput Screening Assays
  • Humans
  • Linear Models
  • Male
  • Middle Aged
  • Prazosin / administration & dosage
  • Prazosin / analogs & derivatives*
  • Prazosin / blood
  • Prazosin / pharmacokinetics
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Tandem Mass Spectrometry / methods*
  • Therapeutic Equivalency
  • Young Adult

Substances

  • Terazosin
  • Prazosin