Bülow, F.E., et al. A novel thin layer chromatography based method for the quantification of quillaic acid saponins. J Pharm Biomed Anal, 2026 Jun 15, 273:117409. PMID: 41687224

This paper presents a novel, cost-effective thin-layer chromatography (TLC) -based method for quantifying quillaic acid saponins, key bioactive compounds with significant medicinal and immunological applications, such as vaccine adjuvants and endosomal escape enhancers. Traditional analytical techniques like HPLC have high costs and resource requirements. By introducing fluorescence tagging with NBD-H and validating the method against established HPLC techniques, the study offers a practical, accessible tool that could enhance quality control, plant cultivation monitoring, and research on saponin-based compounds worldwide.

The technique involves a simple TLC method. The process begins with acidic hydrolysis of saponins to release the sapogenin quillaic acid, which is then derivatized using the fluorescent reagent NBD-H to form hydrazones. These fluorescent compounds are separated on silica gel TLC plates using a specific solvent system. Detection is performed at a wavelength of 480 nm with densitometry, allowing for quantification of quillaic acid based on the intensity of the fluorescent bands. The method is validated with mass spectrometry confirmation and validated parameters such as linearity and sensitivity.

Comment from DKI: This study presents an open access, cost-effective TLC-based method that offers an inexpensive and accessible tool for the quantification of quillaic acid saponins, enabling laboratories with limited resources to perform phytochemical analysis and quality control.

Link: https://pubmed.ncbi.nlm.nih.gov/41687224/