EP 2.6.14 Bacterial Endotoxins

EUROPEAN PHARMACOPOEIA 6.0Imipenem IMPURITIES Specified impurities :A,B,C,E,F.Other detectable impurities:D.Relatedsubstances test AA.3-[(2-chloroethyl)amino]propyl dihydrogen phosphate,B.bis[3-[(2-chloroethyl)amino]propyl]dihydrogen diphosphate,C.R =Cl:2-chloroethanamine,D.R =OH:2-aminoethanol.Related substances test BE.3-chloro-N -(2-chloroethyl)propan-1-amine,F.(RS )-2-chloro-3-(2-chloroethyl)-1,3,2-oxazaphosphinane 2-oxide.01/2008:1226corrected 6.0IMIPENEM Imipenemum C 12H 17N 3O 4S,H 2O M r 317.4DEFINITION (5R ,6S )-6-[(R )-1-Hydroxyethyl]-3-[[2-[(iminomethyl)amino]-ethyl]sulphanyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid monohydrate.Semi-synthetic product derived from a fermentation product.Content :98.0per cent to 102.0per cent (anhydrous substance).CHARACTERS Appearance :white or almost white or pale yellow powder.Solubility :sparingly soluble in water,slightly soluble in methanol.IDENTIFICATIONInfrared absorption spectrophotometry (2.2.24).Comparison :imipenem CRS .TESTSAppearance of solution .The solution is not more opalescentthan reference suspension II (2.2.1)and not more intenselycoloured than intensity 6of the range of the reference solutions of the most appropriate colour (2.2.2,Method II ).Dissolve 0.500g in phosphate buffer solution pH 7.0R3anddilute to 50ml with the same solution.pH (2.2.3):4.5to 7.0.Dissolve 0.500g in carbon dioxide-free water R and diluteto 100.0ml with the same solvent.Specificoptical rotation (2.2.7):+84to +89(anhydroussubstance),measured at 25°C.Dissolve 0.125g in phosphate buffer solution pH 7.0R3anddilute to 25.0ml with the same solution.Relatedsubstances .Liquid chromatography(2.2.29).Keep the solutions in an ice-bath and use within 8h of preparation.Solvent mixture .Mix 0.7volumes of acetonitrile R and99.3volumes of a 0.135g/l solution of dipotassiumhydrogen phosphate R adjusted to pH 6.8with dilutephosphoric acid R .Test solution .Dissolve 40.0mgof the substance to be examined in the solvent mixture and dilute to 100.0ml withthe solvent mixture.Reference solution (a).Dissolve 40.0mg of imipenem CRSin the solvent mixture and dilute to 100.0ml with the solvent mixture.Referencesolution (b).Dilute 1.0ml of the test solution to100.0ml with the solvent mixture.Reference solution (c).Heat 20ml of the test solution,previously adjusted to pH 10with sodium hydroxide solutionR ,at 80°C for 5min (in situ preparation ofimpurity A).Column :—size :l =0.25m,Ø=4.6mm;—stationary phase :octadecylsilyl silica gel forchromatography R (5µm).Mobile phase :mix 0.7volumes of acetonitrileR and99.3volumesof a 8.7g/l solution of dipotassium hydrogenphosphate R adjusted to pH 7.3with dilute phosphoric acid R .Flow rate :1.0ml/min.Detection :spectrophotometer at 254nm.Injection:20µl of the test solution and reference solutions (b)and (c).Run time :twice the retention time of imipenem.Relative retention with reference to imipenem (retention time =about 9min):impurity A =about 0.8.System suitability :reference solution (c):—resolution :minimum 3.5between the peaks due to impurity A and imipenem.Limits :—impurityA :not more than the area ofthe principal peak in the chromatogram obtained with reference solution (b)(1per cent);General Notices (1)apply to all monographs and other texts 2125Imipramine hydrochloride EUROPEAN PHARMACOPOEIA6.0—any other impurity :for each impurity,not more than 0.3times the area of the principal peak in the chromatogram obtained with reference solution (b)(0.3per cent);—sum of impurities other than A :not more than the areaof the principal peak in the chromatogram obtained with reference solution (b)(1per cent);—disregard limit :0.1times the area of the principal peak in the chromatogram obtained with reference solution (b)(0.1per cent).Water (2.5.12):5.0per cent to 8.0per cent,determinedon e an iodosulphurous reagent containing imidazole instead of pyridine and a clean container for each determination.Sulphated ash (2.4.14):maximum 0.2per cent,determinedon 1.0g.Bacterial endotoxins (2.6.14):less than 0.17IU/mg,if intended for usein the manufacture of parenteral dosage forms without a further appropriate procedure for removal of bacterial endotoxins.ASSAY Liquid chromatography (2.2.29)as described in the test for related substances with the following modifications.Injection :test solution and reference solution (a).System suitability :reference solution (a):—repeatability :maximum relative standard deviation of 1.0per cent after 6injections.STORAGE In an airtight container,at a temperature of 2°C to8°C.If the substance is sterile,store in a sterile,airtight,tamper-proof container.IMPURITIES Specified impurities :A.A.(5R ,6S )-3-[(2-aminoethyl)sulphanyl]-6-[(R )-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid (thienamycin).01/2008:0029corrected 6.0IMIPRAMINE HYDROCHLORIDE Imipraminihydrochloridum C 19H 25ClN 2M r 316.9[113-52-0]DEFINITIONImipraminehydrochloride contains not less than 98.5per centand not more than the equivalent of 101.0per cent of 3-(10,11-dihydro-5H -dibenzo[b,f ]azepin-5-yl)-N ,N -dimethylpropan-1-amine hydrochloride,calculated withreference to the dried substance.CHARACTERSAwhite or slightly yellow,crystalline powder,freely soluble in water and in alcohol.IDENTIFICATIONFirst identification:A,C,F.Second identification:A,B,D,E,F.A.Melting point (2.2.14):170°C to 174°C.B.Dissolve20mg in 0.01M hydrochloric acid and dilute to 100.0ml with the same acid.Dilute 1.0ml of the solutionto 10.0mlwith 0.01M hydrochloric acid .Examinedbetween 230nm and 350nm,the solution shows a singleabsorption maximum (2.2.25),at 251nm,and a shoulder at 270nm.The specific absorbance at the maximum isabout 260.C.Examine by infrared absorption spectrophotometry (2.2.24),comparing with the spectrum obtained with imipramine hydrochloride CRS .Examine the substances prepared as discs.D.Dissolveabout 5mg in 2ml of nitric acid R .An intense blue colour develops.E.Dissolve about 50mg in 3mlof water R and add 0.05mlof a 25g/l solution of quinhydrone R in methanol R .Nored colour develops within 15min.F.About 20mg gives reaction (a)of chlorides (2.3.1).TESTSSolution S .To 3.0g add 20ml of carbon dioxide-freewater R ,dissolve rapidly by shaking and triturating with aglass rod and dilute to 30ml with the same solvent.Appearance of solution .SolutionS is clear (2.2.1).Immediately afterpreparation,dilute solution S with anequal volume of water R .This solution is not more intensely coloured than reference solution BY 6(2.2.2,Method II ).pH (2.2.3).The pH of solution S,measured immediately after preparation,is 3.6to 5.0.Related substances .Examine by thin-layer chromatography(2.2.27),using a TLC silica gel G plate R .Testsolution.Dissolve 0.25g of the substance to be examined in methanol R and dilute to 10ml with the samesolvent.Prepare immediately before use.Referencesolution (a).Dilute 1ml of the test solution to10ml with methanol R .Dilute 1ml of this solution to 50mlwith methanol R .Reference solution (b).Dissolve 5mgof iminodibenzyl Rin methanol R and dilute to 100ml with the same solvent.Prepare immediately before use.Apply to the plate10µl of each solution.Develop over apath of 12cm using a mixture of 5volumes of hydrochloricacid R ,5volumes of water R ,35volumes of glacial acetic acid R and 55volumes of ethyl acetate R .Allow the plateto dry in air for5min and spray with a 5g/l solutionof potassium dichromate R in a mixture of 1volume ofsulphuric acid R and 4volumes of water R .Examine the plate immediately.The chromatogram obtained with the test solution shows a blue principal spot.In the chromatogram obtained with the test solution:any spot corresponding 2126See the information section on general monographs (cover pages)。

EUROPEAN PHARMACOPOEIA 8.0FramycetinsulfateI.[(RS )-[(1SR )-2-methyl-1-(1-oxopropoxy)propoxy]-(4-phenylbutyl)phosphoryl]aceticacid,K.(2S ,4S )-4-cyclohexyl-1-(2,2-dimethyl-1-oxopropyl)pyrroli-dine-2-carboxylicacid,N.(2S ,4S )-4-cyclohexyl-1-[[(2S ,4S )-4-cyclohexyl-1-[[(R )-[(1S )-2-methyl-1-(1-oxopropoxy)propoxy](4-phenylbutyl)phosphoryl]acetyl]pyrrolidin-2-yl]-carbonyl]pyrrolidine-2-carboxylic acid.01/2008:0180FRAMYCETIN SULFATEFramycetinisulfasC 23H 46N 6O 13,x H 2SO 4M r 615(base)[4146-30-9]DEFINITIONSulfate of 2-deoxy-4-O -(2,6-diamino-2,6-dideoxy-α-D -glucopyranosyl)-5-O -[3-O -(2,6-diamino-2,6-dideoxy-β-L -idopyranosyl)-β-D -ribofuranosyl]-D -streptamine (neomycin B).Substance produced by the growth of selected strains of Streptomyces fradiae or Streptomyces decaris or obtained byany other means.Content :minimum of 630IU/mg (dried substance).CHARACTERSAppearance :white or yellowish-white powder,hygroscopic.Solubility :freely soluble in water,very slightly soluble inalcohol,practically insoluble in acetone.IDENTIFICATION A.Examine the chromatograms obtained in the test forrelated substances.Results :–the retention time of the principal peak in the chromatogram obtained with the test solution isapproximately the same as that of the principal peak in the chromatogram obtained with reference solution (a),–it complies with the limit given for impurity C.B.It gives reaction (a)of sulfates (2.3.1).TESTSpH (2.2.3):6.0to 7.0.Dissolve 0.1g in carbon dioxide-free water R and dilute to 10mL with the same solvent.Specific optical rotation (2.2.7):+52.5to +55.5(dried substance).Dissolve 1.00g in water R and dilute to 10.0mL with the samesolvent Related substances .Liquid chromatography (2.2.29).Test solution .Dissolve 25.0mg of the substance to beexamined in the mobile phase and dilute to 50.0mL with the mobile phase.Reference solution (a).Dissolve the contents of a vial offramycetin sulfate CRS in the mobile phase and dilute with the mobile phase to obtain a solution containing 0.5mg/mL.Reference solution (b).Dilute 3.0mL of reference solution (a)to 100.0mL with the mobile phase.Reference solution (c).Dilute 1.0mL of reference solution (a)to 100.0mL with the mobile phase.Reference solution (d).Dissolve the contents of a vial of neamine CRS (corresponding to 0.5mg)in the mobile phase and dilute to 100.0mL with the mobile phase.Reference solution (e).Dissolve 10mg of neomycin sulfate CRS in the mobile phase and dilute to 100.0mL with the mobile phase.Column :–size :l =0.25m,Ø=4.6mm,–stationary phase :base-deactivated octadecylsilyl silica gel for chromatography R (5μm),–temperature :25°C.Mobile phase :mix 20.0mL of trifluoroacetic acid R ,6.0mL of carbonate-free sodium hydroxide solution R and 500mL of water R ,allow to equilibrate,dilute to 1000mL with water R and degas.Flow rate :0.7mL/min.Post-column solution :carbonate-free sodium hydroxidesolution R diluted 1in 25previously degassed,which is added pulse-less to the column effluent using a 375μL polymeric mixing coil.Flow rate :0.5mL/min.Detection :pulsed amperometric detector with a gold workingelectrode,a silver-silver chloride reference electrode and a stainless steel auxiliary electrode which is the cell body,held atrespectively 0.00V detection,+0.80V oxidation and −0.60Vreduction potentials,with pulse durations according to theinstrument used.Injection :10μL.Run time :1.5times the retention time of neomycin B.Relative retention with reference to neomycin B (retention time =about 10min):impurity A =about 0.65;impurity C =about 0.9;impurity G =about 1.1.System suitability :–resolution :minimum 2.0between the peaks due to impurity C and to neomycin B in the chromatogram obtained with reference solution (e);if necessary,adjust the volume of the carbonate-free sodium hydroxide solutionin the mobile phase,–signal-to-noise ratio :minimum 10for the principal peak in the chromatogram obtained with reference solution (c).General Notices (1)apply to all monographs and other texts2305Fructose EUROPEAN PHARMACOPOEIA8.0Limits :–impurity A :not more than the area of the principal peak in the chromatogram obtained with reference solution (d)and taking into account the declared content of neamine CRS (1.0per cent),–impurity C :not more than the area of the principal peak in the chromatogram obtained with reference solution (b)(3.0per cent),–total of other impurities :not more than the area of the principal peak in the chromatogram obtained with reference solution (b)(3.0per cent),–disregard limit :area of the principal peak in thechromatogram obtained with reference solution (c)(1.0per cent).Sulfate :27.0per cent to 31.0per cent (dried substance).Dissolve 0.250g in 100mL of water R and adjust the solution to pH 11using concentrated ammonia R .Add 10.0mLof 0.1M barium chloride and about 0.5mg of phthaleinpurple R .Titrate with 0.1M sodium edetate adding 50mL ofalcohol R when the colour of the solution begins to change andcontinuing the titration until the violet-blue colour disappears.1mL of 0.1M barium chloride is equivalent to 9.606mg of SO 4.Loss on drying (2.2.32):maximum 8.0per cent,determined on 1.000g by drying at 60°C over diphosphorus pentoxide R ata pressure not exceeding 0.7kPa for 3h.Sulfated ash (2.4.14):maximum 1.0per cent,determined on 1.0g.Sterility (2.6.1).If intended for introduction into bodycavities without a further appropriate sterilisation procedure,it complies with the test for sterility.Bacterial endotoxins (2.6.14,Method D ):less than 1.3IU/mgif intended for introduction into body cavities without afurther appropriate procedure for the removal of bacterial endotoxins.ASSAYCarry out the microbiological assay of antibiotics (2.7.2).Use framycetin sulfate CRS as the reference substance.STORAGEIn an airtight container,protected from light.If the substance is intended for introduction into body cavities,store in a sterile,tamper-proof container.IMPURITIESA.R1=H,R2=NH 2:2-deoxy-4-O -(2,6-diamino-2,6-dideoxy-α-D -glucopyranosyl)-D -streptamine (neamine or neomycin A-LP),B.R1=CO-CH 3,R2=NH 2:3-N -acetyl-2-deoxy-4-O -(2,6-diamino-2,6-dideoxy-α-D -glucopyranosyl)-D -streptamine (3-acetylneamine),D.R1=H,R2=OH:4-O -(2-amino-2-deoxy-α-D -glucopyranosyl)-2-deoxy-D -streptamine (paromamine or neomycinD), C.R1=CH 2-NH 2,R2=R3=H,R4=NH 2:2-deoxy-4-O -(2,6-diamino-2,6-dideoxy-α-D -glucopyranosyl)-5-O -[3-O -(2,6-diamino-2,6-dideoxy-α-D -glucopyranosyl)-β-D -ribofuranosyl]-D -streptamine (neomycin C),E.R1=R3=H,R2=CH 2-NH 2,R4=OH:4-O -(2-amino-2-deoxy-α-D -glucopyranosyl)-2-deoxy-5-O -[3-O -(2,6-diamino-2,6-dideoxy-β-L -idopyranosyl)-β-D -ribofuranosyl]-D -streptamine (paromomycin I orneomycin E),F.R1=CH 2-NH 2,R2=R3=H,R4=OH:4-O -(2-amino-2-deoxy-α-D -glucopyranosyl)-2-deoxy-5-O -[3-O -(2,6-diamino-2,6-dideoxy-α-D -glucopyranosyl)-β-D -ribofuranosyl]-D -streptamine (paromomycin II or neomycin F),G.R1=H,R2=CH 2-NH 2,R3=CO-CH 3,R4=NH 2:3-N -acetyl-2-deoxy-4-O -(2,6-diamino-2,6-dideoxy-α-D -glucopyranosyl)-5-O -[3-O -(2,6-diamino-2,6-dideoxy-β-L -idopyranosyl)-β-D -ribofuranosyl]-D -streptamine(neomycin B-LP).01/2008:0188corrected 6.0FRUCTOSEFructosum C 6H 12O 6M r 180.2[57-48-7]DEFINITIOND -arabino -Hex-2-ulopyranose.The substance described in this monograph is not necessarily suitable for parenteral administration.CHARACTERSAppearance :white or almost white,crystalline powder.It has a very sweet taste.Solubility :very soluble in water,soluble in ethanol (96per cent).IDENTIFICATIONA.Thin-layer chromatography (2.2.27).Solvent mixture :water R ,methanol R (2:3V/V ).Test solution .Dissolve 10mg of the substance to beexamined in the solvent mixture and dilute to 20mL with the solvent mixture.Reference solution (a).Dissolve 10mg of fructose CRS in the solvent mixture and dilute to 20mL with the solvent mixture.Reference solution (b).Dissolve 10mg each of fructose CRS ,glucose CRS ,lactose CRS and sucrose CRS in the solvent mixture and dilute to 20mL with the solvent mixture.Plate :TLC silica gel G plate R .2306See the information section on general monographs (cover pages)。

EU,USP及ChP对干热灭菌/除热原条件对照表附件1 -1EP6.0 5.1.1. METHODS OF PREPARATIONOF STERILE PRODUCTS)Dry heat sterilisation. For this method of terminal sterilisation the reference conditions are a minimumof 160 °C for at least 2 h. Other combinations of timeand temperature may be used provided that it has beensatisfactorily demonstrated that the process chosen deliversan adequate and reproducible level of lethality whenoperated routinely within the established tolerances. Theprocedures and precautions employed are such as to give anSAL of 10− 6 or better.Dry heat sterilisation is carried out in an oven equipped withforced air circulation or other equipment specially designedfor the purpose. The steriliser is loaded in such a way thata uniform temperature is achieved throughout the load.Knowledge of the temperature within the steriliser duringthe sterilisation procedure is usually obtained by means oftemperature-sensing elements inserted into representativecontainers together with additional elements at thepreviously established coolest part of the loaded steriliser.The temperature throughout each cycle is suitably recorded.Where a biological assessment is carried out, this is obtainedusing a suitable biological indicator (5.1.2).Dry heat at temperatures greater than 220 °C is frequentlyused for sterilisation and depyrogenation of glassware. Inthis case demonstration of a 3-log reduction in heat resistantendotoxin can be used as a replacement for biologicalindicators (5.1.2)附件1-2 2.6.14. BACTERIAL ENDOTOXINSThe test for bacterial endotoxins is used to detect orquantify endotoxins of gram-negative bacterial originusing amoebocyte lysate from horseshoe crab (Limuluspolyphemus or Tachypleus tridentatus). There are3 techniques for this test : the gel-clot technique, whichis based on gel formation ; the turbidimetric technique,based on the development of turbidity after cleavage of anendogenous substrate ; and the chromogenic technique,based on the development of colour after cleavage of asynthetic peptide-chromogen complex.The following 6 methods are described in the presentchapter :Method A. Gel-clot method: limit testMethod B. Gel-clot method: semi-quantitative testMethod C. Turbidimetric kinetic method182 See the information section on general monographs (cover pages) EUROPEAN PHARMACOPOEIA 6.0 2.6.14. Bacterial endotoxinsMethod D. Chromogenic kinetic methodMethod E. Chromogenic end-point methodMethod F. Turbidimetric end-point methodProceed by any of the 6 methods for the test. In the eventof doubt or dispute, the final decision is made based uponmethod A unless otherwise indicated in the monograph.The test is carried out in a manner that avoids endotoxincontamination.ApparatusDepyrogenate all glassware and other heat-stable apparatusin a hot-air oven using a validated process. A commonly usedminimum time and temperature is 30 minutes at 250 °C. Ifemploying plastic apparatus, such as microtitre plates andpipette tips for automatic pipetters, use apparatus shownto be free of detectable endotoxin and of interfering effectsfor the test.NOTE: In this chapter, the term ‘tube’ includes all types ofreceptacles, for example microtitre plate wells.indicators (5.1.2).附件2-11211STERILIZATION AND STERILITY ASSURANCE OF COMPENDIAL ARTICLESThis informational chapter provides a general description of the concepts and principles involved in the quality control of articles that must be sterile. Any modifications of or variations in sterility test procedures from those described under Sterility Tests 71should be validated in the context of the entire sterility assurance program and are not intended to be methods alternative to those described in that chapter.Within the strictest definition of sterility, a specimen would be deemed sterile only when there is complete absence of viable microorganisms from it. However, this absolute definition cannot currently be applied to an entire lot of finished compendial articles because of limitations in testing. Absolute sterility cannot be practically demonstrated without complete destruction of every finished article. The sterility of a lot purported to be sterile is therefore definedin probabilistic terms, where the likelihood of a contaminated unit or article is acceptably remote. Such a state of sterility assurance can be established only through the use of adequate sterilization cycles and subsequent asepticprocessing, if any, under appropriate current good manufacturing practice, and not by reliance solely on sterility testing. The basic principles for validation and certification of a sterilizing process are enumerated as follows:1. Establish that the process equipment has capability of operating within therequired parameters.2. Demonstrate that the critical control equipment and instrumentation arecapable of operating within the prescribed parameters for the processequipment.3. Perform replicate cycles representing the required operational range of theequipment and employing actual or simulated product. Demonstrate that the processes have been carried out within the prescribed protocol limits andfinally that the probability of microbial survival in the replicate processescompleted is not greater than the prescribed limits.4. Monitor the validated process during routine operation. Periodically as needed,requalify and recertify the equipment.5. Complete the protocols, and document steps (1) through (4) above.METHODS OF STERILIZATIONIn this informational chapter, five methods of terminal sterilization, including removal of microorganisms by filtration and guidelines for aseptic processing, are described. Modern technological developments, however, have led to the use of additional procedures. These include blow-molding (at hightemperatures), forms of moist heat other than saturated steam and UVirradiation, as well as on-line continuous filling in aseptic processing. Thechoice of the appropriate process for a given dosage form or componentrequires a high level of knowledge of sterilization techniques and information concerning any effects of the process on the material being sterilized.1Dry-Heat SterilizationThe process of thermal sterilization of Pharmacopeial articles by dry heat is usually carried out by a batch process in an oven designed expressly for that purpose. A modern oven is supplied with heated, filtered air, distributeduniformly throughout the chamber by convection or radiation and employing a blower system with devices for sensing, monitoring, and controlling the critical parameters. The validation of a dry-heat sterilization facility is carried out in a manner similar to that for a steam sterilizer described earlier. Where the unit is employed for sterilizing components such as containers intended forintravenous solutions, care should be taken to avoid accumulation ofparticulate matter in the chamber. A typical acceptable range in temperature in the empty chamber is ±15when the unit is operating at not less than 250.In addition to the batch process described above, a continuous process is frequently employed to sterilize and depyrogenate glassware as part of an integrated continuous aseptic filling and sealing system. Heat distribution may be by convection or by direct transfer of heat from an open flame. The continuous system usually requires a much higher temperature than cited above for the batch process because of a much shorter dwell time. However, the total temperature input during the passage of the product should be equivalent to that achieved during the chamber process. The continuous process also usually necessitates a rapid cooling stage prior to the aseptic filling operation. In the qualification and validation program, in view of the short dwell time, parameters for uniformity of the temperature, and particularly the dwell time, should be established.A microbial survival probability of 10–12 is considered achievable forheat-stable articles or components. An example of a biological indicator for validating and monitoring dry-heat sterilization is a preparation of Bacillus subtilis spores. Since dry heat is frequently employed to render glassware or containers free from pyrogens as well as viable microbes, a pyrogen challenge, where necessary, should be an integral part of the validation program, e.g., by inoculating one or more of the articles to be treated with 1000 or more USP Units of bacterial endotoxin. The test with Limulus lysate could be used to demonstrate that the endotoxic substance has been inactivated to not more than 1/1000 of the original amount (3 log cycle reduction). For the test to be valid, both the original amount and, after acceptable inactivation, the remaining amount of endotoxin should be measured. For additional information on the endotoxin assay, see Bacterial Endotoxins Test 85.附件2-2 85BACTERIAL ENDOTOXINS TESTAPPARATUS AND GLASSWAREDepyrogenate all glassware and other heat-stable materials in a hot-air oven using a validated process.2Commonly used minimum time and temperature settings are 30 minutes at 250. If employing plastic apparatus, such as microplates and pipet tips for automatic pipetters, use only that which has been shown to be free of detectable endotoxin and not to interfere with the test. [note—In this chapter, the term “tube” includes any other receptacle such as a micro-titer well.]。

EUROPEAN PHARMACOPOEIA 7.0Xylitol—mobile phase B :methanol R ,acetonitrile R (30:70V/V );Time (min)Mobile phase A (per cent V/V )Mobile phase B (per cent V/V )0-1589→2811→7215-212872Flow rate :1.0mL/min.Detection :spectrophotometer at 230nm.Equilibration :with a mixture of 28volumes of mobile phase A and 72volumes of mobile phase B for at least 30min.Injection :20μL.Identification of impurities :use the chromatogram supplied with xylazine impurity mixture CRS and the chromatogram obtained with reference solution (b)to identify the peaks due to impurities B and D;use the chromatogram obtained with reference solution (a)to identify the peaks due to impurities A,C and E.Relative retention with reference to xylazine (retention time =about 7.5min):impurity D =about 0.5;impurity A =about 0.8;impurity B =about 1.3;impurity E =about 1.6;impurity C =about 2.2.System suitability :reference solution (a):—resolution :minimum 4.0between the peaks due to impurity A and xylazine.Limits :—impurities B,D :for each impurity,not more than twice the area of the peak due to xylazine in the chromatogram obtained with reference solution (a)(0.2per cent);—impurities C,E :for each impurity,not more than twice the area of the corresponding peak in the chromatogram obtained with reference solution (a)(0.2per cent);—unspecified impurities :for each impurity,not more than twice the area of the peak due to xylazine in thechromatogram obtained with reference solution (a)(0.2per cent);—total of impurities other than B,C,D and E :not more than twice the area of the peak due to xylazine in thechromatogram obtained with reference solution (a)(0.2per cent);—disregard limit :0.5times the area of the peak due to xylazine in the chromatogram obtained with reference solution (a)(0.05per cent);disregard any peak due to the blank.Heavy metals (2.4.8):maximum 10ppm.12mL of solution S complies with test A.Prepare the reference solution using 10mL of lead standard solution (1ppm Pb)R .Loss on drying (2.2.32):maximum 0.5per cent,determined on 1.000g by drying in an oven at 105°C for 2h.Sulfated ash (2.4.14):maximum 0.1per cent,determined on 1.0g.ASSAY Dissolve 0.200g in 25mL of ethanol (96per cent)R .Add 25mL of water R .Titrate with 0.1M sodium hydroxide ,determiningthe end-point potentiometrically (2.2.20).1mL of 0.1M sodium hydroxide is equivalent to 25.68mgof C 12H 17ClN 2S.STORAGEIn an airtight container,protected from light.IMPURITIESSpecified impurities:A,B,C,D,E.A.2,6-dimethylaniline(2,6-xylidine),B.N ,N ′-bis(2,6-dimethylphenyl)thiourea,C.2,6-dimethylphenylisothiocyanate,D.N -(2,6-dimethylphenyl)-N ′-(3-hydroxypropyl)thiourea,E.methyl (2,6-dimethylphenyl)carbamodithioate.01/2009:1381XYLITOLXylitolumC 5H 12O 5M r 152.1[87-99-0]DEFINITION Meso -xylitol.Content :98.0per cent to 102.0per cent (anhydrous substance).CHARACTERS Appearance :white or almost white,crystalline powder or crystals.Solubility :very soluble in water,sparingly soluble in ethanol (96per cent).IDENTIFICATION First identification:B.Second identification:A,C.A.Melting point (2.2.14):92°C to 96°C.B.Infrared absorption spectrophotometry (2.2.24).Preparation :mulls in liquid paraffin R .Comparison :xylitol CRS .C.Thin-layer chromatography (2.2.27).Test solution .Dissolve 25mg of the substance to be examined in water R and dilute to 5mL with the same solvent.Reference solution (a).Dissolve 25mg of xylitol CRS inwater R and dilute to 5mL with the same solvent.General Notices (1)apply to all monographs and other texts3239Xylitol EUROPEAN PHARMACOPOEIA7.0Reference solution(b).Dissolve25mg of mannitol CRSand25mg of xylitol CRS in water R and dilute to5mL with the same solvent.Plate:TLC silica gel G plate R.Mobile phase:water R,ethyl acetate R,propanol R(10:20:70V/V/V).Application:2μL.Development:over3/4of the plate.Drying:in air.Detection:spray with4-aminobenzoic acid solution R,dry in a current of cold air until the acetone is removed,then heat at100°C for15min;allow to cool,spray with a2g/L solution of sodium periodate R,dry in a current of cold air, then heat at100°C for15min.System suitability:reference solution(b):—the chromatogram shows2clearly separated spots.Results:the principal spot in the chromatogram obtained with the test solution is similar in position,colour and size to the principal spot in the chromatogram obtained withreference solution(a).TESTSAppearance of solution.The solution is not more opalescent than reference suspension IV(2.2.1)and not more intensely coloured than reference solution BY7(2.2.2,Method II). Dissolve2.5g in water R and dilute to50.0mL with the same solvent.Conductivity(2.2.38):maximum20μS·cm−1.Dissolve20.0g in carbon dioxide-free water R prepared from distilled water R and dilute to100.0mL with the same solvent. Measure the conductivity of the solution while gently stirring with a magnetic stirrer.Reducing sugars:maximum0.2per cent,calculated as glucose equivalent.Dissolve5.0g in6mL of water R with the aid of gentle heat. Cool and add20mL of cupri-citric solution R and a few glass beads.Heat so that boiling begins after4min and maintain boiling for3min.Cool rapidly and add100mL of a2.4per cent V/V solution of glacial acetic acid R and20.0mL of0.025M iodine.With continuous shaking,add25mL of a mixture of6volumes of hydrochloric acid R and94volumes of water R and,when the precipitate has dissolved,titrate the excess of iodine with0.05M sodium thiosulfate using1mL of starch solution R,added towards the end of the titration,as indicator.Not less than12.8mL of0.05M sodium thiosulfate is required.Related substances.Gas chromatography(2.2.28).Internal standard solution.Dissolve5mg of erythritol R in water R and dilute to25.0mL with the same solvent.Test solution(a).Dissolve5.000g of the substance to be examined in water R and dilute to100.0mL with the same solvent.Test solution(b).Dilute1.0mL of test solution(a)to10.0mL with water R.Reference solution(a).Dissolve5.0mg each of L-arabinitol CRS (impurity A),galactitol CRS(impurity B),mannitol CRS (impurity C)and sorbitol CRS(impurity D)in water R and dilute to20.0mL with the same solvent.Reference solution(b).Dissolve50.0mg of xylitol CRS in water R and dilute to10.0mL with the same solvent.Pipette1.0mL of test solutions(a)and(b)and reference solutions(a)and(b)into4separate100mL round-bottomed flasks.Add1.0mL of the internal standard solution to each of the flasks containing test solution(a)or reference solution(a), and5.0mL of the internal standard solution to each of the flasks containing test solution(b)or reference solution(b). Evaporate each mixture to dryness in a water-bath at60°C with the aid of a rotary evaporator.Dissolve each dry residue in 1mL of anhydrous pyridine R,add1mL of acetic anhydride R to each flask and boil each solution under reflux for1h to complete acetylation.Column:—size:l=30m,Ø=0.25mm;—stationary phase:poly(cyanopropylphenyl)(14)(methyl)-(86)siloxane R(0.25μm).Carrier gas:nitrogen R.Flow rate:1mL/min.Split ratio:1:50to1:100.Temperature:Time(min)Temperature(°C)Column0-11701-6170→2306-30230Injection port250Detector250 Detection:flame-ionisation.Injection:1μL of test solution(a)and reference solution(a) (solutions obtained after derivatisation).Relative retention with reference to xylitol(retentiontime=about15min):internal standard=about0.6; impurity A=about0.9;impurity C=about1.4;impurity B=about1.45;impurity D=about1.5.System suitability:reference solution(a):—resolution:minimum2.0between the peaks due to impurities B and D.Calculate the percentage content of each related substance in the substance to be examined using the following expression:ms=mass of the particular component in1mL ofreference solution(a),in milligrams;mu=mass of the substance to be examined in1mL of test solution(a),in milligrams;Rs=ratio of the area of the peak due to the particular derivatised component to the area of the peakdue to the derivatised internal standard in thechromatogram obtained with reference solution(a);Ru=ratio of the area of the peak due to the particular derivatised component to the area of the peakdue to the derivatised internal standard in thechromatogram obtained with test solution(a).The sum of the percentage contents of the related substances in the chromatogram obtained with test solution(a)is not greater than2.0per cent.Disregard any peak with an area corresponding to a percentage content of0.05per cent or less. Lead(2.4.10):maximum0.5ppm.Dissolve the substance to be examined in150.0mL of the prescribed mixture of solvents.Nickel(2.4.15):maximum1ppm.Dissolve the substance to be examined in150.0mL of the prescribed mixture of solvents.Water(2.5.12):maximum1.0per cent,determined on1.00g. Bacterial endotoxins(2.6.14):less than4IU/g if the concentration is less than100g/L of xylitol and less than2.5IU/g if the concentration is100g/L or more of xylitol,when intended for use in the manufacture of parenteral preparations without a further appropriate procedure for the removal of bacterial endotoxins.3240See the information section on general monographs(cover pages)EUROPEAN PHARMACOPOEIA 7.0XylometazolinehydrochlorideASSAYGas chromatography (2.2.28)as described in the test for related substances with the following modifications.Injection :1μL of test solution (b)and reference solution (b)(solutions obtained after derivatisation).Calculate the percentage content of C 5H 12O 5using the following expression:T =declared percentage content of xylitol CRS ;m t =mass of xylitol CRS in 1mL of reference solution (b),in milligrams;m v =mass of the substance to be examined in 1mL of test solution (b),in milligrams;R t=ratio of the area of the peak due to derivatised xylitol to the area of the peak due to the derivatised internal standard in the chromatogram obtained with reference solution (b);R v=ratio of the area of the peak due to derivatised xylitol to the area of the peak due to the derivatised internal standard in the chromatogram obtained with test solution (b).LABELLINGThe label states:—where applicable,the maximum concentration of bacterial endotoxins;—where applicable,that the substance is suitable for use in themanufacture of parenteral preparations.IMPURITIESA.L-arabinitol,B.meso-galactitol,C.D-mannitol,D.D -glucitol (D -sorbitol).01/2008:1162corrected 7.0XYLOMETAZOLINE HYDROCHLORIDEXylometazolinihydrochloridumC 16H 25ClN 2M r 280.8[1218-35-5]DEFINITION2-[4-(1,1-Dimethylethyl)-2,6-dimethylbenzyl]-4,5-dihydro-1H -imidazole hydrochloride.Content :99.0per cent to 101.0per cent (dried substance).CHARACTERSAppearance :white or almost white,crystalline powder.Solubility :freely soluble in water,in ethanol (96per cent)and in methanol.IDENTIFICATIONFirst identification:A,E.Second identification:B,C,D,E.A.Infrared absorption spectrophotometry (2.2.24).Comparison :xylometazoline hydrochloride CRS .B.Thin-layer chromatography (2.2.27).Test solution .Dissolve 20mg of the substance to be examined in methanol R and dilute to 5mL with the same solvent.Reference solution .Dissolve 20mg of xylometazoline hydrochloride CRS in methanol R and dilute to 5mL with the same solvent.Plate :TLC silica gel G plate R .Mobile phase :concentrated ammonia R ,methanol R (5:100V/V ).Application :5μL.Development :over 2/3of the plate.Drying :in air.Chlorine treatment :at the bottom of a chromatographic tank place a beaker containing a mixture of 1volume of hydrochloric acid R1,1volume of water R and 2volumes of a 15g/L solution of potassium permanganate R .Close the tank and allow to stand for 15min.Place the dried plate in the tank and reclose the tank.Leave the plate in contact with the chlorine vapour for 5min.Withdraw theplate and place it in a current of cold air until the excess ofchlorine is removed and an area of the coating below the points of application does not give a blue colour with a drop of potassium iodide and starch solution R .Detection :spray with potassium iodide and starch solution R .Results :the principal spot in the chromatogram obtained with the test solution is similar in position,colour and size to the principal spot in the chromatogram obtained with the reference solution.C.Dissolve about 0.5mg in 1mL of methanol R .Add 0.5mL of a freshly prepared 50g/L solution of sodium nitroprusside R and 0.5mL of a 20g/L solution of sodium hydroxide R .Allow to stand for 10min and add 1mL of an 80g/L solution of sodium bicarbonate R .A violet colour develops.D.Dissolve 0.2g in 1mL of water R ,add 2.5mL of ethanol (96per cent)R and 2mL of 1M sodium hydroxide .Mixthoroughly and examine in ultraviolet light at 365nm.General Notices (1)apply to all monographs and other texts3241。

EUROPEAN PHARMACOPOEIA 6.0Water,purifieding Table 1927.-2,determine the conductivity limit at the measured pH value in step 6.If the measured conductivity in step 4under stage 2is not greater than the conductivity requirements for the pH determined,the water to be examined meets the requirements of the test for conductivity.If either the measured conductivity is greater than this value or the pH is outside the range of 5.0-7.0,the water to be examined does not meet the requirements of the test for conductivity.In order to ensure the appropriate quality of the water,validated procedures and in-process monitoring of theelectrical conductivity and regular microbial monitoring are applied.Highly purified water is stored in bulk and distributed in conditions designed to prevent growth of micro-organisms and to avoid any other contamination.Table 1927.-2.–Stage 3-pH and conductivity requirements(for atmosphere and temperature equilibrated samples)pH Conductivity(µS·cm −1)5.0 4.75.1 4.15.2 3.65.3 3.35.4 3.05.5 2.85.6 2.65.7 2.55.8 2.45.9 2.46.0 2.46.1 2.46.2 2.56.3 2.46.4 2.36.5 2.26.6 2.16.7 2.66.8 3.16.9 3.87.04.6CHARACTERSAppearance :clear and colourless liquid.TESTSNitrates :maximum 0.2ppm.Place 5ml in a test-tube immersed in iced water,add 0.4ml of a 100g/l solution of potassium chloride R ,0.1ml of diphenylamine solution R and,dropwise with shaking,5ml of nitrogen-free sulphuric acid R .Transfer the tube to a water-bath at 50°C.After 15min,any blue colour in the solution is not more intense than that in a reference solution prepared at the same time in the same manner using a mixture of 4.5ml of nitrate-free water R and 0.5ml of nitrate standard solution (2ppm NO 3)R .Aluminium (2.4.17):maximum 10ppb,if intended for use in the manufacture of dialysis solutions.Prescribed solution .To 400ml of the water to be examined add 10ml of acetate buffer solution pH 6.0R and 100ml of distilled water R .Reference solution .Mix 2ml of aluminium standard solution (2ppm Al)R ,10ml of acetate buffer solution pH 6.0R and 98ml of distilled water R .Blank solution .Mix 10ml of acetate buffer solution pH 6.0R and 100ml of distilled water R .Heavy metals (2.4.8):maximum 0.1ppm.Heat 200ml in a glass evaporating dish on a water-bath until the volume is reduced to 20ml.12ml of the concentrated solution complies with limit test A.Prepare the standard using 10ml of lead standard solution (1ppm Pb)R .Bacterial endotoxins (2.6.14):less than 0.25IU/ml.LABELLING The label states,where applicable,that the substance is suitable for use in the manufacture of dialysis solutions.01/2008:0008WATER,PURIFIED Aqua purificataH 2OM r 18.02[7732-18-5]DEFINITIONWater for the preparation of medicines other than those that are required to be both sterile and apyrogenic,unless otherwise justified and authorised.Purified water in bulkPRODUCTIONPurified water in bulk is prepared by distillation,by ion exchange,by reverse osmosis or by any other suitable method from water that complies with the regulations on water intended for human consumption laid down by the competent authority.During production and subsequent storage,appropriate measures are taken to ensure that the total viable aerobic count is adequately controlled and monitored.Appropriate alert and action limits are set so as to detect adverse trends.Under normal conditions,an appropriate action limit is a total viable aerobic count (2.6.12)of 100micro-organisms per millilitre,determined by membrane filtration,using agar medium S and incubating at 30-35°C for 5days.The size of the sample is to be chosen in relation to the expected result.In addition,the test for total organic carbon (2.2.44)with a limit of 0.5mg/l or alternatively the following test for oxidisable substances is carried out:to 100ml add 10ml of dilute sulphuric acid R and 0.1ml of 0.02M potassium permanganate and boil for 5min;the solution remains faintly pink.Conductivity .Determine the conductivity off-line or in-line under the following conditions.EQUIPMENT Conductivity cell :—electrodes of a suitable material such as stainless steel;General Notices (1)apply to all monographs and other texts3213Water,purified EUROPEAN PHARMACOPOEIA6.0—cell constant:within2per cent of the given value determined using a certified reference solution with aconductivity less than1500µS·cm−1. Conductometer:resolution0.1µS·cm−1on the lowest range. System calibration(conductivity cell and conductometer):—against one or more suitable certified standard solutions;—accuracy:within3per cent of the measured conductivity plus0.1µS·cm−1.Conductometer calibration:by means of precision resistors or equivalent devices,after disconnecting the conductivity cell,for all ranges used for conductivity measurement and cell calibration(with an accuracy within0.1per cent of the stated value,traceable to the official standard).If in-line conductivity cells cannot be dismantled,system calibration may be performed against a calibrated conductivity cell placed close to the cell to be calibrated in the water flow.PROCEDUREMeasure the conductivity without temperature compensation,recording simultaneously the temperature. Temperature-compensated measurement may be performed after suitable validation.The water to be examined meets the requirements if the measured conductivity at the recorded temperature is not greater than the value in Table0008.-1.Table0008.-1.–Temperature and conductivityrequirementsTemperature(°C)Conductivity (µS·cm−1)0 2.410 3.620 4.325 5.130 5.440 6.5507.1608.1709.1759.7809.7909.710010.2For temperatures not listed in Table0008.-1,calculate the maximal permitted conductivity by interpolation between the next lower and next higher data points in the table. Purified water in bulk is stored and distributed in conditions designed to prevent growth of micro-organisms and to avoid any other contamination.CHARACTERSAppearance:clear and colourless liquid.TESTSNitrates:maximum0.2ppm.Place5ml in a test-tube immersed in iced water,add0.4ml of a100g/l solution of potassium chloride R,0.1ml of diphenylamine solution R and,dropwise with shaking,5ml of nitrogen-free sulphuric acid R.Transfer the tubeto a water-bath at50°C.After15min,any blue colour in the solution is not more intense than that in a reference solution prepared at the same time in the same manner using a mixture of4.5ml of nitrate-free water R and0.5ml of nitrate standard solution(2ppm NO3)R.Aluminium(2.4.17):maximum10ppb,if intended for use in the manufacture of dialysis solutions.Prescribed solution.To400ml of the water to be examined add10ml of acetate buffer solution pH6.0R and100ml of distilled water R.Reference solution.Mix2ml of aluminium standard solution(2ppm Al)R,10ml of acetate buffer solutionpH6.0R and98ml of distilled water R.Blank solution.Mix10ml of acetate buffer solutionpH6.0R and100ml of distilled water R.Heavy metals(2.4.8):maximum0.1ppm.Heat200ml in a glass evaporating dish on a water-bath until the volume is reduced to20ml.12ml of the concentrated solution complies with limit test A.Prepare the standard using10ml of lead standard solution(1ppm Pb)R. Bacterial endotoxins(2.6.14):less than0.25IU/ml,if intended for use in the manufacture of dialysis solutions without a further appropriate procedure for removal of bacterial endotoxins.LABELLINGThe label states,where applicable,that the substance is suitable for use in the manufacture of dialysis solutions.Purified water in containers DEFINITIONPurified water in bulk that has been filled and stored in conditions designed to assure the required microbiological quality.It is free from any added substances. CHARACTERSAppearance:clear and colourless liquid.TESTSIt complies with the tests prescribed in the section on Purified water in bulk and with the following additional tests. Acidity or alkalinity.To10ml,freshly boiled and cooled in a borosilicate glass flask,add0.05ml of methyl red solution R. The solution is not coloured red.To10ml add0.1ml of bromothymol blue solution R1.The solution is not coloured blue.Oxidisable substances.To100ml add10ml of dilute sulphuric acid R and0.1ml of0.02M potassium permanganate and boil for5min.The solution remains faintly pink.Chlorides.To10ml add1ml of dilute nitric acid R and 0.2ml of silver nitrate solution R2.The solution shows no change in appearance for at least15min.Sulphates.To10ml add0.1ml of dilute hydrochloric acid R and0.1ml of barium chloride solution R1.The solution shows no change in appearance for at least1h. Ammonium:maximum0.2ppm.To20ml add1ml of alkaline potassium tetraiodomercurate solution R.After5min,examine the solution down the vertical axis of the tube.The solution is not more intensely coloured than a standard prepared at the same time by adding1ml of alkaline potassium tetraiodomercurate solution R to a mixture of4ml of ammonium standard solution(1ppm NH4)R and16ml of ammonium-free water R.3214See the information section on general monographs(cover pages)。

克林霉素磷酸酯C18H34ClN2O8PS[24729-96-2]来源于发酵产品的半合成产品。

含量：无水计有效物质百分之95.0%~102.0%。

特征外观：白色或几乎白色，易吸水粉末。

溶解性：易溶于水，微溶于乙醇（96%），几乎不溶于亚甲基氯化物。

这表明多态性（5.9）。

鉴定首先识别的是A，D。

其次是B、C、DA：红外吸收光谱仪（2.2.24）。

准备工作：溴酸钾R的分层。

在2个独立的管道中，检验50mg的样品和50mg的克林霉素磷酸酯的标准品。

加水加热溶解至0.2ml。

在100-105 ℃2小时减压干燥蒸发至干。

对照：克林霉素磷酸酯标准品。

B. 二薄层色谱法（2.2.27）。

样品组：将20mg的待检验样品溶于甲醇，用甲醇稀释至10ml。

对照品组：将克林霉素磷酸酯标准品20mg溶于甲醇中，用甲醇稀释至10ml。

空白组：将盐酸林可霉素标准品10mg用方案a的方法测试。

材料：薄层硅胶板R。

流动相：冰醋酸R，水俄R，丁醇R （20:20:60 V/V/V ）。

粒径：5ul。

展开：超过12厘米。

干燥：在100-105 ℃下30min。

检测：点0.1%溴酸钾R溶液系统适用性：参考方案b： - 色谱中主要显示两个明显的斑点。

结果：在色谱中取得的主要点和类似的点，在参考方案a中获得的色谱颜色和大小。

C.取大约10毫克，溶于2ml的稀盐酸酸R，水浴3分钟。

添加4ml的碳酸钠溶液R和1ml2%硝酸钠溶液R。

用同样的方法制备克林霉素磷酸酯标准品做为对照。

与标准品相对应的颜色就是所测试的结果。

D.0.1ml样品溶于5ml的浓氢氧化钠溶液R和5mlR水，用回流冷凝器煮沸蒸馏90min，冷却后添加5ml硝酸R溶液，分成3份，15ml每份，其中还有氯化物R和废弃物。

过滤，取滤液进行硝酸盐反应。

试验溶液S：1.00g在水中溶解，如有必要，可加热溶解。

冷却稀释到25ml。

澄清度：透明（参考2.2.1）和无色（参考2.2.2，方法二）。

Lactic acid EUROPEAN PHARMACOPOEIA7.0Heavy metals (2.4.8):maximum 20ppm.1.0g complies with test C.Prepare the reference solution using 2mL of lead standard solution (10ppm Pb)R .Loss on drying(2.2.32):maximum 1.0per cent,determined on 1.000g by drying in an oven at 105°C at a pressure not exceeding 0.7kPa.Sulfated ash (2.4.14):maximum 0.1per cent,determined on 1.0g.ASSAY In order to avoid overheating in the reaction medium,mix thoroughly throughout and stop the titration immediately after the end-point has been reached.Dissolve 0.200g in a mixture of 10mL of anhydrous formic acid R and 40mL of acetic anhydride R .Titrate with 0.1M perchloric acid ,determining the end-point potentiometrically(2.2.20).1mL of 0.1M perchloric acid is equivalent to 36.49mg of C 19H 25ClN 2O 3.IMPURITIES Specified impurities :A,B.A.R =H:2-hydroxy-5-[1-hydroxy-2-[(1-methyl-3-phenylpropyl)-amino]ethyl]benzoic acid,B.R =CH 3:methyl 2-hydroxy-5-[1-hydroxy-2-[(1-methyl-3-phenylpropyl)amino]ethyl]benzoate.01/2008:0458LACTIC ACID Acidumlacticum C 3H 6O 3M r 90.1DEFINITIONMixture of 2-hydroxypropanoic acid,its condensation products,such as lactoyl-lactic acid and polylactic acids,and water.Theequilibrium between lactic acid and polylactic acids depends on the concentration and temperature.It is usually the racemate ((RS )-lactic acid).Content :88.0per cent m/m to 92.0per cent m/m of C 3H 6O 3.CHARACTERSAppearance :colourless or slightly yellow,syrupy liquid.Solubility :miscible with water and with ethanol (96per cent).IDENTIFICATION A.Dissolve 1g in 10mL of water R .The solution is strongly acidic (2.2.4).B.Relative density (2.2.5):1.20to 1.21.C.It gives the reaction of lactates (2.3.1).TESTS Solution S .Dissolve 5.0g in 42mL of 1M sodium hydroxide and dilute to 50mL with distilled water R .Appearance .The substance to be examined is not more intensely coloured than reference solution Y 6(2.2.2,Method II ).Ether-insolublesubstances .Dissolve 1.0g in 25mL of ether R .The solution is not more opalescent than the solvent used for the test.Sugars and other reducing substances .To 1mLof solution Sadd 1mL of 1M hydrochloric acid ,heat to boiling,allow tocool and add 1.5mL of 1M sodium hydroxide and 2mL of cupri-tartaricsolution R .Heat to boiling.No red or greenish precipitate is formed.Methanol (2.4.24):maximum 50ppm,if intended for use in the manufacture of parenteral preparations.Citric,oxalic and phosphoric acids .To 5mL of solution S add dilute ammonia R1until slightly alkaline (2.2.4).Add 1mL ofcalcium chloride solution R .Heat on a water-bath for 5min.Both before and after heating,any opalescence in the solution is not more intense than that in a mixture of 1mL of water R and 5mL of solution S.Sulfates (2.4.13):maximum 200ppm.Dilute 7.5mL of solution S to 15mL with distilled water R .Calcium (2.4.3):maximum 200ppm.Dilute 5mL of solution S to 15mL with distilled water R .Heavy metals (2.4.8):maximum 10ppm.12mL of solution S complies with limit test A.Prepare the reference solution using lead standard solution (1ppm Pb)R .Sulfated ash (2.4.14):maximum 0.1per cent,determined on1.0g.Bacterial endotoxins (2.6.14):less than 5IU/g,if intended for use in the manufacture of parenteral preparations without a further appropriate procedure for the removalof bacterialendotoxins.Before use,neutralise the test solution to pH 7.0-7.5with strong sodium hydroxide solution R and shake vigorously.ASSAYPlace 1.000g in a ground-glass-stoppered flask and add 10mL ofwater R and 20.0mL of 1M sodium hydroxide .Close the flaskand allow to stand for ing 0.5mL of phenolphthalein solution R as indicator,titrate with 1M hydrochloric acid untilthe pink colour is discharged.1mL of 1M sodium hydroxide is equivalent to 90.1mgof C 3H 6O 3.LABELLINGThe label states,where applicable,that the substance is suitablefor use in the manufacture of parenteral preparations.01/2008:1771(S )-LACTIC ACIDAcidum (S)-lacticumC 3H 6O 3M r 90.1DEFINITIONMixture of (S )-2-hydroxypropanoic acid,its condensationproducts,such as lactoyl-lactic acid and polylactic acids,andwater.The equilibrium between lactic acid and polylactic acids depends on the concentration and temperature.Content:88.0per cent m/m to 92.0per cent m/m of C 3H 6O 3,not less than 95.0per cent of which is the (S )-enantiomer.CHARACTERSAppearance :colourless or slightly yellow,syrupyliquid.Solubility :miscible with water and with ethanol (96per cent).2328See the information section on general monographs (cover pages)EUROPEAN PHARMACOPOEIA 7.0Lactitolmonohydrate IDENTIFICATION A.Dissolve 1g in 10mL of water R .The solution is strongly acidic (2.2.4).B.Relative density (2.2.5):1.20to 1.21.C.It gives the reaction of lactates (2.3.1).D.It complies with the limits of the assay.TESTS Solution S .Dissolve 5.0g in 42mL of 1M sodium hydroxideand dilute to 50mL with distilled water R .Appearance .The substance to be examined is not more intensely coloured than reference solution Y 6(2.2.2,Method II ).Ether-insoluble substances .Dissolve 1.0g in 25mL of ether R .The solution is not more opalescent than the solvent used for the test.Sugars and other reducing substances .To 1mL of solution S add 1mL of 1M hydrochloric acid ,heat to boiling,allow to cool and add 1.5mL of 1M sodium hydroxide and 2mL of cupri-tartaric solution R .Heat to boiling.No red or greenishprecipitate is formed.Methanol (2.4.24):maximum 50ppm,if intended for use in themanufacture of parenteral preparations.Citric,oxalic and phosphoric acids .To 5mLof solution S add dilute ammoniaR1until slightly alkaline (2.2.4).Add 1mL ofcalcium chloride solution R .Heat on a water-bath for 5min.Both before and after heating,any opalescence in the solution is not more intense than that in a mixture of 1mL of water Rand 5mL of solution S.Sulfates (2.4.13):maximum 200ppm.Dilute 7.5mL of solution S to 15mL with distilled water R .Calcium (2.4.3):maximum 200ppm.Dilute 5mL of solution S to 15mL with distilled water R .Heavy metals (2.4.8):maximum 10ppm.12mL of solution S complies with limit test A.Prepare the reference solution using lead standard solution (1ppm Pb)R .Sulfated ash (2.4.14):maximum 0.1per cent,determined on 1.0g.Bacterial endotoxins (2.6.14):less than 5IU/g if intended foruse in the manufacture of parenteral preparations without a further appropriate procedure for the removal of bacterial endotoxins.Before use,neutralise the test solution to pH 7.0-7.5with strong sodium hydroxide solution R and shake vigorously.ASSAY Place 1.000g in a ground-glass-stoppered flask and add 10mL of water R and 20.0mL of 1M sodium hydroxide .Close the flask and allow to stand for ing 0.5mL of phenolphthalein solution R as indicator,titrate with 1M hydrochloric acid until the pink colour is discharged.1mL of 1M sodium hydroxide is equivalent to 90.1mg ofC 3H 6O 3.(S )-enantiomer Transfer an amount of the substance to be examined equivalent to 2.00g of lactic acid into a round-bottomed flask,add 25mL of 1M sodium hydroxide and boil gently for 15min.Cool down and adjust to pH 7.0using 1M hydrochloric acid .Add 5.0g of ammonium molybdate R ,dissolve and dilute to 50.0mL with water R .Filter and measure the angle of optical rotation (2.2.7).Calculate the percentage content of (S )-enantiomer using the expression:α=angle of optical rotation (absolute value),m=mass of the substance to be examined,in grams,c =percentage content of C 3H 6O 3in the substance tobe examined.Thecomplex of (S )-lactic acid formed under these test conditionsis BELLING The label states,where applicable,that the substance is suitablefor use in the manufacture of parenteral preparations.01/2009:1337corrected 6.5LACTITOL MONOHYDRATE LactitolummonohydricumC12H 24O 11,H 2O M r 362.3[81025-04-9]DEFINITION4-O -(β-D -Galactopyranosyl)-D -glucitol monohydrate.Content :96.5per cent to 102.0per cent (anhydrous substance).CHARACTERSAppearance :white or almost white,crystalline powder.Solubility:very soluble in water,slightly soluble in ethanol (96per cent),practically insoluble in methylene chloride IDENTIFICATIONFirst identification:B.Second identification:A,C.A.Specific optical rotation (see Tests).B.Infrared absorption spectrophotometry (2.2.24).Comparison :lactitol monohydrate CRS .C.Thin-layer chromatography (2.2.27).Testsolution.Dissolve 50mg of the substance to be examined in methanol R and dilute to 20mL with the samesolvent.Referencesolution (a).Dissolve 5mg of lactitol monohydrate CRS in methanol R and dilute to 2mL with the same solvent.Reference solution(b).Dissolve 2.5mg of sorbitol CRS(impurityE)in 1mL of reference solution (a)and dilute to10mL with methanol R .Plate :TLC silica gel G plate R .Mobile phase :water R ,acetonitrile R (25:75V/V ).Application :2μL.Development :over 2/3of the plate.Drying :in air.Detection:spray with 4-aminobenzoic acid solution R and dry in a current of cold air until the solvent is removed;heatat100°C for 15min and allow to cool;spray with a 2g/L solution of sodium periodate R and dry in a current of cold air;heat at 100°C for 15min.System suitabilit y:the chromatogram obtained withreference solution (b)shows 2clearly separated spots.General Notices (1)apply to all monographs and other texts 2329。

Pemetrexed Disodium HeptahydrateGeneral Notices(Ph. Eur. monograph 2637)C20H19N5Na2O6,7H2O 597.5 357166-29-1Action and useThymidylate synthetase inhibitor; cytostatic.Ph EurDEFINITIONDisodium(2S)-2-[[4-[2-(2-amino-4-oxo-4,7-dihydro-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]amino]pentanedio ate heptahydrate.Content97.5 per cent to 102.0 per cent (anhydrous substance).CHARACTERSAppearanceWhite or almost white powder.SolubilityFreely soluble in water, very slightly soluble in anhydrous ethanol, practically insoluble in methylene chloride.IDENTIFICATIONCarry out either tests A, C, D, E or tests B, C, D, E.Results The 1H NMR spectrum obtained is qualitatively similar to the 1H NMR spectrum obtainedwith pemetrexed disodium heptahydrate CRS; disregard the peak located at approximately 5.0 ppm for the comparison.C. It gives reaction (a) of sodium (2.3.1).D. Enantiomeric purity (see Tests).E. Water (see Tests).TESTSSolution SDissolve 0.56 g in carbon dioxide-free water R and dilute to 10.0 mL with the same solvent.Appearance of solutionSolution S is not more opalescent than reference suspension II (2.2.1) and not more intensely coloured than reference solution GY4 or Y4(2.2.2, Method II).pH (2.2.3)7.5 to 8.4 for solution S.Enantiomeric purityLiquid chromatography (2.2.29). Prepare the solutions immediately before use or store them at 2-8 °C for not more than 24 h.Solution A Dissolve 8 g of β-cyclodextrin R in 900 mL of water for chromatography R. Add 15 mLof triethylamine R then 6 mL of phosphoric acid R and adjust to pH 6.0 with phosphoric acid R. Dilute to 1000 mL with water for chromatography R.Test solution Dissolve 12 mg of the substance to be examined in water for chromatography R and dilute to 50.0 mL with the same solvent.Reference solution (a) Dissolve 6 mg of pemetrexed for system suitability CRS (containing impurity E) in water for chromatography R and dilute to 25.0 mL with the same solvent.Reference solution (b) Dilute 1.0 mL of the test solution to 100.0 mL with water for chromatography R.Dilute 3.0 mL of this solution to 10.0 mL with water for chromatography R.Column:∙—size: l = 0.25 m, Ø = 4.6 mm;∙—stationary phase: octadecylsilyl silica gel for chromatography R (5 µm) with a pore size of 12 nm; ∙—temperature: 40 °C.Mobile phase acetonitrile R, solution A (5:95 V/V).Flow rate 1.0 mL/min.Detection Spectrophotometer at 230 nm.Injection 50 µL.Run time 1.5 times the retention time of pemetrexed.Relative retention With reference to pemetrexed (retention time = about 30 min):impurity E = about 0.94.System suitability:∙—symmetry factor: maximum 2.0 for the principal peak in the chromatogram obtained with reference solution (b);∙—peak-to-valley ratio: minimum 5.0, where H p = height above the baseline of the peak due to impurity E and H v = height above the baseline of the lowest point of the curve separating this peak from the peak due to pemetrexed in the chromatogram obtained with reference solution (a).Calculation of percentage contents:∙— for impurity E, use the concentration of pemetrexed disodium heptahydrate in reference solution (b).Limit:∙—impurity E: maximum 0.3 per cent.Column rinse The following program is given for information only.Use a gradient column rinse before column storage or after 30 sample injections to avoid build-up on the column. If a drifting baseline is observed, allow additional time for equilibration with the mobile phase. If a blank chromatogram exhibits broad humps, perform a gradient column rinse.Rinsing solution A water for chromatography R.Rinsing solution B acetonitrile R1.Related substancesLiquid chromatography (2.2.29). Prepare the solutions immediately before use or store them at 2-8 °C for not more than 24 h.Solution A 1.45 g/L solution of ammonium formate R in water for chromatography R, adjusted to pH 3.5 with anhydrous formic acid R.Test solution Dissolve 20 mg of the substance to be examined in water for chromatography R and dilute to 100.0 mL with the same solvent.Reference solution (a) Dilute 1.0 mL of the test solution to 100.0 mL with water for chromatography R.Dilute 1.0 mL of this solution to 10.0 mL with water for chromatography R.Reference solution (b) In order to prepare impurities B and C in situ, dissolve 30 mg of the substance to be examined in 10.0 mL of a 4.0 g/L solution of sodium hydroxide R, heat at 70 °C for 40 minutes and allow to cool. Dilute 1.0 mL of the solution to 10.0 mL with water for chromatography R.Reference solution (c) Dissolve the contents of a vial of pemetrexed impurity mixture CRS (impurities A and D) in 1.0 mL of water for chromatography R.Column:∙—size: l = 0.15 m, Ø = 4.6 mm;∙—stationary phase: base-deactivated octylsilyl silica gel for chromatography R (3.5 µm).Mobile phase:∙—mobile phase A: acetonitrile R, solution A (5:95 V/V);∙—mobile phase B: acetonitrile R, solution A (30:70 V/V);Flow rate 1.0 mL/min.Detection Spectrophotometer at 250 nm.Injection 20 µL.Identification of impurities Use the chromatogram supplied with pemetrexed impurity mixture CRS and the chromatogram obtained with reference solution (c) to identify the peaks due to impurities A and D;use the chromatogram obtained with reference solution (b) to identify the peaks due to impurities B andC.Relative retention With reference to pemetrexed (retention time = about 26 min): impurity A = about 0.82;impurity B = about 0.87; impurity C = about 0.88; impurity D = about 0.90.System suitability Reference solution (b):∙—peak-to-valley ratio: minimum 1.5, where H p = height above the baseline of the peak due to impurity B and H v = height above the baseline of the lowest point of the curve separating this peak from the peak due to impurity C.Calculation of percentage contents:∙— for each impurity, use the concentration of pemetrexed disodium heptahydrate in reference solution(a).Limits:∙—impurities A, D: for each impurity, maximum 0.15 per cent;∙—unspecified impurities: for each impurity, maximum 0.10 per cent;∙—total: maximum 0.6 per cent;∙—reporting threshold: 0.05 per cent.Heavy metals (2.4.8)Maximum 20 ppm.Solvent mixture acetone R, water R (40:60 V/V).0.250 g complies with test H. Prepare the reference solution using 0.5 mL of lead standard solution (10ppm Pb) R.Water (2.5.12)19.5 per cent to 22.1 per cent, determined on 0.050 g.Bacterial endotoxins (2.6.14)Less than 0.17 IU/mg.ASSAYLiquid chromatography (2.2.29). Prepare the solutions immediately before use or store them at 2-8 °C for not more than 24 h.Acetate buffer Mix 1.7 mL of glacial acetic acid R and 900 mL of water for chromatography R, adjust to pH 5.3 with a 760 g/L solution of sodium hydroxide R in water for chromatography R and dilute to 1000 mL with water for chromatography R.Test solution Dissolve 30.0 mg of the substance to be examined in water for chromatography R and dilute to 200.0 mL with the same solvent.Reference solution Dissolve 30.0 mg of pemetrexed disodium heptahydrate CRS in water forchromatography R and dilute to 200.0 mL with the same solvent.Column:∙—size: l = 0.15 m, Ø = 4.6 mm;∙—stationary phase: base-deactivated octylsilyl silica gel for chromatography R (3.5 µm); ∙—temperature: 30 °C.Mobile phase acetonitrile R, acetate buffer (11:89 V/V).Flow rate 2.0 mL/min.Detection Spectrophotometer at 285 nm.Injection 20 µL.Run time Twice the retention time of pemetrexed (retention time = about 3 min).Calculate the percentage content of C20H19N5Na2O6 taking into account the assigned contentof pemetrexed disodium heptahydrate CRS.IMPURITIESSpecified impurities A, D, E.Other detectable impurities (the following substances would, if present at a sufficient level, be detected by one or other of the tests in the monograph. They are limited by the general acceptance criterion for other/unspecified impurities and/or by the general monograph Substances for pharmaceutical use (2034).It is therefore not necessary to identify these impurities for demonstration of compliance. See also 5.10.Control of impurities in substances for pharmaceutical use): B, C.A.(2S)-2-[[4-[2-(2-amino-1-methyl-4-oxo-4,7-dihydro-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]amino ]-pentanedioic acid,B.(2S,2′S)-2,2′-[[(5R)-2,2′-diamino-4,4′,6-trioxo-1,4,4′,6,7,7′-hexahydro-1′H,5H-5,6′-bipyrrolo[2,3-d]pyrimi dine-5,5′-diyl]bis(ethylenebenzene-4,1-diylcarbonylimino)]dipentanedioic acid,C.(2S,2′S)-2,2′-[[(5S)-2,2′-diamino-4,4′,6-trioxo-1,4,4′,6,7,7′-hexahydro-1′H,5H-5,6′-bipyrrolo[2,3-d]pyrimi dine-5,5′-diyl]bis(ethylenebenzene-4,1-diylcarbonylimino)]dipentanedioic acid,D.(2S)-2-[[(4S)-4-[[4-[2-(2-amino-4-oxo-4,7-dihydro-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]amino]-4-carboxybutanoyl]amino]pentanedioic acid,E.(2R)-2-[[4-[2-(2-amino-4-oxo-4,7-dihydro-1H-pyrrolo[2,3-d]pyrimidin-5-yl)ethyl]benzoyl]amino]pentane dioic acid.Ph Eur。

EUROPEAN PHARMACOPOEIA 6.0ParenteralpreparationsMucoadhesive preparations DEFINITION Mucoadhesive preparations contain one or more active substances intended for systemic absorption through the buccal mucosa over a prolongedperiodof time.They may be supplied as mucoadhesive buccal tablets or as other mucoadhesive solid or semi-solid preparations.Mucoadhesive buccal tablets are prepared by compressionof mono-or multi-layeredtablets.They usually containhydrophilic polymers,which on wetting with the salivaproduce a flexible hydrogel that adheres to the buccal mucosa.PRODUCTION In the manufacture of mucoadhesive buccal tablets,measures are taken to ensure that they possess suitablemechanical strength to resist handling without crumbling or breaking.This may bedemonstratedby examining the Friability of uncoated tablets (2.9.7)and the Resistance to crushing of tablets (2.9.8).TESTS Dissolution .Unlessotherwisejustified and authorised,a suitable test is carried out to demonstrate the appropriate release of the active substance(s).01/2008:0520PARENTERAL PREPARATIONS Parenteralia The requirementsof this monograph do not necessarily apply to products derived fromhuman blood,toimmunological preparations,or radiopharmaceutical preparations.Special requirements may apply to preparations for veterinary use depending on the species of animal for which the preparation is intended.DEFINITION Parenteral preparations are sterilepreparations intended for administration by injection,infusion or implantation into the human or animal body.Parenteral preparations may require the use of excipients,for example to make the preparation isotonic with respect to blood,to adjustthe pH,to increase solubility,to prevent deterioration of the activesubstancesor to provideadequateantimicrobial properties,but not to adversely affect the intended medicinal action of the preparation or,at the concentrations used,to cause toxicity or undue local irritation.Containers for parenteral preparations are made as far as possible frommaterialsthat are sufficiently transparentto permit the visual inspection of the contents,except for implants and in other justified and authorised cases.Where applicable,the containers for parenteral preparations comply with the requirements for Materials used for the manufacture of containers (3.1and subsections)and Containers (3.2and subsections).Parenteral preparations are supplied in glass containers (3.2.1)or in other containers such as plastic containers (3.2.2,3.2.2.1and 3.2.9)and prefilled syringes.The tightness of the containeris ensured bysuitablemeans.Closuresensure a good seal,prevent the access of micro-organisms and other contaminants and usually permit the withdrawal ofa part or the whole of the contents without removal of the closure.The plasticmaterials or elastomers (3.2.9)used to manufacture the closures are sufficiently firm and elastic to allow the passage of a needle with the least possible shedding ofparticles.Closures for multidose containers aresufficiently elastic to ensure that the puncture is resealedwhen the needle is withdrawn.Severalcategories of parenteral preparations may be distinguished:—injections,—infusions,—concentrates for injections or infusions,—powders for injections or infusions,—gels for injections,—implants.PRODUCTIONDuringthe developmentof a parenteral preparation,theformulation for which contains an antimicrobial preservative,the effectiveness of the chosen preservative shall be demonstratedto the satisfaction of the competent authority.A suitable test method togetherwithcriteria forjudgingthe preservative properties of the formulation are provided under Efficacy of antimicrobial preservation (5.1.3).Parenteral preparations are prepared using materials and methods designed to ensure sterility and to avoidthe introduction of contaminants and thegrowth ofmicro-organisms.Recommendations on this aspect areprovided in the text on Methods of preparation of sterile products (5.1.1).Water used in the manufacture of parenteral preparationscomplies with the requirements of water for injections in bulk stated in the monograph on Water for injections (0169).TESTSParticulate contamination:sub-visible particles (2.9.19).For preparations for human use,solutions for infusion orsolutions for injection comply with the test.Inthe case of preparations for subcutaneous or intramuscular injection,higher limits may be appropriate.Radiopharmaceutical preparations are exempt from these requirements.Preparations for which the label states thatthe product is to be used with a final filter are exempt from these requirements,providing it has been demonstrated that the filter delivers a solution that complies with the test.For preparations for veterinary use,whensupplied in containers with a nominal content of more than 100ml and when the content is equivalent to a dose of more than1.4ml per kilogram of body mass,solutions for infusion orsolutions for injection comply with the test for particulate contamination:sub-visible particles.Sterility (2.6.1).Parenteral preparations comply with the test for sterility.STORAGEIn a sterile,airtight,tamper-proof container.LABELLINGThe label states:—the name andconcentration of any added antimicrobial preservative,—where applicable,that the solution is to be used in conjunction with a final filter,—where applicable,that the preparation is free frombacterial endotoxins or that it is apyrogenic.General Notices (1)apply to all monographs and other texts 735Parenteral preparations EUROPEAN PHARMACOPOEIA6.0InjectionsDEFINITIONInjections are sterile solutions,emulsions or suspensions. They are prepared by dissolving,emulsifying or suspending the active substance(s)and any added excipients in water,in a suitable non-aqueous liquid,that may be non-sterile where justified,or in a mixture of these vehicles.Solutions for injection,examined under suitable conditions of visibility,are clear and practically free from particles. Emulsions for injection do not show any evidence of phase separation.Suspensions for injection may show a sediment which is readily dispersed on shaking to give a suspension which remains sufficiently stable to enable the correct dose to be withdrawn.Multidose preparations.Multidose aqueous injections contain a suitable antimicrobial preservative at an appropriate concentration except when the preparation itself has adequate antimicrobial properties.When a preparation for parenteral use is presented in a multidose container, the precautions to be taken for its administration and more particularly for its storage between successive withdrawals are given.Antimicrobial preservatives.Aqueous preparations which are prepared using aseptic precautions and which cannot be terminally sterilised may contain a suitable antimicrobial preservative in an appropriate concentration.No antimicrobial preservative is added when:—the volume to be injected in a single dose exceeds15ml, unless otherwise justified,—the preparation is intended for administration by routes where,for medical reasons,an antimicrobial preservative is not acceptable,such as intracisternally,epidurally,intrathecally or by any route giving access to thecerebrospinal fluid,or intra-or retro-ocularly.Such preparations are presented in single-dose containers.PRODUCTIONIn the manufacture of injections containing dispersed particles,measures are taken to ensure a suitable and controlled particle size with regard to the intended use. Single-dose preparations.The volume of the injection in a single-dose container is sufficient to permit the withdrawal and administration of the nominal dose using a normal technique(2.9.17).TESTSUniformity of dosage units.Single-dose suspensions for injection comply with the test for uniformity of dosage units (2.9.40)or,where justified and authorised,with the test for uniformity of content shown below.Herbal drugs and herbal drug preparations present in the dosage form are not subject to the provisions of this paragraph.Uniformity of content(2.9.6).Unless otherwise prescribed or justified and authorised,single-dose suspensions for injection with a content of active substance less than2mg or less than2per cent of the total mass comply with test A for uniformity of content of single-dose preparations.Ifthe preparation contains more than one active substance, the requirement applies only to those substances that correspond to the above conditions.Bacterial endotoxins-pyrogens.A test for bacterial endotoxins(2.6.14)is carried out or,where justified and authorised,the test for pyrogens(2.6.8).Recommendations on the limits for bacterial endotoxins are given in chapter2.6.14.Preparations for human use.The preparation complies with a test for bacterial endotoxins(2.6.14)or with the test for pyrogens(2.6.8).Preparations for veterinary use.When the volume to be injected in a single dose is15ml or more and is equivalent to a dose of0.2ml or more per kilogram of body mass,the preparation complies with a test for bacterial endotoxins (2.6.14)or with the test for pyrogens(2.6.8).Any preparation.Where the label states that the preparation is free from bacterial endotoxins or apyrogenic,respectively, the preparation complies with a test for bacterial endotoxins (2.6.14)or with the test for pyrogens(2.6.8),respectively.InfusionsDEFINITIONInfusions are sterile,aqueous solutions or emulsions with water as the continuous phase.They are usually made isotonic with respect to blood.They are principally intended for administration in large volume.Infusions do not contain any added antimicrobial preservative.Solutions for infusion,examined under suitable conditions of visibility are clear and practically free from particles. Emulsions for infusion do not show any evidence of phase separation.PRODUCTIONIn the manufacture of infusions containing dispersed particles,measures are taken to ensure a suitable and controlled particle size with regard to the intended use. The volume of the infusion in the container is sufficientto permit the withdrawal and administration of the nominal dose using a normal technique(2.9.17).TESTSBacterial endotoxins-pyrogens.They comply with a test for bacterial endotoxins(2.6.14)or,where justified and authorised,with the test for pyrogens(2.6.8).For the latter test inject10ml per kilogram of body mass into each rabbit, unless otherwise justified and authorised. Concentrates for injections or infusions DEFINITIONConcentrates for injections or infusions are sterile solutions intended for injection or infusion after dilution.They are diluted to a prescribed volume with a prescribed liquid before administration.After dilution,they comply with the requirements for injections or for infusions.TESTSBacterial endotoxins-pyrogens.They comply with the requirements prescribed for injections or for infusions,after dilution to a suitable volume.Powders for injections or infusions DEFINITIONPowders for injections or infusions are solid,sterile substances distributed in their final containers and which,when shaken with the prescribed volume of a prescribed sterile liquid rapidly form either clear and736See the information section on general monographs(cover pages)EUROPEAN PHARMACOPOEIA 6.0Patches,transdermal practically particle-free solutions or uniform suspensions.After dissolution or suspension,they comply with therequirements for injections or for infusions.Freeze-dried products for parenteral use are considered aspowders for injections or infusions.PRODUCTION The uniformity of contentand uniformityof mass offreeze-dried products for parenteral use are ensured by the in-process control of the amount of the solution prior to freeze-drying.TESTS Uniformity of dosage units .Powders for injections or infusions comply with the test for uniformity of dosage units (2.9.40)or,where justified and authorised,with the tests for uniformity of content and/or uniformity of mass shown below.Herbal drugs and herbal drug preparations present in the dosage form are not subject to the provisions of this paragraph.Uniformity of content (2.9.6).Unless otherwise prescribed or justifiedand authorised,powders for injections or infusions with a content of active substance less than 2mgor less than 2per cent of the total mass,or with a unit mass equal to or less than 40mg comply with test A for uniformity of content of single-dose preparations.If the preparation contains more than one active substance,the requirement applies only to those substances that correspond to the above conditions.Uniformity of mass (2.9.5).Powders for injections or infusions complywith the test for uniformity of mass ofsingle-dose preparations.If the test for uniformity of content is prescribed for all the active substances,the test for uniformity of mass is not required.Bacterial endotoxins-pyrogens.Theycomplywith the requirements prescribed for injections or for infusions,after dissolution or suspension in a suitable volume of BELLING The label states the instructions for the preparation of injections and infusions.Gels for injections DEFINITION Gels for injections are sterile gels with a viscosity suitable to guarantee a modified release of the active substance(s)at the site of injection.Implants DEFINITION Implants are sterile,solid preparations of a size and shape suitable for parenteral implantation and release of the active substance(s)over an extended period of time.Each dose is provided in a sterile container.01/2008:1011PATCHES,TRANSDERMALEmplastra transcutaneaDEFINITIONTransdermal patches are flexible pharmaceutical preparations of varying sizes,containing one or more activesubstances.Theyare intendedto be applied to the unbrokenskin in order to deliver the active substance(s)to the systemiccirculation after passing through the skin barrier.Transdermalpatches normally consist of an outer covering which supports a preparation which contains the activesubstance(s).The transdermal patches are covered on thesite of the release surface of the preparation by a protective liner,which is removed before applying the patch to the skin.The outer covering is a backing sheet impermeable to the active substance(s)and normally impermeable to water,designed to support and protect the preparation.The outer covering may have the same dimensions as the preparation or it may be larger.In the latter case the overlapping border of the outer covering is covered by pressure-sensitive adhesive substances which assure the adhesion of the patchto the skin.The preparation contains the active substance(s)togetherwith excipients such as stabilisers,solubilisers or substances intended to modify the release rate or to enhance transdermal absorption.It may be a single layer or multi-layer solid orsemi-solid matrix,and in this case it is the compositionand structure of the matrix which determines the diffusion pattern of the active substance(s)to the skin.The matrix may contain pressure-sensitive adhesives which assure theadhesion of the preparation to the skin.The preparation may exist as a semi-solid reservoir one side of which is amembranewhich may control the release and the diffusion of the active substance(s)from the preparation.The pressure-sensitive adhesive substances may,in this case,beapplied to some or all parts of the membrane,or only aroundthe border of the membrane of the outer covering.When applied to the dried,clean and unbrokenskin,thetransdermalpatch adheres firmly to the skin by gentle pressure of the hand or the fingers and can be peeled offwithout causing appreciable injury to the skin or detachment of thepreparation from the outer covering.The patch mustnot be irritant or sensitising to the skin,even after repeated applications.The protective liner generally consists of a sheet of plastic ormetal material.When removed,the protective liner does notdetach the preparation (matrix or reservoir)or the adhesive from the patch.Transdermal patches are normally individually enclosed in sealed sachets.PRODUCTIONInthe manufacture,packaging,storage and distributionof transdermalpatches suitable means are taken to ensuretheir microbial quality;recommendations onthis aspect are provided in the text on Microbiological quality ofpharmaceutical preparations (5.1.4).TESTSUniformity ofdosageunits .Transdermal patchescomplywith the test for uniformity of dosage units (2.9.40)or,where justified and authorised,with the test for uniformity General Notices (1)apply to all monographs and other texts 737。

EP5.1.10 细菌内毒素测试使用指南（中英文1/2）2015-12-28 22:58:13| 分类：EP07/2016: 501105.1.10. GUIDELINES FOR USING THE TEST FOR BACTERIAL ENDOTOXINS细菌内毒素测试使用指南1. INTRODUCTION 概述Endotoxins from gram-negative bacteria are the most common cause of toxic reactions resulting from contamination of pharmaceutical products with pyrogens; their common pyrogenic activity is much higher tha n that of other known pyrogenic substances. These endotoxins are lipopolysaccharides. Although there are a small number of pyrogens that possess a different structure, the conclusion is generally justified that the absence of bacterial endotoxins in a substance or product implies the absence of pyrogenic components, provided the presence of non-endotoxin pyrogenic substances can be ruled out. The monocyte-activation test (2.6.30) is a suitable method to use to rule out the presence of non-endotoxin pyrogens in substances or products.革兰氏阳性菌产生的内毒素是最常见的有毒性反应原因，其原因是药品受到热源污染。