Ben-Cha-Le-Ka-Wi-Chian remedy (BLW remedy) is a Thai traditional medicine that has long been used as an antipyretic drug by traditional practitioners and has been notified in the List of Medicine Products of the National List of Essential Drugs A.D. 2006. It is used as mixed powders of the roots of Capparis micracantha DC., Clerodendrum petasites S. Moore, Harrisonia perforata (Blanco) Merr., Ficus racemosa L. and Tiliacora triandra (Colebr.) Diels, in equal part by weight. From an exhaustive review and few reported data, the remedy has been contaminated and adulterated with upper ground of the plant used. Therefore, the quality of each root species, the pharmacognostic evaluation and multivariate analysis by 3D-HPLC were measured. The safety studies, cytotoxic acitivity, mutagenic testing and DNA damage using Brine shrimp method, Ames test and comet assay were investigated respectively, The efficacy study, antipyretic and analgesic activity by animal model, anti-mutagenic activity by Ames test, free radical scavenging activity by DPPH assay, cell proliferation by MTT assay and nitric oxide by Griess reagent assay were determined. Fourteen samples were collected from wild or non-cultivated places throughout Thailand. The main distinguishable features of five root species were obtained from the morphological and histological characters as well as TLC chromatogram. The histological results allowed an establishment of dichotomous key for the identification of each crude powdered species which is beneficial in resolving the adulteration and contamination of crude drugs in traditional medicine market. Three-dimensional of HPLC was showed clear twelve high major peaks in BLW remedy. All batches remedies were revealed a close relationship between batch 2 to 12 excepted batch 1. The Brine shrimp method demonstrated that most of samples are non-toxic except for the ethanol extract of T. triandra (LC50 44 ug/ml). Along with a no-direct mutagenic activity, however most of the extracts exhibited indirect mutagenic activity when combined with nitrosation. Nevertheless, the remedy extracts and the components herb extracts strongly inhibited mutagenicity when nitrite-treated I-aminopyrene was used as a mutagen. Only water and ethanol extract of C. micracantha and water extract of T. tnandra were exhibited higher damage in DNA as same as the positive control, H202. All doses of BLW remedy significantly (p<0.05) attenuated the increased rectal temperature produced by lipopolysaccharide (LPS) and were found to be as potent as acetylsalicylic acid (ASA). BLW remedy (400 mg/kg) also produced a significant analgesic response in the hot-plate test. Most of samples also showed good scavenging activity particularly in the ethanol extract samples. In case of cell proliferation, the entire samples were demonstrated LD% more than 2,000 ug/ml, whilst BLW remedy exhibited the LD50 more than 20,000 ug/ml. The scavenging activities on nitric oxide demonstrated that most of samples that prepared from each root species were demonstrated the optical density higher than vitamin C, while BLW remedy was exhibited lower optical density than vitamin C. Consequently, the present study provided further evidence to support the safety, efficacy and the quality of Thai traditional medicine: Ben-Cha-Lo-Ka-Wi-Chian remedy and its component herbs. Nevertheless, consumers should be advised on the adverse effects of using the remedy with nitrite containing foods. Moreover, the results of the current study could be described that not only each species which need to carry out for the safety, efficacy and quality, the combination of remedy were need to understand the consequences of such combined used also. Finally, this study helps clarifying the safety, efficacy and quality of each plant species and Ben-Cha-Lo-Ka-Wi-Chian remedy as well as providing additional scientific support for this well-known Thai traditional medicine.