| Sophorae Radix (SR) is collected from the root of Sophora flavescens Ait. belonging to the genus Sophora. SR has been widely used as a traditional Chinese medicine with the property of clearing heat away and eliminating dampness, facilitating urination and removing the bacteria or fungi. It is bitter in flavor and cold in nature. With the increasingly extensive study on its chemical constituents, over 39 quinolizidine alkaloids and 122 flavonoids have been isolated and identified from it. Among them, quinolizidine alkaloids and flavonoids are considered as active ingredients.Due to its definite clinical effects, it was widely used in clinics and TCM industries. Now, over 166 TCM finished products contained SF were manufactured and well reputation in market. It was also widely used as raw materials for production of veterinary drugs, biopesticides and commodities. With the hungry requirement of industries, its wild resource was rapidly decreased.As a perennial herb, the root of SF as medicinal part, need at least 3years to harvest. In these growing period, a large amount of its aerial parts, such as seed, leaf, stem and flower, was lavished as agricultural wastes. To comprehensively utilize these resources, we focus these rich resources to investigate their active main ingredients and compositions. Part 1 Constituents rapid identification by UPLC-qTOF-MS/MSA qualitative study on the flavonoids and alkaloids present in different organs of SF was conducted by UPLC-qTOF-MS/MS. Based on the comparisons of the retention times, UV spectra with those of the reference substances and the accurate quasi-molecular ion as well as fragmentation rules,156 compounds were identified, including 28 alkaloids,98 flavonoids,29 triterpene saponins and 1 lignin. Among them,53 compounds were first-detected in this plant, and 22 compounds could be new ones.The results showed that different organs contain different compounds and types. The root not only contains alkaloids but also flavones include flavanone, flavanol and isoflavone glycosides. The stem and leaf mainly contain isoflavone glycosides, while flower contains flavonoid glycosides. A lot of triterpene glycosides were detected in the flower and seed also.Part 2 Quantitative analysis for different organsThe quantitative determination of oxymatrine, matrine, oxysophocarpine, sophocarpine and N-methylcytisine were conducted by HPLC. The total flavonoids were measured by ultraviolet spectrophotometry at 260 nm, using genistin as a reference substance. The total alkaloids were determined using acid dye colorimetry at 417 nm, using oxymatrine as a reference substance. The results indicated that the contents of flavonoids and alkaloids were different in the different organs. The highest content of flavonoids is in root, followed by flower and leaf. The content of alkaloids in root and seed were more than those in the other organs. Oxymatrine as the greatest amount of alkaloid constituent was found in all the organs. Part 3 Quality standard investigation for root extract of Sophora flavescensAs a part of the project for the Chinese Pharmacopoeia (2015 Edition), the quality standard of Sophora flavescens root extract was investigated and established. According to the methods described in the Appendix of Chinese Pharmacopoeia (2010 Edition), the water and ash inspections were carried out. The marker components trifolirhizin, sophoraflavanone G, oxymatrine and oxysophocarpine in the samples were identified by qualitative TLC. The determination of oxymatrine, matrine, oxysophocarpine and sophocarpine was conducted by HPLC and the total flavonoids were measured by ultraviolet spectrophotometry, using sophoraflavanone G as a reference substance. The results indicated the spots on the plate were clear with good resolution and the contents of oxymatrine, matrine, oxysophocarpine and sophocarpine in the 13 batches of the samples were 3.87%-11.06%,0.97%-4.33%, 1.30%-2.59%and 0.26%-1.14%, respectively. The total flavonoids in the 13 batches of the samples were 3.88%-7.93%. In the study, the validated methods were reproducible and the established quality standard was feasible, which could be used for the quality control of Sophora flavescens root extract and related preparations. |
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