A Sensitive and Specific Indirect Competitive Enzyme-Linked Immunosorbent Assay for Detection of Paeoniflorin and Its Application in Pharmacokinetic Interactions between Paeoniflorin and Glycyrrhizinic Acid

Planta Med. 2015 Jun;81(9):765-70. doi: 10.1055/s-0035-1545913. Epub 2015 Apr 9.

Abstract

This study aimed to develop an indirect competitive enzyme-linked immunosorbent assay based on monoclonal antibodies against paeoniflorin to study the effects of different doses of glycyrrhizinic acid on the pharmacokinetics of paeoniflorin in mice. An anti-paeoniflorin monoclonal antibody was produced from a hybridoma created through a fusion of splenocytes immunized with paeoniflorin-bovine serum albumin and conjugated with the hypoxanthine-aminopterin-thymidine-sensitive mouse myeloma cell line SP2/0. The resultant antibody was used to develop and validate a rapid, specific and sensitive, indirect competitive enzyme-linked immunosorbent assay for the measurement of paeoniflorin (linear range 4.8-312.5 ng · mL(-1)). The intraday and interday precision values of the indirect competitive ELISA method were well within the recommended range (≤ 10 %), and the recovery rate was, on average, 101.13 %. Pharmacokinetic parameters obtained from mouse blood samples at various intervals following the oral administration of paeoniflorin and glycyrrhizic acid at three doses (1 : 0.3, 1 : 1, 1 : 3, respectively) demonstrated that the highest dose of glycyrrhizic acid inhibits the absorption of paeoniflorin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Drug Interactions
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Female
  • Glucosides* / immunology
  • Glucosides* / isolation & purification
  • Glucosides* / pharmacokinetics
  • Glycyrrhizic Acid / pharmacokinetics*
  • Hybridomas / immunology
  • Mice
  • Mice, Inbred BALB C
  • Monoterpenes* / immunology
  • Monoterpenes* / isolation & purification
  • Monoterpenes* / pharmacokinetics

Substances

  • Antibodies, Monoclonal
  • Glucosides
  • Monoterpenes
  • peoniflorin
  • Glycyrrhizic Acid