Determination of iriflophenone 3-C-β-d-glucoside from Aquilaria spp. by an indirect competitive enzyme-linked immunosorbent assay using a specific polyclonal antibody.
Polyclonal antibody against iriflophenone 3-C-β-d-glucoside (IP3G), a major compound from the leaves of Aquilaria spp., was produced for the development of an enzyme-linked immunosorbent assay (ELISA). The results showed that the antibodies were specific for IP3G. The produced antibody has low cross reactivity with iriflophenone 3,5-C-β-d-diglucopyranoside (13%), genkwanin 5-O-β-primeveroside (3.55%) and no cross reactivity found in other compounds. The range of ELISA assay extends from 100 to 1560 ng/mL with coefficient of variation (CV) 1.19% to 2.07% for intra-assay and 3.76% to 7.15% for inter-assay precision levels. The recovery rates of IP3G in the leaves of Aquilaria spp. were in the range of 96.0% to 99.0% with CV 4.50% to 5.32%. A correlation between ELISA and high-performance liquid chromatography methods was obtained when analysis of IP3G in the plant samples (R(2) = 0.9321). These results suggest that the developed ELISA method can be applied to determine IP3G content with high specificity, rapidity, and simplicity. The developed immunosorbent assay in this study provides a useful tool for the analysis of IP3G in plant samples and products.
Faculty of Pharmaceutical Sciences, Khon Kaen Univ., Khon Kaen 40002, Thailand; Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), Natl. Research Univ.-Khon Kaen Univ., Khon Kaen 40002, Thailand.
Waraporn
W
Putalun
Gorawit
G
Yusakul
Paritad
P
Saensom
Boonchoo
B
Sritularak
Hiroyuki
H
Tanaka
D000906:Antibodies / Q000737:chemistry*; D001577:Benzophenones / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005419:Flavonoids / Q000032:analysis*; D005960:Glucosides / Q000032:analysis*; D010936:Plant Extracts / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D029645:Thymelaeaceae / Q000737:chemistry*
D000906:Antibodies; D001577:Benzophenones; D005419:Flavonoids; D005960:Glucosides; D010936:Plant Extracts
ELISA; analysis; chemical composition; diabetes