Study On The Separation, Analysis And Anti-Inflammatory Activity Of Thlaspi Arvense L. Flavonoids

To clarify the structures and bioactivities of Thlaspi arvense L. flavonoids, we carried out further isolation, analysis and anti-inflammatory activity investigation. We described the isolation and identification of seven flavonoid glycosides and their in vivo anti-inflammatory activity in the present paper.1. The isolation and identification of Thlaspi arvense L. flavonoidsA new flavonoids compound, was isolated from Thlaspi arvense L. and identified as 5, 7-dihydroxy-flavone-4’-(6 "-O-β-D-glucoside)-O-β-D-glucoside (1). In addition, seven known flavonoids were also isolated:isosaponarin (2), isoorientin (3), vitexin (4), isovitexin (5), luteolin-7-O-β-D-glucoside (6), apigenin-7-O-β-D-glucoside (7) and 8-methoxy isovitexin (8). The structure of the isolated compounds was performed by chromatographic, chemical and spectroscopic methods. Compounds (2), (7) and (8) isovitexin were isolated from the genus Thlaspi for the first time.2. Simultaneous determination of seven flavonoids in Thlaspi arvense L. by RP-HPLCObjective:To establish a method for the simultaneous determination of 5,7-dihydroxy-flavone-4’-(6"-O-β-D-glucoside)-O-β-D-glucoside, isosaponarin, isoorientin, vitexin, isovitexin,luteolin-7-O-β-D-glucoside and apigenin-7-O-β-D-glucoside which were isolated from Thlaspi arvense L. Methods:The separation was performed on a Agilent Eclipse Plus C18 column (4.6×150mm,3.5μm) with a gradient elution composed of acetonitrile and 0.04% acetic acid aqueous solution. The column temperature was set at 30℃ and flow rate was 1.0 mL·min-1. The detective wavelength was set at 273 nm. Results:The content of 5,7-dihydroxy -flavone -4’-(6"-O-β-D-glucoside)-O-β-D-glucoside, isosaponarin, isoorientin, vitexin, isovitexin, luteolin-7-O-β-D-glucoside and apigenin-7-O-p-D-glucoside showed a good linearity in the range of 5.67～56.7 μg·mL-1 (r=0.9997),10.92～109.2 μg·mL-1 (r=0.9994), 5.76～57.6 μg·mL－1 (r=0.9998),4.92～49.2 μg·mL－1 (r=0.9998),13.95～139.5 μg·mL－1 (r=0.9999),5.25～52.5 μg·mL－1 (r=0.9998),5.22～52.2 μg·mL－1 (r=0.9997), respectively. The average recoveries (n=3) of seven flavonoids were 97%～101%　and all RSDs were less than 2.7%.3. Anti-inflammatory activity researchCompared with the aspirin and dexamethasone, anti-Inflammatory activities of seven flavonoids were evaluated using the murine models.The result showed that, isoorientin, vitexin, isovitexin and apigenin-7-O-p-D-glucoside had no significant anti-inflammatory effects on mice of xylene induced ear edema. Compared with blank,5,7-dihydroxy-flavone-4’-(6"-O-β-D-glucoside)-O-β-D-glucoside and isosaponarin aggravated the inflammatory state. Luteolin-7-O-β-D-glucoside had obviously anti-inflammatory effects (P<0.05).5,7-dihydroxy-flavon-4’-(6"-O-β-D-glucoside)-O-β-D-glucoside, isosaponarin, isovitexin and luteolin-7-O-β-D-glucoside had marked anti-inflammatory ability on mice of carrageenan induced paw edema. Isoorientin, vitexin and apigenin-7-O-β-D-glucoside had no significant anti-inflammatory effects.