New application of sepaflash ® reversed phase C18 purification column -- purification and preparation of anthocyanins

Anthocyanidin, or anthocyanin, of Xubo applied technology research center of Qiu Wenjun, has a chemical formula of c15h11o6 (its basic structural formula is shown in Figure 1) It is a water-soluble plant pigment, which exists in the cell fluid inside the vacuole It is related to the color of flowers and the redness of leaves Anthocyanins are widely found in the cores, skins or seeds of various plants, such as grape, pine bark, apricot shell, cherry blossom, hawthorn leaf, spruce leaf, etc At present, most of the pigments used in the food industry are synthetic pigments, almost all of which have different degrees of toxicity Long-term consumption will harm people's health Therefore, natural pigments have gradually attracted the attention of researchers As a natural food pigment, anthocyanin is not only safe, non-toxic and rich in resources, but also has certain nutritional and pharmacological effects The results show that proanthocyanidins have the functions of antioxidant, free radical scavenging, liver protection and detoxification, blood pressure reduction, blood lipid reduction, anti-inflammatory, anti-cancer, cardiovascular disease treatment, etc., and have great application potential in food, cosmetics, medicine, etc Fig 1 Chemical structure of anthocyanin Mr Zou from the school of food, Changzhou University is currently working on extracting anthocyanin from plant testa or leaves In the early stage, traditional ethanol extraction method was used After preliminary extraction, it was found that the extract contains not only main components but also a small amount of water-soluble impurities The HPLC analysis map is shown in Fig 2 In order to remove these impurities and further improve the purity of the extracted anthocyanin, Mr Zou contacted the application technology research center of Santai technology, hoping that we can help him solve this problem Fig 2 After understanding the demand and sample properties of the other party, Qiu Wenjun, the application engineer, decided to use sepaflash ® series C18 reversed-phase purification column for purification and preparation The sample is liquid ethanol extract of plant seed coat After simple centrifugation, take 5ml of supernatant for sample injection The experimental parameters of flash preparation are shown in Table 1 Table 1 Sepabean ® machine chromatographic column 40g sepaflash ® bonded series C18 reversed-phase purification column (spherical silica gel, 40-60 μ m, 120 μ m, Order No.: sw-5231 - 04 0-sp) detection wavelength 220 nm; 254 nm mobile phase solvent A: water; solvent B: acetonitrile flow rate 30 ml / min injection volume 5 ml elution gradient time (min) solvent B (%) 0 52 58 34 10 34 18 90 27 90 Figure 3 Flash preparation map of anthocyanin crude extract The flash preparation map of anthocyanin crude extract is shown in Figure 3 The collected target product is freeze-dried to obtain dry powder, and then it is dissolved for HPLC analysis, the results are shown in Figure 4 Compared with the HPLC analysis Atlas of the samples before and after flash purification preparation (Fig 2 and Fig 4), it can be seen that the purity of the effective component of the crude anthocyanin extract is increased from 84.2% to 97.2%, and the water-soluble impurities in the crude extract are effectively removed, and the obtained target product can be used for the next biological activity experiment and other purposes Figure 4 The HPLC analysis Atlas of anthocyanin crude extract after flash purification has expanded our thinking for future application and development through this flash purification attempt Santai technology will continue to be committed to the development of natural product separation and purification scheme, and provide more and better separation and purification solutions for customers To learn more about sepabean Gamma For detailed specification information of machine or ordering information of flash column, please visit CBG online store: https://store.chembergo.com/