An Enzyme-linked Immunosorbent Assay for Genistein 7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside Determination in Derris scandens using a Polyclonal Antibody

Kamonthip Jutathis, Tharita Kitisripanya, Orain Udomsin, Chadathorn Inyai, Boonchoo Sritularak, Hiroyuki Tanaka, Waraporn Putalun

Research output: Contribution to journalArticle

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Abstract

Introduction: Genistein 7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside (GTG) is a major bioactive compound in Derris scandens. It is responsible for anti-inflammatory activity by inhibition of cyclooxygenase and lipoxygenase. There are many commercial products of D. scandens available in Thailand. Objective: To develop an enzyme-linked immunosorbent assay (ELISA) for the quantitative analysis of GTG in plant material and derived products using a polyclonal antibody. Methods: An immunogen was synthesised by conjugating GTG with a carrier protein. The polyclonal antibody against GTG (GTG-PAb) was produced in New Zealand white rabbits. The ELISA method was validated for specificity, sensitivity, accuracy, precision and correlation with HPLC. Results: The polyclonal antibody was specific to GTG and genistin within the range of compounds tested. The GTG ELISA was applied in the range 0.04–10.00 μg/mL with a limit of detection of 0.03 μg/mL. The recovery of GTG in spiked Derris scandens extracts ranged from 100.7 to 107.0%, with a coefficient of variation less than 7.0%. The intra- and inter-assay variations were less than 5.0%. The ELISA showed a good correlation with HPLC-UV analysis for GTG determination in samples, with a coefficient of determination (r2) of 0.9880. Conclusion: An ELISA was established for GTG determination in Derris scandens. The GTG-PAb can react with GTG and genistin, but genistin has not been found in the plant. Therefore, the ELISA can be used for high throughput quality control of GTG content in D. scandens and its products.

Original languageEnglish
Pages (from-to)336-342
Number of pages7
JournalPhytochemical Analysis
Volume27
Issue number6
DOIs
Publication statusPublished - Nov 1 2016

Fingerprint

Derris
Immunosorbents
Genistein
genistein
polyclonal antibodies
Assays
Enzyme-Linked Immunosorbent Assay
enzyme-linked immunosorbent assay
genistin
Antibodies
Enzymes
High Pressure Liquid Chromatography
Lipoxygenase
New Zealand White rabbit
transport proteins
assays
prostaglandin synthase
Thailand
Prostaglandin-Endoperoxide Synthases
lipoxygenase

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Food Science
  • Biochemistry
  • Molecular Medicine
  • Plant Science
  • Drug Discovery
  • Complementary and alternative medicine

Cite this

An Enzyme-linked Immunosorbent Assay for Genistein 7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside Determination in Derris scandens using a Polyclonal Antibody. / Jutathis, Kamonthip; Kitisripanya, Tharita; Udomsin, Orain; Inyai, Chadathorn; Sritularak, Boonchoo; Tanaka, Hiroyuki; Putalun, Waraporn.

In: Phytochemical Analysis, Vol. 27, No. 6, 01.11.2016, p. 336-342.

Research output: Contribution to journalArticle

Jutathis, Kamonthip ; Kitisripanya, Tharita ; Udomsin, Orain ; Inyai, Chadathorn ; Sritularak, Boonchoo ; Tanaka, Hiroyuki ; Putalun, Waraporn. / An Enzyme-linked Immunosorbent Assay for Genistein 7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside Determination in Derris scandens using a Polyclonal Antibody. In: Phytochemical Analysis. 2016 ; Vol. 27, No. 6. pp. 336-342.
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abstract = "Introduction: Genistein 7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside (GTG) is a major bioactive compound in Derris scandens. It is responsible for anti-inflammatory activity by inhibition of cyclooxygenase and lipoxygenase. There are many commercial products of D. scandens available in Thailand. Objective: To develop an enzyme-linked immunosorbent assay (ELISA) for the quantitative analysis of GTG in plant material and derived products using a polyclonal antibody. Methods: An immunogen was synthesised by conjugating GTG with a carrier protein. The polyclonal antibody against GTG (GTG-PAb) was produced in New Zealand white rabbits. The ELISA method was validated for specificity, sensitivity, accuracy, precision and correlation with HPLC. Results: The polyclonal antibody was specific to GTG and genistin within the range of compounds tested. The GTG ELISA was applied in the range 0.04–10.00 μg/mL with a limit of detection of 0.03 μg/mL. The recovery of GTG in spiked Derris scandens extracts ranged from 100.7 to 107.0{\%}, with a coefficient of variation less than 7.0{\%}. The intra- and inter-assay variations were less than 5.0{\%}. The ELISA showed a good correlation with HPLC-UV analysis for GTG determination in samples, with a coefficient of determination (r2) of 0.9880. Conclusion: An ELISA was established for GTG determination in Derris scandens. The GTG-PAb can react with GTG and genistin, but genistin has not been found in the plant. Therefore, the ELISA can be used for high throughput quality control of GTG content in D. scandens and its products.",
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AU - Udomsin, Orain

AU - Inyai, Chadathorn

AU - Sritularak, Boonchoo

AU - Tanaka, Hiroyuki

AU - Putalun, Waraporn

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