QUALITY SAFETY AND EFFICACY EVALUATION OF CHA TU KA LA THAD AND TREE PHON THAD REMEDIES

Abstract

Cha Tu Ka La Thad (CKT) and Tree Phon Thad (TPT) remedies are the remedies of traditional Thai medicine recorded in Tamra Paetsat Songkhrau. CKT remedy is composed of four species (i.e., Plumbago indica, Acorus calamus, Clerodendrum paniculatum and Dolichandrone serrulata) and each in contains equal part by weight. TPT remedy consists of three species (i.e., Zingiber zerumbet, Zingiber montanum and Cymbopogon nardus) and each contains in equal part by weight. Both remedies have been used as antipyretic and anti-inflammatory indications for a long time. This study was carried out to provide pharmacognostic specification of each species in CKT and TPT remedies. In addition, the ethanol and fractionated water extracts of both remedies were investigated for the safety and efficacy in vitro and in vivo. The macroscopic and microscopic, physico-chemical parameters, and chemical fingerprints were performed to estabish the standard parameters of each species in CKT and TPT remedies. The mutagenic and antimutagenic activities of all extracts were studied using Ames test with pre-incubation method. Most extracts without nitrite treatment exhibited non-mutagenicity toward Salmonella typhimurium TA98 and TA100, except the ethanol extract of A. calamus which exhibited mutagenicity to both strains and the fractionated water extracts of P. indica and C. nardus showed slightly mutagenicity toward S. typhimurium TA98. However, most of the extracts were mutagenic on both strains of S. typhimurium after being treated with sodium nitrite. Additionally, most of the extracts exhibited strong antimutagenic potential against the nitrite-treated 1- aminopyrene under acidic condition in the Ames test. Among CKT and TPT remedies and their ingredient extracts, results demonstrated the ethanol extract of A. calamus indicated the highest toxicity against brine shrimp with LC50 of 129.88 µg/ml. The fractionated water extracts of A. calamus and P. indica showed the highest DNA damage in human lymphocytes using comet test with total score of 233.67 and 158.33 respectively, while CKT and TPT remedies presented non-genotoxicity in comet test. These studies demonstrated the ethanol extracts of Z. montanum, A. calamus and the fractionated water extracts of P. indica, CKT and TPT remedies showed the highest antioxidant capacity in various in vitro models. Furthermore, all doses of CKT and TPT remedy extracts demonstrated anti-inflammatory activities.