An enzyme-linked immunosorbant assay using polyclonal antibodies against bacopaside I.
Bacopa monnieri (L.) Wettst. (Brahmi) is a medicinal plant used as a memory enhancer in Ayurvedic medicines. Its active components are triterpenoid glycosides namely pseudojujubogenin and jujubogenin glycosides. In order to analyze these saponin glycosides, an enzyme-linked immunosorbant assay (ELISA) was developed using polyclonal antibodies against bacopaside I, one of the pseudojujubogenin glycosides found in the plant. Bacopaside I was conjugated with a bovine albumin serum (BSA) to prepare an immunogen. The bacopaside I-BSA conjugate was immunized to a rabbit for producing polyclonal antibodies (PAbs). The results showed that the antibodies were raised specifically against pseudojujubogenin glycosides. An ELISA using anti-bacopaside I PAbs was performed in the range of 1.95-62.5 ng mL(-1) of bacopaside I and the limit of detection was 0.1 ng mL(-1). The method was validated and the applicability of the ELISA for analyzing saponin glycosides from Brahmi was demonstrated.
Phrompittayarat
Watoo
W
Department of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmaceutical Sciences, Naresuan University, Phitsanulok 65000, Thailand.
Putalun
Waraporn
W
Tanaka
Hiroyuki
H
Wittaya-Areekul
Sakchai
S
Jetiyanon
Kanchalee
K
Ingkaninan
Kornkanok
K
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006027:Glycosides / Q000302:isolation & purification; D018515:Plant Leaves / Q000737:chemistry; D018547:Plant Stems / Q000737:chemistry; D011817:Rabbits; D015203:Reproducibility of Results; D012503:Saponins / Q000276:immunology* / Q000302:isolation & purification*; D028083:Scrophulariaceae / Q000737:chemistry*; D012621:Seasons; D013785:Thailand; D014315:Triterpenes / Q000276:immunology* / Q000302:isolation & purification*
D000906:Antibodies; D006027:Glycosides; D012503:Saponins; D014315:Triterpenes; C476479:bacopaside IV; C476478:bacopaside V; C442170:bacopaside I; C442169:bacopaside II