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Journal of Thai Traditional & Alternative Medicine Vol. 7 No. 2 May-August (Supplement) 2009 ÒÒı

PP-6

Mutagenicity and Anti-mutagenicity of Ben-Cha-Lo-Ka-Wi-Chian Remedy

Using Ames Test

1 2 1,3
Chatubhong Singharachai , Oranuch Wongwattanasathien , Nijsiri Ruangrungsi

1
College of Public Health Sciences, Chulalongkorn University, Bangkok 10330, Thailand.
2
Faculty of Science and Technology, Rajabhat Maha Sarakham University, Maha Sarakam 44000, Thailand.
3
Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thai-
land.

Rational: According to the history of Thai traditional medicine use, Ben-Cha-Lo-Ka-Wi-Chian remedy,
that combines from five species of Tiliacora triandra (Colebr.) Diels, Capparis micracantha DC., Harrisonia

perforata (Blanco) Merr., Clerodendrum petasites S. Moore and Ficus racemosa L. roots, has also been used as
an antipyretic drug until present time. Although toxicity has never been reported after a long history use of Ben-
Cha-Lo-Ka-Wi-Chian remedy, research on the mutagenicity and anti-mutagenicity still needed to be investi-
gated. The presence of mutagen can be dangerous to human health due to mutational events involving in
several degenerative diseases such as cancer and arteriosclerosis.
Objective: To assess mutagenicity and anti-mutagenicity of Ben-Cha-Lo-Ka-Wi-Chian remedy and its
components, using Ames test.
Methodology: The roots of five species in Ben-Cha-Lo-Ka-Wi-Chian were collected from Nan province of
Thailand and authenticated by Ruangrungsi N. The voucher and number of specimens were deposited at
College of Public Health Sciences, Chulalongkorn University, Thailand. The air-dried and ground coarse powder
of five roots species were exhaustively extracted by maceration with absolute ethanol, then with water. The
ethanol extract was evaporated under vacuum and the water extract was lyophilized to dryness. The remedy
extract was prepared by mixing each extract in the quantity equivalent to those of the remedy preparation. The
ethanol extracts of each species and crude extract of the remedy were diluted in dimethylsulfoxide (DMSO) and
the water extracts were diluted in water at the adequate doses. Ames tests were performed on Salmonella
typhimurium strains TA98 for frame-shift mutation and TA100 for base-pair substitution mutation in an acidic
condition (pH 3 - 3.5) without metabolic activation. Both direct and in-direct mutagenicity were assayed in the

condition with and without sodium nitrite. The anti-mutagenicity against standard mutagen (nitrite treated 1-
aminopyrine) was also investigated.
Results: Ben-Cha-Lo-Ka-Wi-Chian remedy and its component extracts were non-mutagenic directly to-
wards S. typhimurium TA98 and TA100. On the contrary, most of the extracts showed indirect mutagenictiy
when combined with nitrosation. For anti-mutagenicity, most of the remedy extracts and the components
extracts exhibited strongly active inhibition (more than 60% inhibition) against nitrite treated 1-aminopyrine
induced mutagenicity in both TA98 and TA100. Whereas, the water extract of F.racemosa and C. petasites
were moderately active inhibitor (40 - 60 % inhibition).
Conclusion: The finding from the present study provided further evidence to support the safe consump-
tion of Thai traditional medicine: Ben-Cha-Lo-Ka-Wi-Chian remedy and its components. However, consumers
should beware of using the drug with nitrite containing food.