1、 Yangtze River Pharmaceutical Group 扬子江药业集团有限公司1ReportProject: Polygoni orientalis fructusCustomer name and address: Yuan Ye / NKI-TCMProject objective: Evaluate TLC methods for Polygoni orientalis fructusDate: 8th February 2011Laboratory: NKI-TCMApproved by: Dr. Lu Zheng and Dr. Ting XianAimEstabli
2、shment of TLC methods for determination of Taxifolin in Polygoni orientalis fructus 1. General experimental detailsTable 1 Samples and standards identificationSample numberSample description Source or Supplier BatchS001 Polygoni orientalis fructusAnguo ginseng and antlerlimited company, Anguo city,
3、Hebei provinceL201010S002 Polygoni orientalis fructusAnguo ginseng and antler limited company, Anguo city, Hebei provinceJ201010S003 Polygoni orientalis fructusBeijing Tongrentang pharmacy, Beijing20101112S004 Polygoni orientalis fructusBeijing Yongantang pharmacy, Beijing20101118S005 Polygoni orien
4、talis fructusLotus pond TCM wholesale market, Chengdu city, Sichuan province201011WU HongweiS006 Polygoni orientalis fructusZhongyiyuan pharmacy, Shenyang city, Liaoning province101201S007 Polygoni orientalis fructusTaizhou Gaogang TCM Co., LTD, Taizhou City, Jiangsu province030111S008 Polygoni orie
5、ntalis fructusJilin city, Jilin province 20101121S009 Polygoni orientalis fructusChina Nepstar Chin Drug strore LTD,Siping city, Jilin province091227S010 Polygoni orientalis Northeast pharmacy, Shenyang city, Liaoning 201011Yangtze River Pharmaceutical Group 扬子江药业集团有限公司2fructus provinceTable 2 Stand
6、ardCAS No. Standard description Source batch480-18-2 Taxifolin NIFDC 111816-201001491-54-3 Kaempferol NIFDC 110861-200808Table 3 ChemicalsName Manufacturer BatchMethanol West gansu chemical Co. Ltd110604-1Trichloromethane Sinopharm chemical reagent Co. ,Ltd100707-1Acetone West gansu chemical Co. Ltd
7、10000492Glacial acetic acid West gansu chemical Co. Ltd101020-1Anhydrous formic acid West gansu chemical Co. Ltd100707-1Diphenylboric acid aminoethyl esterAcros Organics A02801Macrogol West gansu chemical Co. Ltd100130-1Table 4 PlatesType ManufacturerHPTLC Si 60 , 10 x 10 cm MerckTable 5 EquipmentsN
8、ame, article ManufacturerThermostatwaterbath Shanghai boxun Co. LtdThermoelectricthermostatdryingbox Shanghai boxun Co. LtdCamera PatriotUltraviolet analyzes Gongyi instrument equipment Co. LtdAnalytical balance Mettler-Toledo2. Samples and standard preparationYangtze River Pharmaceutical Group 扬子江药
9、业集团有限公司3Test solution: Heat 2 g of the powdered drug in 5 ml of methanol R under a reflux condenser for 10 min. Cool, then filtered. Use the filtrate as the test solution.Reference solution: Dissolve 1 mg of taxifolin R in 2 ml of methanol R. Dissolve 1 mg of kaempferol R in 10 ml of methanol R.3. R
10、esults (plates)ApplicationInstrument: Drummond scientific CO USABand length: 8 mm Distance between tracks: 7 mmApplication position: X: 20 mm Y: 10 mmChromatographyPlate: HPTLC glass 20X10 cm, Si 60 Chamber: ADC 2Plate conditioning: at 50% humiditySaturation: 60 min with filter paperDeveloping dista
11、nce from application position/lower edge: 80 mm.Developing solvent: Trichloromethane R, Acetone R, Glacial acetic acid R, anhydrous Formic acid R (20:1:2:2 V/V/V/V).Plate drying: 10 min, 105 .Detection: spray the warm plate with a 10 g/L solution of diphenylboric acid aminoethyl of ester R in methan
12、ol R, and then with the same volume of a 50 g/L solution of macrogol 400 R in methanol R, allow drying in air for about 30 min and evaluating under UV 365 nm. Top of the plateA green zoneA red zoneA pale blue fluorescent zoneA diffuse green fluorescent zoneA blue fluorescent zone A green fluorescent zone or a blue fluorescent zone Yangtze River Pharmaceutical Group 扬子江药业集团有限公司4A orange zoneA green fluorescent zoneA blue fluorescent zoneA orange zoneA orange zoneA blue fluorescent zoneReference solution Test solution
