Determination of (-)-epigallocatechin-3-gallate octaacetate and its metabolites in plasma of rats for pharmacokinetic study by ultra-performance-liquid-chromatography coupled to quadrupole-time-of-flight-mass-spectrometry

Kai On Chu; Gene Chi Wai Man; Sze Wan Hung; Tak Hang Chan; Wai Yip Thomas Lee; Kwok Ping Chan; Chi Pui Pang; Chi Chiu Wang

doi:10.3389/fphar.2022.1025053

Determination of (-)-epigallocatechin-3-gallate octaacetate and its metabolites in plasma of rats for pharmacokinetic study by ultra-performance-liquid-chromatography coupled to quadrupole-time-of-flight-mass-spectrometry

Front Pharmacol. 2022 Oct 11:13:1025053. doi: 10.3389/fphar.2022.1025053. eCollection 2022.

Authors

Kai On Chu^{1

2}, Gene Chi Wai Man², Sze Wan Hung², Tak Hang Chan^{3

4}, Wai Yip Thomas Lee⁵, Kwok Ping Chan¹, Chi Pui Pang^{1

6}, Chi Chiu Wang^{2

7}

Affiliations

¹ Department of Ophthalmology and Visual Sciences, The Chinese University of Hong Kong, Shatin, China.
² Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, Shatin, China.
³ Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong, China.
⁴ Department of Chemistry, McGill University, Montreal, QC, Canada.
⁵ Aptorum Group Limited, Hong Kong, China.
⁶ Joint Shantou International Eye Center of Shantou University and the Chinese University of Hong Kong, Shantou, China.
⁷ Li Ka Shing Institute of Health Sciences; and School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, China.

Abstract

(-)-Epigallocatechin-gallate octaacetate (pro-EGCG), a prodrug of epigallocatechin-gallate (EGCG), has been used for pre-clinical study for the treatment of endometriosis. A validated analytical method has been developed for the determination of plasma pro-EGCG and its metabolites after oral administration using ultra-performance-liquid-chromatography coupled to quadrupole-time-of-flight-mass-spectrometry (UPLC-Qtof-MS). This method is more robust, rapid, sensitive, simpler, and able to detect pro-EGCG metabolites compared to our previous method. Pro-EGCG in the plasma was stabilized from rapid degradation by formic acid, extracted by isopropanol/methyl-tert-butyl ether mixture, separated by UPLC core column, and quantified by an exact mass method with Qtof-MS. The lower limit of quantification (LLOQ), intra-day and inter-day precision, and accuracy for the range of 0.01-2.5 μg/mL were within acceptable limits. The sensitivity was improved by 25 folds using pro-EGCG ammonium adduct [M + NH4]⁺. This is the first report on the pharmacokinetics of oral administration with maximum-concentration (Cmax) was 0.067 ± 0.04 μg/mL, time-of-maximum-concentration (Tmax) was 1.33 h, area-under-curve (AUC) was 0.20 ± 0.05 h × µg/mL, and elimination-rate was 0.20 ± 0.11 hr^-1. The pharmacokinetic profiles of pro-EGCG metabolites, (-)-epigallocatechin-gallate (EGCG) diacetates and EGCG triacetates, were also presented. This method is robust, rapid, and sensitive for the pharmacokinetic study of pro-EGCG and metabolites.

Keywords: (-)-Epigallocatechin-3-gallate-octaacetate; LC-MS; determination; pharmacokinetics; validation.