Development of Anti-Asiaticoside Polyclonal Antibody-Based Immunoassay and Applications for Centella asiatica Products a
- PDF / 204,781 Bytes
- 8 Pages / 595.276 x 790.866 pts Page_size
- 62 Downloads / 194 Views
Development of Anti-Asiaticoside Polyclonal Antibody-Based Immunoassay and Applications for Centella asiatica Products and Human Serum Patcharin Tassanawat & Waraporn Putalun & Jukrapun Komaikul & Boonchoo Sritularak & Thaweesak Juengwatanatrakul & Hiroyuki Tanaka
Received: 21 November 2011 / Accepted: 13 February 2012 / Published online: 29 February 2012 # Springer Science+Business Media, LLC 2012
Abstract A sensitive indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed using polyclonal antibody (PAb) against asiaticoside (AS), one of the triterpenoid saponins found in Centella asiatica (Linn). ASbovine serum albumin conjugate was immunized to rabbits for producing PAb. The results showed that the antibodies were specific only for AS and very low specific for madecassoside whose basic skeleton is almost the same as AS. The range of the assay extends from 0.05–25 μg/ml of AS. A good correlation between ELISA and high-performance liquid chromatography methods was obtained when analysis of AS in the crude extracts of plant samples. In addition, the products containing C. asiatica in various preparations were determined AS content by competitive ELISA. The results showed that the product from tea bag preparation gave the highest yield of AS content (35.59 mg/g dry wt) comparing to other preparations. In order to evaluate the matrix effect of the serum for AS immunoassay, the standard curves of AS in different media were observed. Standard curve of the serum was similar to the water media and both curves P. Tassanawat : W. Putalun (*) : J. Komaikul Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand e-mail: [email protected] B. Sritularak Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand T. Juengwatanatrakul Faculty of Pharmaceutical Sciences, Ubon Ratchathani University, Ubon Ratchathani 34190, Thailand H. Tanaka Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-8582, Japan
showed the measurement range of 0.20–6.25 μg/ml. The developed ELISA method can be used for quality assessment of C. asiatica and their products including AS detection in serum samples. Keywords Asiaticoside . Centella asiatica . Enzyme-linked immunosorbent assay . Polyclonal antibody
Introduction Centella asiatica (Linn) (Umbelliferae) is a medicinal plant in Thailand, locally known as Bua-bok. Bua-bok juice is widely consumed in Thailand as a health drink for general well-being and as an herbal remedy for healing wounds. In addition, C. asiatica leaves are also used as vegetables in a variety of cuisines and traditional food recipes. The major chemical constituents from C. asiatica are triterpenoid saponins including asiaticoside (AS) and madecassoside (Fig. 1) (Inamdar et al. 1996). AS has been identified as the most active compound in the plant associated with the healing of wounds, neuroprotective, anti-inflammatory, antidepressant, protection of gastric ulcer, fungicidal, antibacterial and free radical scavenger (Cheng et al
Data Loading...
