Detection and determination of total amlodipine by high-performance thin-layer chromatography: a useful technique for pharmacokinetic studies

J Chromatogr B Biomed Appl. 1995 May 19;667(2):315-20. doi: 10.1016/0378-4347(95)00016-c.

Abstract

A novel analytical method for determination of the total plasma levels (free and protein bound) of the calcium channel blocking agent amlodipine has been developed using a high-performance thin-layer chromatographic (HPTLC) procedure. Detection and quantitation were performed without internal standards. In previously described methods for the estimation of amlodipine by gas chromatography and high-performance liquid chromatography, only the free levels in plasma and serum were quantified at 7% of the total amlodipine level, with the remaining 93% bound to plasma protein and tissue. The present method employs proteolysis of the plasma proteins by incubating plasma for 2 h in pepsin solution. After proteolysis amlodipine is extracted and a known amount of the extract is spotted on precoated silica-gel 60 F254 plates using a Camag Linomat IV autosampler. Amlodipine was quantified using a dual-wavelength TLC scanner. The method provides a direct estimate of the total amlodipine present in plasma.

MeSH terms

  • Amlodipine / administration & dosage
  • Amlodipine / blood*
  • Amlodipine / pharmacokinetics*
  • Chromatography, High Pressure Liquid / methods*
  • Humans
  • Pepsin A
  • Sensitivity and Specificity

Substances

  • Amlodipine
  • Pepsin A