NGHIÊN CỨU ỨNG DỤNG CÁC KỸ THUẬT SẮC KÝ TRONG PHÂN LẬP VÀ XÁC ĐỊNH ĐỘ TINH KHIẾT CỦA RUTIN TỪ NỤ HOA HÒE (Styphnolobium japonicum (L.) Schott)

Abstract

Background: Rutin is a flavonoid glycoside compound found in Hoa Hoe with a variety of important biological activities such as anti-oxidation, vascular protection, anti-cancer, cardiovascular protection and the demand for Styphnolobium japonicum buds in health care is more and more increasing. However, the current reference standard of rutin does not meet the market's demand in terms of supply, and price, especially purity. Based on the above reality, the research team decided to carry out the project "Research and application of chromatography techniques to isolate and determine the purity of rutin from Styphnolobium japonicum (L.) Schott buds". Objectives: To extract and purify rutin from buds and determine the purity of the isolated rutin using chromatography techniques. Material and methods: Rutin is extracted and crystallized at a suitable pH, then purified by dissolving with solvents of different polarity. The purified product was identified and determined using spectroscopic methods and modern techniques of UV-VIS, mass spectrometry, and HPLC-PDA. Results: From 50 grams of dried Styphnolobium japonicum (L.) Schott buds, 4.38 grams of high-purity rutin were crystallized and purified. Test results showed that X compound and the rutin standard had the same Rf (0.47) on thin-layer chromatography, with similar UV-VIS spectra at two specific absorption maxima λ_max 256 nm and 354 nm. Both the rutin standard and X compound produced mass spectra with a precursor ion of m/z = 609.15 Da and four characteristic fragment ions at m/z = 151; m/z = 255; m/z = 271; and m/z = 300.   The procedure for determining the purity of rutin using HPLC/PDA method was validated to meet the requirements according to ICH, with the purity assessment result reaching 95.66%. Conclusion: The study established a procedure for extracting and purifying rutin from Styphnolobium japonicum (L.) Schott buds using modern spectroscopic and chromatographic methods with high purity, without the need for column chromatography. This approach provides significant academic and economic benefits, contributing to the enrichment of reference substances and the enhancement of the quality of raw materials and products derived from buds of Hoa Hoe.