Determination of glycine and threonine in topical dermatological preparations

J Pharm Biomed Anal. 2008 Aug 5;47(4-5):716-22. doi: 10.1016/j.jpba.2008.02.024. Epub 2008 Feb 29.

Abstract

In the present study, a single HPLC method was developed for the determination of glycine and threonine in cicatrizants. Two different preparations of a cream and an ointment, and the corresponding bandages, onto which the formulations were applied, were studied. The method involved matrix solubilisation with dichloromethane, liquid-liquid isolation of gly and thr with aqueous 1N NaOH, and derivatization with phenylisothiocyanate. Reversed-phase HPLC separation was carried out by gradient elution with 20mM aqueous NaClO4 and acetonitrile (from 90% to 30% aqueous NaClO4 in 10 min) on a LiChrospher 100 RP-18 cartridge (125 mm x 4.6 mm). Analytes were determined with a UV detector set at 245 nm. Quantitation was accomplished by internal standardization with methionine. Linearity was studied in the range 60-120% of the concentrations expected for gly and thr (viz. for gly from 200 to 400 microgml(-1), and for thr from 100 to 200 microgml(-1)). In reference aqueous samples, linear correlation (r) was better than 0.99 for gly and thr, while in spiked matrix samples r ranged from 0.97 to 0.98. Recoveries were in the 95-105% interval, and precision (CV%, N=6) was better than 5% for both analytes either in cream, ointment or bandages. The method was successfully used for the quality control of topical dermatological preparations.

MeSH terms

  • Acetonitriles / chemistry
  • Administration, Topical
  • Calibration
  • Chromatography, High Pressure Liquid / methods
  • Dermatologic Agents / analysis*
  • Excipients / chemistry
  • Glycine / analysis*
  • Isothiocyanates / chemistry
  • Methylene Chloride / chemistry
  • Ointments
  • Perchlorates / chemistry
  • Pharmaceutical Preparations / analysis*
  • Quality Control
  • Reference Standards
  • Sensitivity and Specificity
  • Sodium Compounds / chemistry
  • Spectrophotometry, Ultraviolet / methods
  • Threonine / analysis*
  • Water / chemistry

Substances

  • Acetonitriles
  • Dermatologic Agents
  • Excipients
  • Isothiocyanates
  • Ointments
  • Perchlorates
  • Pharmaceutical Preparations
  • Sodium Compounds
  • Water
  • phenylisothiocyanate
  • Threonine
  • Methylene Chloride
  • sodium perchlorate
  • Glycine
  • acetonitrile