Determination of Diosgenin Content in Medicinal Plants with Enzyme-Linked Immunosorbent Assay

Abstract

Many medicinal plants contain diosgenin, which has a significant medicinal value. However, there is currently no effective and rapid analytical method to determine the diosgenin content of plants or products. In the present work we have developed an indirect competitive enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of diosgenin in herbal medicines. Diosgenin was conjugated with bovine serum albumin (BSA) for immunization. A polyclonal antibody developed in rabbits against a diosgenin-BSA conjugate was shown to be specific for diosgenin. The developed ELISA assay was highly sensitive, specific, and easy to perform. In addition, it gave more precise results with less variation than other methods that have been used in the past, including gravimetric and spectrophotometric assays, and correlated well with high-performance liquid chromatography. The diosgenin content determined by ELISA varied widely, with the highest and lowest values in rhizomes or tubers of Paris polyphylla and Dioscorea opposita Thunb. "Jiao-ban Yam", respectively, differing by more than 9000-fold. These results suggest that the ELISA method can be used as a rapid, simple, sensitive, and accurate tool for quantitative analysis of samples containing diosgenin, and may provide an important criterion for quality evaluation and a valuable tool for quality control of diosgenin-containing medicinal plants.

Diosgenin: Recent Highlights on Pharmacology and Analytical Methodology

“…In this field, Li et al [129] developed an indirect competitive ELISA method to quantify diosgenin, namely, in Paris and Dioscorea species. The diosgenin molecule, which is too small to be considered an immunogen, was conjugated with bovine serum albumin (BSA) to create the immunization.…”

“…Then, from rabbits, a specific polyclonal antibody was developed against diosgenin-BSA conjugate. This method allowed a screening of several Chinese plants which have diosgenin as component [129]. A similar strategy was also developed to detect and quantify sarsasapogenin, a steroidal sapogenin structurally similar to diosgenin.…”

Application of High‐speed Countercurrent Chromatography–Evaporative Light Scattering Detection for the Separation of Seven Steroidal Saponins from Dioscorea villosa

“…A variety of chemical constituents in Dioscorea species have been reported, comprising diterpenoids, flavonoids, phenanthrenes, sterols, steroidal saponins and stilbenes (Yang et al , ). Among these constituents, steroidal saponins, representative secondary metabolites of Dioscorea species, are being used to control the quality of botanical products or dietary supplements containing Disocorea species (Liu et al , ; Li et al , ). Thus, there seems to be a great need for an efficient separation method for steroidal saponins of Dioscorea species as these compounds may be utilised for pharmacological probes and quality control of botanical preparations (Li et al , ; Lin et al , ; Liu et al , ; Lin and Yang, ).…”

Diosgenin From Dioscorea Nipponica Rhizoma Against Graves’ Disease—On Network Pharmacology and Experimental Evaluation

“…The content of diosgenin in Medicinal Plants could be determinated accurately by enzyme-linked immunosorbent assay or a micellar electrokinetic capillary chromatographic (MECC) (J. Li et al, 2010 ; N. Wang et al, 2019 ), and the water contents in diosgenin could be assayed with high accuracy using the mathematical model of near-infrared spectroscopy technology (M. Zhang et al, 2019 ). For the past few years, the biological activities of diosgenin has drawn much attention by studied in vivo and in vitro ( Leger et al, 2004 ; J.; Liagre et al, 2004 ; M. J.; Liu et al, 2005 ; Liagre et al, 2004 ; Srinivasan et al, 2009 ; Li et al, 2010 ; Ru et al, 2014 ; Chao et al, 2017 ; Qiao et al, 2018 ; Kessler et al, 2019 ; N.; Wang et al, 2019 ; M. Zhang et al, 2019 ; Mohamadi-Zarch et al, 2020 ).…”

“… Li et al, 2010 ; N. Wang et al, 2019 ), and the water contents in diosgenin could be assayed with high accuracy using the mathematical model of near-infrared spectroscopy technology (M. Zhang et al, 2019 ). For the past few years, the biological activities of diosgenin has drawn much attention by studied in vivo and in vitro ( Leger et al, 2004 ; J.; Liagre et al, 2004 ; M. J.; Liu et al, 2005 ; Liagre et al, 2004 ; Srinivasan et al, 2009 ; Li et al, 2010 ; Ru et al, 2014 ; Chao et al, 2017 ; Qiao et al, 2018 ; Kessler et al, 2019 ; N.; Wang et al, 2019 ; M. Zhang et al, 2019 ; Mohamadi-Zarch et al, 2020 ). In previous studies, we found that diosgenin ( Mu et al, 2012 ) promoted apoptosis of human primary thyroid cells under the action of IGF-1 through the caspase signaling pathway, but its specific mechanism is not clear.…”