Studies On Chemical Constituents Of S. Tonkinensis And S. Flavescens,and Transformation Of Saikosaponin A, D In Different Extracting Method By Using UPLC-ESI-QTof

"Shandougen" is the dried roots and rhizomes of Sophora tonkinensis Gapnep., which has the effects of clearing away heat, reducing swelling and relieving sore throat."Kushen" is the dried roots of Sophora flavescens Ait which is used to clean heat, dry humidity and as a diuretic. Traditional chemical separation research shows that the chemical compositions in those two plants are similar, and it is hard to explain the differences of clinical applications from the view of material foundations. Firstly, UPLC-PDA method was used to screen the representative herbs of Radix Sophorae Tonkinensis and the Radix Sophorae flavescentis, then UPLC-ESI-QTof method which was of highly sensitivity was employed to systematically study chemical components in the roots of Radix Sophorae Tonkinensis and Radix Sophorae flavescentis.The analysis was performed on an Acquity UPLC BEH C18column (2.1mm×50mm,1.7μm), using5mmol/L ammonium formate in water (B) and acetonitrile (A) as mobile phase, and quadrupole-time of flight (Q-Tof) mass spectrometry as the detector with positive ion scanning mode, to set up the UPLC-ESI-QTof analysis method of alkaloids in the root of Radix Sophorae Tonkinensis and Radix Sophorae flavescentis. The results shown that the alkaloid compositions which were in the root of the two herbs were similar, mainly contained5α,9α-Dihydroxy matrine, oxymatrine, oxysophocarpine and matrine. The content of alkaloids differ widely, which including matrine, Sophoranhol N-oxide, oxymatrine, oxysophocarpine, sophoridine, N-Methylcytisine and sophocarpine.For the non-alkaloid compositions, the analysis was performed on an Acquity UPLC BEH C18column (2.1mm×50mm,1.7μm) by using0.01%formic Acid in water (B) and acetonitrile (A) mobile phase with the negative ion scanning mode. The flavonoids and saponins are the main components in the root of Radix Sophorae Tonkinensis, such as sophoradin Ⅰ, sophoranochromene Ⅶ, subproside Ⅱ, isoanhydroicarition, while kushenol E, kurarinone, kushenol Ⅰ, isokurarinone are characteristic components which existed in the root of Sophora flavescens Ait.To further identify the different non-alkaloid constituents between the root of the Radix Sophorae Tonkinensis and the Radix Sophorae flavescentis. Extraction and separation of non-alkaloid constituents from the Radix Sophorae Tonkinensis was undergone. By employing various modern chromatographic techniques,11compounds (S1～S13) were isolated from non-alkaloid part and their structures were elucidated by spectroscopic data including UV,1HNMR,13CNMR, ESI-MS. There were5flavonoids and flavonoid glycoside including3,4’-dihydroxy-7-methoxy-flavones(S6),7,4’-dihydroxy flavones(S7), trifolirhizin(S8), maaekiain(S13) isoquercitrin(S11);4sterols and steroid saponins:β-sitosterol (S3), stigmasterol (S5) daucosterol (S9), stigmasterol-3-O-β-D-glucopyranoside (S10);1aliphatic ester:ethyl linolkeate (S1);2aryl acid esters:3-(3-hydroxy-4-methoxyphenyl)-2-propenoic acid docosyl ester (S2),2,3-dihydroxy-2-[(4-hydroxyphenyl)methyl]-butanedioic acid-4-ethyl ester(S12), and1triterpene:lupeol (S4). Among the compounds, S1, S2, S12were isolated from this plant for the first time.In addition, we studied the transformation of saikosaponin A and saikosaponin D in different extracting methods by UPLC-ESI-QTof. A new UPLC-ESI-QTof method has been developed for profiling and quantitating the transformation of saikosaponin A and saikosaponin D by different extracting methods. The separation was performed on a Acquity UPLC BEH C18column(2.1mm×50mm,1.7μm). A mobile phase consisting of0.01%formic Acid in water (B) and acetonitrile (A) was used for the separation. The flow rate was kept constant at0.45mL/min with gradient elution. Quadrupole-time of flight (Q-Tof) mass spectrometry was used as the detector. Twelve samples of radix bupleuri, which obtained by different extraction methods, were analyzed by electron pray ionization mass spectrometry(ESI-MS) with negative ion mode. The result of transformation between saikosaponin A and saikosaponin D extracted in different methods were derived:saikosaponin A and saikosaponin D were hydrolyzed into saikosaponin B1and saikosaponin B2under acidic condition at room temperature. The glycosidic bond was hydrolyzed to yield sapogenin under the acid condition and being heated, while it could be stabilized under neutral and alkaline condition, which also provided a basis for the quality control of Radix bupleuri medicinal materials and its Chinese patent medicines.