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

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.

Novel immunoassay and rapid immunoaffinity chromatography method for the detection and selective extraction of naringin in Citrus aurantium

“…Purification or extraction of natural bioactive compounds based on IAC is a more concise, cheaper, environmentally friendly, and time‐saving strategy. Recently, we developed a potential method for the separation of glycyrrhizic acid and 20 ( R / S ) gisenoside epimers . In this study, the water extract of Citrus aurantium (CA‐EXT) was directly loaded onto an IAC after filtering, and then the column was washed with DW and eluted with 0.1 M glycine‐HCl buffer (pH 2.7) containing 0.5 M NaCl.…”

Study on the interaction of paeoniflorin with human serum albumin (HSA) by spectroscopic and molecular docking techniques

“…Therefore, the experimental results of the interaction between drugs and BSA cannot be completely identical with those of HSA. Although some spectroscopic studies on the interaction between paeoniflorin and bovine serum albumin (BSA) have been published [ 17 – 20 ], to our knowledge, a series of accurate and full basic data for clarifying the binding mechanisms of paeoniflorin to HSA remain unclear. Consequently, the binding characteristics of paeoniflorin with HSA including the quenching mechanism, quenching and binding constants were investigated in this study, by using fluorescence quenching method through the thermodynamic analysis.…”

Monoclonal Antibodies and Immunoassay for Medical Plant-Derived Natural Products: A Review

“…Zhao et al generated a new anti-paeoniflorin monoclonal antibody and developed a sensitive and specific ELISA to efficiently measure the concentration of paeoniflorin, and then applied it to explore the pharmacokinetics of paeoniflorin in the presence of GA at different doses [ 13 ].…”