Validation of a high performance liquid chromatography method for the stabilization of epigallocatechin gallate

Int J Pharm. 2014 Nov 20;475(1-2):181-90. doi: 10.1016/j.ijpharm.2014.08.053. Epub 2014 Aug 28.

Abstract

Epigallocatechin gallate (EGCG) is a green tea catechin with potential health benefits, such as anti-oxidant, anti-carcinogenic and anti-inflammatory effects. In general, EGCG is highly susceptible to degradation, therefore presenting stability problems. The present paper was focused on the study of EGCG stability in HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) medium regarding the pH dependency, storage temperature and in the presence of ascorbic acid a reducing agent. The evaluation of EGCG in HEPES buffer has demonstrated that this molecule is not able of maintaining its physicochemical properties and potential beneficial effects, since it is partially or completely degraded, depending on the EGCG concentration. The storage temperature of EGCG most suitable to maintain its structure was shown to be the lower values (4 or -20 °C). The pH 3.5 was able to provide greater stability than pH 7.4. However, the presence of a reducing agent (i.e., ascorbic acid) was shown to provide greater protection against degradation of EGCG. A validation method based on RP-HPLC with UV-vis detection was carried out for two media: water and a biocompatible physiological medium composed of Transcutol®P, ethanol and ascorbic acid. The quantification of EGCG for purposes, using pure EGCG, requires a validated HPLC method which could be possible to apply in pharmacokinetic and pharmacodynamics studies.

Keywords: Catechins; Epigallocatechin gallate (EGCG); Reverse phase high performance liquid chromatography (RP-HPLC); Validation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Anti-Inflammatory Agents, Non-Steroidal / analysis*
  • Anti-Inflammatory Agents, Non-Steroidal / chemistry
  • Anticarcinogenic Agents / analysis*
  • Anticarcinogenic Agents / chemistry
  • Antioxidants / analysis*
  • Antioxidants / chemistry
  • Ascorbic Acid / chemistry
  • Buffers
  • Catechin / analogs & derivatives*
  • Catechin / analysis
  • Catechin / chemistry
  • Chemistry, Pharmaceutical / methods*
  • Chromatography, High Pressure Liquid
  • Chromatography, Reverse-Phase
  • Drug Stability
  • Drug Storage
  • Ethylene Glycols / chemistry
  • Excipients / chemistry*
  • HEPES / chemistry
  • Hot Temperature / adverse effects
  • Hydrogen-Ion Concentration
  • Oxidation-Reduction
  • Reducing Agents / chemistry
  • Solvents / chemistry
  • Spectrophotometry

Substances

  • Anti-Inflammatory Agents, Non-Steroidal
  • Anticarcinogenic Agents
  • Antioxidants
  • Buffers
  • Ethylene Glycols
  • Excipients
  • Reducing Agents
  • Solvents
  • Catechin
  • carbitol
  • epigallocatechin gallate
  • Ascorbic Acid
  • HEPES