CTD Open Cefoperazone

HARBINHARBINHARBINHARBIN PHARMACEUTICALPHARMACEUTICALPHARMACEUTICALPHARMACEUTICAL GROUPGROUPGROUPGROUP CO.,CO.,CO.,CO., LTDLTDLTDLTD GeneralGeneralGeneralGeneral Pharm.Pharm.Pharm.Pharm. FactoryFactoryFactoryFactory

CTDCTDCTDCTD ModuleModuleModuleModule 3.2.S3.2.S3.2.S3.2.S forforforfor SterileSterileSterileSterile CefoperazoneCefoperazoneCefoperazoneCefoperazone SodiumSodiumSodiumSodium

THETHETHETHE COMMONCOMMONCOMMONCOMMON TECHNICALTECHNICALTECHNICALTECHNICAL DOCUMENTDOCUMENTDOCUMENTDOCUMENT 3.2.S3.2.S3.2.S3.2.S FORFORFORFOR THETHETHETHEREGISTRATIONREGISTRATIONREGISTRATIONREGISTRATION OFOFOFOF PHARMACEUTICALSPHARMACEUTICALSPHARMACEUTICALSPHARMACEUTICALS FORFORFORFOR HUMANHUMANHUMANHUMAN USE:USE:USE:USE:

SterileSterileSterileSterile CefoperazoneCefoperazoneCefoperazoneCefoperazone SodiumSodiumSodiumSodium

OpenOpenOpenOpen partpartpartpart

HARBINHARBINHARBINHARBIN PHARMACEUTICALPHARMACEUTICALPHARMACEUTICALPHARMACEUTICAL GROUPGROUPGROUPGROUP CO.,CO.,CO.,CO., LTD.LTD.LTD.LTD.GeneralGeneralGeneralGeneral Pharma.Pharma.Pharma.Pharma. FactoryFactoryFactoryFactory

2020202011111111



3.13.13.13.1 TABLETABLETABLETABLE OFOFOFOF CONTENTSCONTENTSCONTENTSCONTENTS OFOFOFOF MODULEMODULEMODULEMODULE 33333.23.23.23.2 BODYBODYBODYBODY OFOFOFOF DATADATADATADATA3.2.S DRUG SUBSTANCE3.2.S.1 General Information3.2.S.1.1 Nomenclature3.2.S.1.2 Structure3.2.S.1.3 General Properties3.2.S.2 Manufacture3.2.S.2.1 Manufacturer(s)3.2.S.2.2 Description of Manufacturing Process and Process Controls3.2.S.2.3 Control of Materials3.2.S.2.4 Control of Critical Steps and Intermediates3.2.S.2.5 Process Validation and/or Evaluation3.2.S.2.6 Manufacturing Process Development3.2.S.3 Characterization3.2.S.3.1 Elucidation of Structure and other Characteristics3.2.S.3.2 Impurities3.2.S.4 Control of Drug Substance3.2.S.4.1 Specification3.2.S.4.2 Analytical Procedures3.2.S.4.3 Validation of Analytical Procedures3.2.S.4.4 Batch Analyses3.2.S.4.5 Justification of Specification3.2.S.5 Reference Standards or Materials3.2.S.6 Container Closure System3.2.S.7 Stability3.2.S.7.1 Stability Summary and Conclusions3.2.S.7.2 Post-approval Stability Protocol and Stability Commitment3.2.S.7.3 Stability Data



3.23.23.23.2 BODYBODYBODYBODY OFOFOFOF DATADATADATADATA3.2.S3.2.S3.2.S3.2.S DRUGDRUGDRUGDRUG SUBSTANCESUBSTANCESUBSTANCESUBSTANCE3.2.S.13.2.S.13.2.S.13.2.S.1 GeneralGeneralGeneralGeneral InformationInformationInformationInformation



3.2.S.1.13.2.S.1.13.2.S.1.13.2.S.1.1 NomenclatureNomenclatureNomenclatureNomenclature

GeneralGeneralGeneralGeneral remarkremarkremarkremarkThis Drug Master File has been compiled to provide the necessary information for the evaluationof suitability of the bulk substance of CEFOPERAZONE SODIUMmanufactured by HarbinPharmaceutical Group CoLtd

It is used as active substance for medicinal products(dosage forms)which are blended withsulbactam sodium generally. The structure of the document represents the section 3.2.S of themodule 3 of the Common Technical Document(CTD)Thusit is appropriate to be integrated in thequality part of an application for marketing authorization

The content of this Active Substance Master File is property of Harbin Pharmaceutical Group CoLtd. and thereforeit is confidentialThis document should not be disclosed no matter if in wholeor in part Not any parties other than customers and government employees required forregulatory purposes are authorized to have access to its content

The manufacture of cefoperazone sodium is performed according to the presented dossier and totheGood Manufacturing Practice of the Peoples Republic of China.

On request of relevant authorities and customers the inspection regarding the manufacture ofcefoperazone sodium is possible

Cefoperazone sodium is an antibiotic compound of the third generation of cephaIosporins Itshows a high resistance against -lactamase And because sulbactam can strengthen theantibacterial activity of it, cefoperazone sodium has been often blended with sulbactam sodium tocomply the requirements of customers.



The active ingredient is commonly known under the name

CefoCefoCefoCefoperazoneperazoneperazoneperazone sodiumsodiumsodiumsodium

This term is used in the compendial title of the monograph of the European Pharmacopoeiapublished in its current editionThe cefoperazone sodium produced by Harbin PharmaceuticalGroup CoLtdNangang DistrictChina corresponds to the quality standard of this monograph

ChemicaIChemicaIChemicaIChemicaI namesnamesnamesnames

SodiumSodium(6R,7R)-7-[[(2R)-2-[[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonyl]amino]-2-(4-hydroxyphenyl)acetyl]amino]-3-[[(1-methyl-1H-tetrazol-5-yl)sulphanyl]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate.(EP 6.0)

5-Thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid,7-[[[[(4-ethyl-2,3-dioxo-1-piperazinyl)carbonyl]amino](4-hydroxyphenyl)acetyl]amino]-3-[[(1-methyl

-1H-tetrazol-5-yl)thio]methyl]-8-oxo-, monosodium salt, [6R-[6 ,7 (R*)]]-.

(USP 32)

Sodium(6R,7R)-7-[(R)-2-(4-ethyl-2,3-dioxo-1-piperazinecarboxamido)-2-(p-hydroxyphenyl)acetamido-3-[[(1-methyl-H-tetrazol-5-yl)thio]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate(USP 32)

CAS-Registry-NumberCAS-Registry-NumberCAS-Registry-NumberCAS-Registry-Number

[62893-20-3]



3.2.S.1.23.2.S.1.23.2.S.1.23.2.S.1.2 StructureStructureStructureStructure

The chemical structure of cefoperazone sodium is exhibited below.

CefoCefoCefoCefoperazoneperazoneperazoneperazone sodiumsodiumsodiumsodium

MolecularMolecularMolecularMolecular formulaformulaformulaformula

C25H26N9NaO8S2

RelativeRelativeRelativeRelative molecularmolecularmolecularmolecular massmassmassmass

668



3.2.S.1.33.2.S.1.33.2.S.1.33.2.S.1.3 GeneralGeneralGeneralGeneral PropertiesPropertiesPropertiesProperties

AppearanceAppearanceAppearanceAppearanceA white or slightly yellow powder

HygroscopicityHygroscopicityHygroscopicityHygroscopicityHygroscopic

pkpkpkpkaaaa-value-value-value-valueThe pH of the solution is 4.5 to 6.5.

InfraredInfraredInfraredInfrared absorptionabsorptionabsorptionabsorption spectrophotometryspectrophotometryspectrophotometryspectrophotometryExamine by infrared absorption spectrophotometry (2.2.24), comparing with the Ph. Eur. referencespectrum of cefoperazone sodium.

SolubilitySolubilitySolubilitySolubilityFreely soluble, in water, soluble in methanol, slightly soluble in alcohol.

PolymorphismPolymorphismPolymorphismPolymorphismIf crystalline, it shows polymorphism.



3.2.S.23.2.S.23.2.S.23.2.S.2 ManufactureManufactureManufactureManufacture



3.2.S.2.13.2.S.2.13.2.S.2.13.2.S.2.1 Manufacturer(s)Manufacturer(s)Manufacturer(s)Manufacturer(s)

The name and address of the manufacturing site of the active ingredient manufacturer is:

HARBINHARBINHARBINHARBIN PHARMACEUTICALPHARMACEUTICALPHARMACEUTICALPHARMACEUTICAL GROUPGROUPGROUPGROUP CO.,CO.,CO.,CO., LTD.LTD.LTD.LTD. GeneralGeneralGeneralGeneral Pharm.Pharm.Pharm.Pharm. FactoryFactoryFactoryFactory

No. 109 Xuefu RoadNangang Dist.Harbin, Peoples Republic of China

Tel: 0086-0451-86665134Fax: 0086-0451-86662959E-mail: [email protected]



3.2.S.2.23.2.S.2.23.2.S.2.23.2.S.2.2 DescriptionDescriptionDescriptionDescription ofofofof ManufacturingManufacturingManufacturingManufacturing ProcessProcessProcessProcess andandandand ProcessProcessProcessProcess ControlsControlsControlsControls

A.A.A.A. SyntheticSyntheticSyntheticSynthetic routerouterouteroute

CefoperazoneCefoperazoneCefoperazoneCefoperazone acidacidacidacid

PreparationPreparationPreparationPreparation ofofofof TZATZATZATZA

DissolveDissolveDissolveDissolve ofofofof TZATZATZATZA

NN

NS

N

CH3S

NH2N

OCOOSi(CH3)3

CH2CH3CN

BSACH2

COOHO

H2NN

S CH3

NNS

NN

AcylAcylAcylAcyl chloridechloridechloridechloride reactionreactionreactionreaction

AcylationAcylationAcylationAcylation

COCl

OH

CHNHCO

C2H5

O O

N N

CH2COOSi(CH3)3

O

H2NN

S CH3

NNS

NN

+



CH3C

OC NH CH

OH

O O

NC2H5 N

COOSi(CH3)3

HHO

NN

NSCH2

S

O NN

N

hydrolysishydrolysishydrolysishydrolysis

NC2H5 N

OO

OH

CHNHCO

CCH3

NN

NO

S

CH2 SN

NN

O HH

COOSi(CH3)3

H2O

HHO

NN

NSCH2

S

O NN

N

CH3C

OC NH CH

OH

O O

NC2H5 N

COOH

CefoperazoneCefoperazoneCefoperazoneCefoperazone SodiumSodiumSodiumSodium



3.2.S.2.33.2.S.2.33.2.S.2.33.2.S.2.3 ControlControlControlControl ofofofof MaterialsMaterialsMaterialsMaterials

In the following starting materials will be presented which are mentioned in the description of themanufacturing process in section 3.2.S.2.2:



AAAAcetonitrilecetonitrilecetonitrilecetonitrile

MolecularMolecularMolecularMolecular formulaformulaformulaformula:::: CH3CNMolecularMolecularMolecularMolecular weightweightweightweight:::: 41.05



BFBFBFBF3333-Acetonitrile-Acetonitrile-Acetonitrile-Acetonitrile

MolecularMolecularMolecularMolecular formulaformulaformulaformula:::: C 2H3NBF 3MolecularMolecularMolecularMolecular weightweightweightweight:::: 109Structure:Structure:Structure:Structure:



5-mercapto-1-methyltetrazole5-mercapto-1-methyltetrazole5-mercapto-1-methyltetrazole5-mercapto-1-methyltetrazole

MolecularMolecularMolecularMolecular formulaformulaformulaformula:::: C2H4N4SMolecularMolecularMolecularMolecular weightweightweightweight:::: 116.14Structure:Structure:Structure:Structure:



7-ACA7-ACA7-ACA7-ACA

MolecularMolecularMolecularMolecular formulaformulaformulaformula:::: C 10H 12N 2O 5SMolecularMolecularMolecularMolecular weightweightweightweight:::: 272.27Structure:Structure:Structure:Structure:



AmmoniaAmmoniaAmmoniaAmmonia WaterWaterWaterWater (refined)(refined)(refined)(refined)

MolecularMolecularMolecularMolecular formula:formula:formula:formula: NH4OH or NH3 H2OMolecularMolecularMolecularMolecular weight:weight:weight:weight: 35.10



AcetoneAcetoneAcetoneAcetone

MolecularMolecularMolecularMolecular formulaformulaformulaformula: C3H6OMolecularMolecularMolecularMolecular weightweightweightweight: 58.08StructureStructureStructureStructure:



6%6%6%6% sulfuroussulfuroussulfuroussulfurous acidacidacidacid

MolecularMolecularMolecularMolecular formulaformulaformulaformula: H2SO3MolecularMolecularMolecularMolecular weightweightweightweight: 82.08



N,O-Bis(trimethylsilyl)acetamideN,O-Bis(trimethylsilyl)acetamideN,O-Bis(trimethylsilyl)acetamideN,O-Bis(trimethylsilyl)acetamide (BSA)(BSA)(BSA)(BSA)

MolecularMolecularMolecularMolecular formulaformulaformulaformula: C8H21NOSi2MolecularMolecularMolecularMolecular weightweightweightweight: 203.43StructureStructureStructureStructure:



DimethylacetamideDimethylacetamideDimethylacetamideDimethylacetamide ((((DMAC)DMAC)DMAC)DMAC)

MolecularMolecularMolecularMolecular formulaformulaformulaformula: CH3CON(CH3)2MolecularMolecularMolecularMolecular weightweightweightweight: 87.12StructureStructureStructureStructure:



PPPPhosphorushosphorushosphorushosphorus oxychlorideoxychlorideoxychlorideoxychloride

MolecularMolecularMolecularMolecular formulaformulaformulaformula: POCl3MolecularMolecularMolecularMolecular weightweightweightweight: 153.33



ActiveActiveActiveActive carboncarboncarboncarbon #767#767#767#767

MolecularMolecularMolecularMolecular formula:formula:formula:formula: CMolecularMolecularMolecularMolecular weight:weight:weight:weight: 12.01



SodiumSodiumSodiumSodium BicarbonateBicarbonateBicarbonateBicarbonate

MolecularMolecularMolecularMolecular formula:formula:formula:formula: NaHCO3MolecularMolecularMolecularMolecular weight:weight:weight:weight: 84.01



3.2.S.2.43.2.S.2.43.2.S.2.43.2.S.2.4 ControlsControlsControlsControls ofofofof CriticalCriticalCriticalCritical StepsStepsStepsSteps andandandand IntermediatesIntermediatesIntermediatesIntermediates

With reference to section 3.2.S.2.2 it is obvious from the description of the manufacturingprocess that during this process cefoperazone acid, as an isolated intermediate, isobtained. This cefoerazone acid is still nonsterile. For this reason a separateinprocess control of the key intermediate cefoperazone acid takes place and theanalytical tests are described. The reasonable inprocess specification ensures that thefinished active substance meets the specification.

PreparationPreparationPreparationPreparation ofofofof TZATZATZATZA

ProcessProcessProcessProcess ControlControlControlControl pointpointpointpoint SpecificationSpecificationSpecificationSpecification TreatmentTreatmentTreatmentTreatment

Condensationreaction

Temperature 121 Control thetemperature

Assay of 7-ACAafter reaction.

< 2.0 mg/ml

CrystallizationpH 3.1 ~ 3.3

Control usingammonia water

Temperature 91 Control thetemperature

CefoperazoneCefoperazoneCefoperazoneCefoperazone acidacidacidacid


Dissolve of TZA Temperature 20 ~23 Control using hot

waterAcyl chloridereaction

Temperature -25 ~ -30 Control using liquid

nitrogen

AcylationTemperature -302

Control using liquidnitrogen

Assay of TZA afteracylation.

< 2 %

Crystallization

Assay ofCefoperazone

side-chain acid < 3mg/ml after

crystallization.

< 3.0 mg/ml



CefoperazoneCefoperazoneCefoperazoneCefoperazone SodiumSodiumSodiumSodium


Forming sodiumsalt reaction

Temperature

pH

20 ~ 25

6.0 ~ 7.0

Control the temperatureusing ethylene glycol inmezzanine.Adjust using NaHCO3slowly.

Crystallization Temperature

How long the acetone dropping

26 2

For 4.5 hours

Control the temperatureusing ethylene glycol inmezzanine.

Centrifugalizationand washing

Turns when transfer crystallizationsolutionTurns when add washing solutionTurns when centrifugalization

600 50 rpm

600 50 rpm900 300 rpm

Adjust the turns permins.

Drying TemperatureWater

60 2 Hot water circulationEvacuate during drying



3.2.S.2.53.2.S.2.53.2.S.2.53.2.S.2.5 ProcessProcessProcessProcess ValidationValidationValidationValidation and/orand/orand/orand/or EvaluationEvaluationEvaluationEvaluation

Documentation on process validation is given in the Restricted Part of the ActiveSubstance Master File and is submitted directly to registration authorities on request.



3.2.S.2.63.2.S.2.63.2.S.2.63.2.S.2.6 ManufacturingManufacturingManufacturingManufacturing ProcessProcessProcessProcess DevelopmentDevelopmentDevelopmentDevelopment

Not applicable.



3.2.S.33.2.S.33.2.S.33.2.S.3 CharacterizationCharacterizationCharacterizationCharacterization



3.2.S.3.13.2.S.3.13.2.S.3.13.2.S.3.1 ElucidationElucidationElucidationElucidation ofofofof StructureStructureStructureStructure andandandand otherotherotherother CharacteristicsCharacteristicsCharacteristicsCharacteristicsChemicalChemicalChemicalChemical structurestructurestructurestructure analysisanalysisanalysisanalysis ofofofof CefoperazoneCefoperazoneCefoperazoneCefoperazone sodiumsodiumsodiumsodium

I.I.I.I. NameNameNameName ofofofof compoundcompoundcompoundcompound

Drug name (English): Cefoperazone Sodium

Chemical name (English):

Sodium

(6R,7R)-3-[[(1-methyl-H-tetrazol-5-yl)thio]methyl]-7-[(R)-2-(4-ethyl-2,3-dioxo-1-piperazinecarboxa

mido)-2-(phydroxy phenyl)acetamido]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-carboxylate

II.II.II.II. MolecularMolecularMolecularMolecular formulaformulaformulaformula andandandand molecularmolecularmolecularmolecular weightweightweightweight andandandand chemicalchemicalchemicalchemical structurestructurestructurestructure formulaformulaformulaformula

Molecular formula: C25H26N9NaO8S2

Molecular weight: 667.66

Structure formula:

III.III.III.III. UltraviletUltraviletUltraviletUltravilet absorptionabsorptionabsorptionabsorption spectrumspectrumspectrumspectrum

Apparatus:Cary 1E violet-visible spectrophotometer,America

Conditions:With 10g/ml and 12g/ml of test solution concentration respectively and 190-600nm

of scanning width.

1.1.1.1. UltravioletUltravioletUltravioletUltraviolet absorptionabsorptionabsorptionabsorption spectrumspectrumspectrumspectrum

a.Ultraviolet absorption spectrum of RS

b.Ultraviolet absorption spectrum of test sample

2.Ultraviolet2.Ultraviolet2.Ultraviolet2.Ultraviolet absorptionabsorptionabsorptionabsorption spectrumspectrumspectrumspectrum datadatadatadata ofofofof referencereferencereferencereference standardstandardstandardstandard

For methanol solution,1=200, 2=229,3=269

3.3.3.3. UltravioletUltravioletUltravioletUltraviolet absorptionabsorptionabsorptionabsorption spectrumspectrumspectrumspectrum datadatadatadata ofofofof testtesttesttest samplesamplesamplesample



For methanol solution,1=200, 2=229,3=269

4.Conclusion:4.Conclusion:4.Conclusion:4.Conclusion: There exists -* and n-* band absorption in UV,which shows that the

compound contains the structure replacing the phenyl and carbonyl.

IV.ElementsIV.ElementsIV.ElementsIV.Elements analysisanalysisanalysisanalysis

Apparatus model:EA 1112E element analyzer,Flash company,Italia.

Test condition:Left oven temperature of reactor:950 C;Right oven temperature of reactor:

22C;Gas:High purity He and O2;Chromatographic column temperature: 75C.

Test data list of test sample:

Theoretical value Test value

Test I Test II Test III

C%H%N%S%

44.973.9318.889.61

43.374.2718.769.23

43.414.3318.729.41

43.394.3018.749.32

V.V.V.V. InfraredInfraredInfraredInfrared absorptionabsorptionabsorptionabsorption spectrum(IR)spectrum(IR)spectrum(IR)spectrum(IR)

Apparatus model:FTS-135 Fourier transform infrared spectrometer,BIO-RAD company,America.

Test condition:Adopt KBr pellet method

1.Infrared1.Infrared1.Infrared1.Infrared spectrogramspectrogramspectrogramspectrogram

a. Infrared spectrogram of RS

b.Infrared spectrum of test sample

2.2.2.2. InfraredInfraredInfraredInfrared spectrumspectrumspectrumspectrum testtesttesttest datadatadatadata tabletabletabletable ofofofof testtesttesttest samplesamplesamplesample



Note:strength: vs: extremely strong,s:strong,m:medium,w:weak



3.3.3.3. IRIRIRIR SpectrumSpectrumSpectrumSpectrum analysisanalysisanalysisanalysis

a.Wide and strong absorption centered at ~3400cm-1 is originated from OH stretching

vibration;Strong absorption entrapped in low wave number side of 1610 cm-1 and 1514cm-1 is

originated from benzene ring vibration(Vring);The OH changing angular vibration at 1365cm-1 and

C-O stretching vibration at 1174cm-1 is originated from phenol;The absorption at 841cm-1 is

originated from outside changing angular viabration of C-H side.All these data show that there

exists structure in the molecule.

b.Wide and strong absorption at 3287 cm-1 is originated from NH stretching

vibration(vNH);1680cm-1 is originated from amid I band;Absorption entrapped in 1514cm-1 is

originated from absorption on amid II band;Amid III band is at 1282cm-1 ;806cm-1 and 753cm-1 is

originated separately from amid V band and IV band;576cm-1 is originated from amid VI

band.Appearance of all these bands is connective with the amid structure .

c.Stretching virbration with high wave number C=O(vC=O) appearing at 1762 cm-1 is

characterization of structure.

d.1716cm-1 and 1670cm-1 (entrapped in the low wave number side of 1610 cm-1)is separatedly

originated from stretching vibration of two carbonyls in structure ,together considering

that characteristic vibration of CH2-CH2- appears at 753cm-1 and CH stretching vibration

(vas CH2)of CH2 at 2940cm-1 and changing angular vibration of CH2 at 1463cm-1 ,all these prove

that there exists structure in the molecule.

e.Addtionally,absorption band at 1610cm-1 involves N=N stretching vibration,while

1514cm-1 ,1365cm-1 ,1282cm-1 and 1109cm-1 is originated from stretching vibration of

ring,showing that there exists this structure in the molecule.697cm-1 and 618cm-1

can be classified as C-S stretching vibration,which is consistent with CH2-S- and

structure .

f.Absorption about at 1540cm-1 entrapped in the high frequency side of 1514cm-1 js



antisymmetry stretching vibration of carboxylic radical COO-(vas COO-).Together with the

symmetry stretching vibration(vas COO-)at 1392cm-1 ,all these prove there exists carboxylic

radical conjugating with C=C in the molecule.

g.IR spectrum of the test sample is consistent with that of the reference standard.

VI.VI.VI.VI.1111HHHH NMRNMRNMRNMR

Apparatus: Unity-400 nuclear magnetic resonance spectrometer,Varian,America.

Solvent:Deuterated dimethyl sulfoxide DMSO-d6 , standard reference

displacement:TMS=0.00;heavy water(D2O),reference displacementD20=4.7

i.i.i.i. One-dimensionalOne-dimensionalOne-dimensionalOne-dimensional 1111HHHH NMRNMRNMRNMR testtesttesttest

1. One-dimensional 1H NMR test

a. 1H NMR spectrum of reference standard

b. Heavy water exchange 1H NMR spectrogram of reference standard

c. 1H NMR spectrum of test sample

d. Heavy water exchange 1H NMR spectrogram of test sample

2. 1H NMR test data table of test sampleSerial no. ofpeak

Chemical shift

Multiplicity Couplingconstant(Hz)

Protonnumber

Correspondingproton

12345678910111213

9.939.729.307.206.745.565.514.874.414.213.913.71-3.321.08

bsddddddddddsmt

6.88.07.88.06.8,4.47.24.412.012.0

7.2

1112211111383

-OH>NH>CH=CH-=CH->CH->CH->CH--CH2--CH2--CH3-CH2--CH3

ii.ii.ii.ii. Two-dimensionalTwo-dimensionalTwo-dimensionalTwo-dimensional 1111H-H-H-H-1111HHHH correlatedcorrelatedcorrelatedcorrelated NMRNMRNMRNMR test(test(test(test(1111H-H-H-H-1111HHHH COSY)COSY)COSY)COSY)

1.Two-dimensional 1H-1H correlated NMR spectrogram of reference standard

2. Two-dimensional 1H-1H correlated NMR spectrogram of test sample

3. Two-dimensional 1H-1H correlated NMR test data table of test sample

Serial no. Chemical shift Correlated peak shift

Crossing peak intensity



12345678910

9.729.307.206.745.565.514.414.213.371.08

5.515.566.747.209.309.724.214.411.083.37

mwsswmssss

iii.iii.iii.iii. ProtonProtonProtonProton NMRNMRNMRNMR spectrumspectrumspectrumspectrum analysisanalysisanalysisanalysis ofofofof testtesttesttest samplesamplesamplesample

1. For 26 protons observed in the 1H NMR spectrum of DMSO-d6 solvent of this product,three

groups of peak at low field will disappear in heavy water,the other peak will become wider

apparently and part of them have shifted. The amount and type of hydrogen atom of the

test sample is consistent with that of the chemical structural formula of Cefoperazone sodium.

2. 9.72 and9.30 at low field is separately weakly coupled with5.51 and5.53 in COSY

spectrum,with the coupling constant separately being 6.8Hz and 8.0Hz,but they will disappear

in the heavy water spectrum.Obviously,the two above-mentioned peaks belong to NH3,which

shows that there exist NHCH structure in this compound.

3. Both7.20 and6.74 belong to d desintegration,and JHH equals to 8Hz.Both peaks correspond

to 2 protons,which produce strong coupling information in COSY spectrum and belong to para

substitution benzene proton contribution.

4. 1.08 at high field corresponds to 3 proton contribution,and belongs to triple

disintegration,which produces coupling information with 3.37 in COSY spectrum and

belongs to CH3 proton contribution.All these show that there exist CH2CH3 structure. 3.91 is

a single peak and its signal strength corresponds to 3 proton resonance,which falls within

lowest field and belongs to NCH3 proton contribution.

5. Two groups of peaks of 4.41 and4.21 constitute d disintegration with same coupling

constant,which produce strong coupling information in COSY spectrum.The two groups of

peaks constitute quadri peaks which show high inside and low outside,and is represented as

typical AB self-rotating system.

6. 1H NMR spectrum of the test sample is consistent with that of the reference standard.

VII.VII.VII.VII.13131313CCCC NMRNMRNMRNMR

Apparatus: Unity-400 nuclear magnetic resonance spectrometer



Solvent:Deuterated water(D2O), reference standard:DMSO=39.5

i.i.i.i. One-dimensionalOne-dimensionalOne-dimensionalOne-dimensional 13131313CCCC NMRNMRNMRNMR testtesttesttest

1. One-dimensional 13C NMR test

a.13C NNE NMR spectrum of reference standard

b. 13C DEPT NMR spectrum of reference standard

c. 13C NNE NMR spectrum of test sample

d. 13C DEPT NMR spectrum of test sample

2.13C NMR test data table of test sample

Serial no. ofpeak

Chemical shift Multiplicity Amount of C Correponding type of C

12345678910111213141516171819202122

171.28166.53163.25159.03155.78155.67153.04151.94129.81127.85126.68117.62115.0457.8457.0356.6042.3439.7035.4733.1425.7010.44

sssssssssdssddddtttqtq

1111111112112111211111

>C=O=CC=O>C=O>C=O>C=O>C=O=C

Documents

CTD Open Cefoperazone