德国General Declarations and Spe

1、Zentralinstitut ArzneimittelforschungPart of this documentation is CD-No. 0001 including all data and statistical evaluation First version: 0.1Current version: 1.0Created on 07.01.2002 Created by Dr. Udo Söker Created by Dr. Udo Söker Last print: 08.02.02 Musterdoku_neu_SAMPLE M:AustauschJ

2、oerg.WittigCd_Musterdokumentation_26.02.2002Musterdoku_neu_SAMPLE.docDocumentation SpiraeosideZentralinstitut ArzneimittelforschungGeneral InformationExperimental: U. Söker, PhD, chemistMelanie Much, technicianStudy director: M. Veit, PhD, pharmacistLocation of laboratories and archives:Kranzwe

3、iherweg 1053489 SINZIGGERMANYVersion 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 2 of 27Zentralinstitut ArzneimittelforschungSubstance:Batch No:Manufacturer:Amount: Spiraeoside SP-01-2/4 Sinzig, Germany Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 3 o

4、f 27Zentralinstitut ArzneimittelforschungIndex11.11.21.31.422.12.22.42.52.62.72.8NMR-Spectral Analysis16 16 19 21 22 24 25 27Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 4 of 27Zentralinstitut Arzneimittelforschung1 General Declarations and SpecificationsSpiraeoside:C21HE

5、mpirical formula: No data available.Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 5 of 27Zentralinstitut Arzneimittelforschung1.3 ManufacturingProcedureThe substance was isolated from the dried bulb coats of Allium cepaEtOH/H2O.1.4 Isolation of the Substanceevaporated in v

6、acuoin vacuo. 2Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 6 of 27Zentralinstitut Arzneimittelforschung2Identity and Purity2.1 NMR-Spectral AnalysisThe NMR spectra of spiraeoside were recorded using CD3SOCD3 and CDInc. (USA).Instrument: Bruker AM360, Detection: 1Temperat

7、ure: 303 KProbe head:NMR tube:31H)11H NMR coupling pattern as (2.1 , 5.6 Hz). It must be mentioned that in some cases, e.g. the anisomeric proton H-1” as well as the protons H-2”, H-3”, and H-4”, the presence of higher order spin Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside pa

8、ge 7 of 27Zentralinstitut Arzneimittelforschungsystems requires labeling of the signals as multiplets and analysis of the active Nat. Prod. 2000,63 (6), 834-838. The sample was measured in CD3CD3SOCD3Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 8 of 27Zentralinstitut Arzn

9、eimittelforschung1H NMR data (solvent CD3SOCD3)Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 9 of 27Zentralinstitut Arzneimittelforschung13C NMR Spectral Analysis (spectrum see appendix):Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 10 of 27Zentr

10、alinstitut Arzneimittelforschung2.2 Mass Spectral AnalysisVersion 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 11 of 27Zentralinstitut Arzneimittelforschung2.3 Infrared Spectral AnalysisVersion 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 12 of 27Zentralinstit

11、ut Arzneimittelforschung2.5 Melting pointVersion 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 13 of 27Zentralinstitut ArzneimittelforschungVersion 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 14 of 27Zentralinstitut Arzneimittelforschung2.8 X-Ray Crystallograp

12、hyVersion 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 15 of 27Zentralinstitut Arzneimittelforschung3 Assayindependent methods (100% peak area method).Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 16 of 27Zentralinstitut ArzneimittelforschungFlow : 0.25

13、 ml min-1Injection volume :Time of analysis: 20 µL 55 minDetection wavelength : 254 nmAnalyte solutionBlank value :Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 17 of 27Zentralinstitut ArzneimittelforschungResults of the quantitative analysis:conditions was carried ou

14、t and the standard deviation was calculated.Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 18 of 27Zentralinstitut ArzneimittelforschungSelectivityby comparison with a second independent method.Sensitivity-1detected at 0.1 % level.Linearityi. d. 50 µm; length 50 cm, ef

15、f. 46.5 vm254 nm Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 19 of 27Zentralinstitut ArzneimittelforschungAnalyte solution: About 10 mg (9.81 mg) spiraeoside, exactly weighted, wasVersion 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 20 of 27Zentralins

16、titut ArzneimittelforschungMethod ValidationAccuracyassay.PrecisionSelectivitymg mL-1 proved,acetonitrile a second internal standard (n-propyl acetate, ISTD2) was used. Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 21 of 27Zentralinstitut ArzneimittelforschungThe validatio

17、n of the method was performed for 19 of the most common solvents:Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 22 of 27Zentralinstitut ArzneimittelforschungResults of the quantitative analysis:Version 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 23 of 2

18、7Zentralinstitut ArzneimittelforschungVersion 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 24 of 27Zentralinstitut Arzneimittelforschung4 Certificate of AnalysisVersion 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 25 of 27Zentralinstitut ArzneimittelforschungV

19、ersion 1.0 Dr. Udo Söker 08.02.2002 Documentation Spiraeoside page 26 of 27Zentralinstitut Arzneimittelforschung5 Appendix1H and 13C NMR spectra 2 pagesMass spectra 1 pageInfrared spectrum 1 Thin-Layer Chromatogram 1 Ultraviolet spectrum X-Ray Data HPLC assay CE assay Version 1.0 Dr. Udo Sö

20、;ker 08.02.2002 Documentation Spiraeoside page 27 of 27ESI MS Spectrum of SpiraeosideSingle MSS#:2-4RT:0.03-0.09AV:3NL:1.25E6T:+ c Full ms 130.00 - 1000.00x510095908580757065Relative Abundance605550454035302520151050150149.3176.8205.3227.3244.5200250279.2300304.5353.3350m/z379.3400431.1487.3438.6450

21、500509.3566.9550582.8600466.4465.3Analyte solution: about 10 mg spiraeoside in 10 ml methanol· white light· Naturstoffreagenz (A) and Macrogol 400 solution · l = 365 nmDatum: 19.07.100Zeit: 16:09:07 0,9060,850,800,750,700,650,600,550,50A 0,450,400,350,300,250,200,150,100,050,001200,02

22、50300350400nm450500550600650,0365,42253,02Beschreibung: Spiraeosid, 9,55 µg/mlDateiname: C:UVWINLABDATAMM-0494.SP Gemessen am: 06.07.00 11:57:48Spektrometer: Lambda 40 PDatenintervall: 1,0000 nmgelöst in Methanol, Gradient gradeRegistriergeschwindigkeit: 120,00 nm/minSpaltbreite: 1,0000 nm

23、Glättungsintervall: 0,00 nmTable 1. Crystal data and structure refinement for FRO1265. Identification code FRO1265 Empirical formula C21 H24 O14 Formula weight 500.40 Temperature 198(2) K Wavelength 0.71073 Å Crystal system, space group orthorhombic, P212121 (No. 19) Unit cell dimensions a

24、 = 4.412(1) Å b = 13.344(1) Å c = 35.985(1) Å Volume 2118.6(5) Å Z, Calculated density 4, 1.569 Mg/m Absorption coefficient 0.134 mm F(000) 1048 Crystal size 0.20 x 0.10 x 0.05 mm Theta range for data collection 1.63 to 27.47°. Limiting indices -5<=h<=3, -17<=k<=

25、10, -30<=l<=46 Reflections collected / unique 6499 / 4583 R(int) = 0.0309 Completeness to theta = 27.47 99.2 % Max. and min. transmission 0.9933 and 0.9736 Refinement method Full-matrix least-squares on F Data / restraints / parameters 4583 / 6 / 336 Goodness-of-fit on F21.072Final R indices I

26、>2 s(I) R1 = 0.0676, wRR indices (all data) R1 = 0.1069, wR Absolute structure parameter 0.0(15) Largest diff. peak and hole 0.333 and -0.285 eÅ 3 3 -1 2 2 = 0.1191 2 = 0.1340 -3Z e n t r a l i n s t i t u t A r z n e i m i t t e l f o r s c h u n g G m b HKranzweiherweg 10 D-53489 Sinzig/Rh

27、einValidation of NPRSReference StandardSpiräosidSpiräosid; 0,107 mg/mlSampleName: Spiräosid MM0505 Injection: 2 Vial: 3Injection Vol.: 10,00 ulRun Time: 60,00 MinutesDate Acquired: 17.07.00 22:48:03 Method Set: NPRS 2Processing Method: Spiräosid_ZA_PDA_254_PMColumn: MZ Kromasil 100 C18, 250 x 4 mm, 5 µmPre-Column: MZ Kromasil 100 C18, 10 x 4 mm, 5 µm0,450,400,350,300,25AU0,200,150,100,000,002,004,006,008,0010,0012,0014,0016,00Minutes18,0020,0022,0024,0026,0028,0030,00Channel Description PDA 254,0 nmSampleName Spiräosid MM0505 Injection 2