PARENTERALPARENTERAL

PARENTERALPARENTERAL

PARA→OUTSIDEPARA→OUTSIDE ENTERON→INTESTINEENTERON→INTESTINE Denotes Route of administration other than Denotes Route of administration other than

oral routeoral route Refers to injectable routes administrationRefers to injectable routes administration Necessity ? Necessity ?

Poor Bioavailability & Narrow Poor Bioavailability & Narrow therapeutic index , Immediate physiologic therapeutic index , Immediate physiologic response ex: cardiac response ex: cardiac arrest, asthma & shockarrest, asthma & shock

oral VS parenteraloral VS parenteral

Oral Injectable

Rapid onset of Action √

Less expensive √

Administrable to non responsive patients

√

Patient convenience & comfort √

Administrable directly to site of action

√

Retrievable , if Necessary √

Better Absorption √

PARENTERAL ROUTESPARENTERAL ROUTES

Intra- articular → JointsIntra- articular → Joints Intra spinal → spinal columnIntra spinal → spinal column Intra arterial → arteriesIntra arterial → arteries Intravenous → veinsIntravenous → veins Intra dermal → skinIntra dermal → skin Intra synovial → joint fluidIntra synovial → joint fluid Intra thecal → spinal fluidIntra thecal → spinal fluid

PROS N CONSPROS N CONS

– Quick onset of actionQuick onset of action– Suitable for the drugs which are not Suitable for the drugs which are not

administered by oral routeadministered by oral route– Useful for unconscious or vomiting patients.Useful for unconscious or vomiting patients.– Duration of action can be prolonged by Duration of action can be prolonged by

modifying formulation.modifying formulation.– Suitable for nutritive like glucose & Suitable for nutritive like glucose &

electrolyte.electrolyte.– Suitable for the drugs which are inactivated Suitable for the drugs which are inactivated

in GIT or HCl (GI fluid)in GIT or HCl (GI fluid)

PROS N CONSPROS N CONS

– Once injected cannot be controlled Once injected cannot be controlled (retreat)(retreat)

– Injections may cause pain at the site of Injections may cause pain at the site of injectioninjection

– Only trained person is requiredOnly trained person is required– If given by wrong route, difficult to control If given by wrong route, difficult to control

adverse effectadverse effect– Difficult to save patient if overdoseDifficult to save patient if overdose– Sensitivity or allergic reaction at the site Sensitivity or allergic reaction at the site

of injectionof injection– Requires strict control of sterility & non Requires strict control of sterility & non

pyrogenicity than other formulation.pyrogenicity than other formulation.

Necessities of parenteral Necessities of parenteral preparationspreparations

– SterilitySterility (must)(must)

– PyrogenPyrogen (must)(must)

– Free from particulate matter Free from particulate matter

(must)(must)

– ClarityClarity (must)(must)

– StabilityStability (must)(must)

– IsotonicityIsotonicity (should)(should)

Necessities of parenteral Necessities of parenteral preparationspreparations

– Solvents or vehicles used must meet Solvents or vehicles used must meet

special purity and other standards.special purity and other standards.

– Restrictions on buffers, stabilizers, Restrictions on buffers, stabilizers,

antimicrobial preservative. Do not use antimicrobial preservative. Do not use

coloring agents.coloring agents.

– Must be prepared under aseptic conditions.Must be prepared under aseptic conditions.

– Specific and high quality packaging.Specific and high quality packaging.

PARENTERAL ROUTESPARENTERAL ROUTES

Intra cardiac → heartIntra cardiac → heart Intra muscular →musclesIntra muscular →muscles Subcutaneous → under the skinSubcutaneous → under the skin Intra cisternal → Brain Intra cisternal → Brain

ventriclesventricles Intra peritonial → StomachIntra peritonial → Stomach Intra plueral → LungsIntra plueral → Lungs

PARENTERAL TYPESPARENTERAL TYPES

Powder for Injection Powder for Injection Collidal solutionCollidal solution Injectable emulsionInjectable emulsion Injectable suspensionInjectable suspension Oily injection (solution)Oily injection (solution) Infusion fluidInfusion fluid

Formulation of parenteralsFormulation of parenterals

1.1. Therapeutic agentsTherapeutic agents2.2. VehiclesVehicles

i.i. WaterWaterii.ii. Water miscible vehiclesWater miscible vehiclesiii.iii.Non- aqueous vehiclesNon- aqueous vehicles

3.3. Added substances (Additives)Added substances (Additives)i.i. AntimicrobialsAntimicrobialsii.ii. AntioxidantsAntioxidantsiii.iii.BuffersBuffersiv.iv.Bulking agentsBulking agents

Formulation of parenteralsFormulation of parenterals

i.i. Chelating agentsChelating agentsii.ii. ProtectantsProtectantsiii.iii.Solubilizing agentsSolubilizing agentsiv.iv.SurfactantsSurfactantsv.v. Tonicity- adjusting agentsTonicity- adjusting agents

General steps involved General steps involved 1. Cleaning

2. Preparation of bulk products

3. Filtration

4. Filling of solution in or product in ampoule or vial

7. Tests for Quality control

5.Sealing

6. Sterilization

Formulation of ParenteralFormulation of Parenteral

1.Therapeutic ingredients:1.Therapeutic ingredients: InsulinInsulin AntibioticsAntibiotics AnticancerAnticancer SteroidsSteroids Vaccines Vaccines AntipyreticAntipyretic AnalgesicsAnalgesics Anti- inflammatory Anti- inflammatory LVP’s like Dextrose, NaCl or combination etc….LVP’s like Dextrose, NaCl or combination etc….

Formulation of ParenteralFormulation of Parenteral

2.Solvents: 2.Solvents:

o Water Water

o Should meet compendial requirementsShould meet compendial requirements

o Water miscible vehiclesWater miscible vehicles

o Ethyl alcoholEthyl alcohol

o PEGPEG

o PGPG

o Non aqueous vehicles Non aqueous vehicles

o Fixed oilsFixed oils

Formulation of ParenteralFormulation of Parenteral

SolventsSolventsSolvents used must be:Solvents used must be: Non-irritatingNon-irritating Non-toxicNon-toxic Non-sensitizingNon-sensitizing No pharmacological activity of its No pharmacological activity of its

ownown Not affect activity of medicinalNot affect activity of medicinal

Formulation of ParenteralFormulation of Parenteral

3. 3. Added substances (Additives)Added substances (Additives)– Antimicrobials: Antimicrobials:

Added for fungi static or bacteriostatic action or Added for fungi static or bacteriostatic action or concentrationconcentration

Used to prevent the multiplication of micro-organismsUsed to prevent the multiplication of micro-organisms Ex..Ex..

– Benzyl alcohol ------Benzyl alcohol ------ 0.5 – 10 %0.5 – 10 %– Benzethonium chloride --Benzethonium chloride -- 0.01 % 0.01 % – Methyl paraben ----Methyl paraben ---- 0.01 – 0.18 % 0.01 – 0.18 % – Propyl paraben ---Propyl paraben --- 0.005 – 0.035 % 0.005 – 0.035 % – Phenol ---Phenol --- 0.065 – 0.5 %0.065 – 0.5 %

Formulation of ParenteralFormulation of Parenteral

PreservativesPreservatives:: Multidose Multidose containers must have containers must have preservatives unless prohibited preservatives unless prohibited by monograph.by monograph.

Large volume parenteral must Large volume parenteral must not contain preservative becoz not contain preservative becoz it may be dangerous to human it may be dangerous to human body if it contain in high doses.body if it contain in high doses.

Formulation of ParenteralFormulation of Parenteral– Antioxidants:Antioxidants:

Used to protect product from oxidation Used to protect product from oxidation Acts as reducing agent or prevents oxidationActs as reducing agent or prevents oxidation Ex: Ex:

– A) Reducing agent:A) Reducing agent: Ascorbic acid --Ascorbic acid -- 0.02 – 0.1 %0.02 – 0.1 % Sodium bisulphite--Sodium bisulphite-- 0.1 – 0.15 %0.1 – 0.15 % Sodium metabisulphite--Sodium metabisulphite-- 0.1 – 0.15 % 0.1 – 0.15 % Thiourea -Thiourea - 0.005 %0.005 %

– B) B) Blocking agentsBlocking agents:: Ascorbic acid esters-Ascorbic acid esters- 0.01 – 0.015% 0.01 – 0.015% BHT-BHT- 0.005 – 0.02 %0.005 – 0.02 %

– C) C) Synergistic:Synergistic: Ascorbic acid , Citric acid , Tartaric acid Ascorbic acid , Citric acid , Tartaric acid

– D) D) Chelating agent:Chelating agent: EDTA-EDTA- 0.01- 0.075 %0.01- 0.075 %

Formulation of ParenteralFormulation of Parenteral

Buffers:Buffers:

– Added to maintain pH,Added to maintain pH,

– Change in pH may causes degradation of the products Change in pH may causes degradation of the products

– Acetates, citrates, phosphates are generally used.Acetates, citrates, phosphates are generally used. Factors affecting selection of buffers:Factors affecting selection of buffers:

Effective range,Effective range, Concentration Concentration Chemical effect on the total productChemical effect on the total product

EXAMPLESEXAMPLES::

– Acetic acid ,adipic acid, benzoic acid, citric acid, lactic acid Acetic acid ,adipic acid, benzoic acid, citric acid, lactic acid

– Used in the conc. of 0.1 to 5.0 % Used in the conc. of 0.1 to 5.0 %

Formulation of ParenteralFormulation of Parenteral

Chelating agents:Chelating agents:

– Used to form the complex with the metallic ions present in the Used to form the complex with the metallic ions present in the formulation so that the ions will not interfere during mfg. of formulation so that the ions will not interfere during mfg. of formulation.formulation.

– They form a complex which gets dissolved in the solvents.They form a complex which gets dissolved in the solvents. Examples:Examples:

– Disodium edetate – 0.00368 - .05 % Disodium edetate – 0.00368 - .05 %

– Disodium calcium edetate - 0.04 %Disodium calcium edetate - 0.04 %

– Tetrasodium edetate – 0.01 % Tetrasodium edetate – 0.01 %

Formulation of ParenteralFormulation of Parenteral

Stabilizers:Stabilizers: – As parenterals are available in solution form they are most As parenterals are available in solution form they are most

prone to unstabilizeprone to unstabilize – Used to stabilize the formulation Used to stabilize the formulation – Maintain stable Maintain stable

Examples:Examples:– Creatinine – 0.5- 0.8 % Creatinine – 0.5- 0.8 % – Glycerin – 1.5 – 2.25 % Glycerin – 1.5 – 2.25 % – Niacinamide – 1.25 -2.5 % Niacinamide – 1.25 -2.5 % – Sodium saccharin – 0.03 %Sodium saccharin – 0.03 %

Formulation of ParenteralFormulation of Parenteral Solubilizing agents:Solubilizing agents:

Used to increase solubility of slightly soluble drugsUsed to increase solubility of slightly soluble drugs they acts by any one of the following: they acts by any one of the following: solubilizers, solubilizers, emulsifiers or emulsifiers or wetting agents.wetting agents.

Examples:Examples: Dimethylacetamide,Dimethylacetamide, Ethyl alcoholEthyl alcohol GlycerinGlycerin LecithinLecithin PEG – 40 castor oil PEG – 40 castor oil PEG – 300 PEG – 300 Polysorbate 20, 40, 80 Polysorbate 20, 40, 80

Formulation of ParenteralFormulation of Parenteral Tonicity- adjusting agents:Tonicity- adjusting agents:

– Used to reduce the pain of injection.Used to reduce the pain of injection.– Buffers may acts as tonicity contributor as well as Buffers may acts as tonicity contributor as well as

stabilizers for the pH.stabilizers for the pH.– Isotonicity depends on permeability of a living Isotonicity depends on permeability of a living

semipermaeable membranesemipermaeable membrane Hypotonic : swelling of cells (enlargement)Hypotonic : swelling of cells (enlargement) Hypertonic: shrinking of cells (reduction) Hypertonic: shrinking of cells (reduction)

ExampleExample::– Glycerin Glycerin – Lactose Lactose – Mannitol Mannitol – Dextrose Dextrose – Sodium chloride Sodium chloride – SorbitolSorbitol

LABELLINGLABELLING– Name of productName of product– Quantity of the productQuantity of the product– % of drug or amount of drug in specified volume % of drug or amount of drug in specified volume

of amount of drug and volume of liquid to be of amount of drug and volume of liquid to be added added

– Name and quantity of all added substancesName and quantity of all added substances– Mfg. license no.Mfg. license no.– Batch no.Batch no.– Manufacturer/DistributorManufacturer/Distributor– Mfg. & Expiration dateMfg. & Expiration date– Retail price (incl. of all taxes)Retail price (incl. of all taxes)– Mfger. addressMfger. address– Veterinary product should be so labeledVeterinary product should be so labeled

LABELLINGLABELLING

Must check each individual monogram for:Must check each individual monogram for:– Type of container:Type of container:

Glass Glass Plastic Plastic Rubber closure Rubber closure

– Type of glassType of glass– Type I Type I – Type IIType II– Type III Type III – NP NP

– Tests for glass containers Tests for glass containers Powdered Glass test Powdered Glass test Water Attack test Water Attack test

– Package sizePackage size– Special storage instructionsSpecial storage instructions

Mixer/HomogenizerMixer/Homogenizer

Filtration assemblyFiltration assembly

Bottling/Filling Bottling/Filling machinerymachinery

PERSONNELPERSONNEL

Clothing of appropriate quality:Clothing of appropriate quality:– Grade DGrade D

hair, beard, moustache coveredhair, beard, moustache covered Protective clothing and shoesProtective clothing and shoes

– Grade CGrade C Hair, beard, moustache coveredHair, beard, moustache covered Single or 2-piece suit (covering wrists, high neck), shoesSingle or 2-piece suit (covering wrists, high neck), shoes no fibres to be shedno fibres to be shed

– Grade A and BGrade A and B Headgear, beard and moustache covered, masks, glovesHeadgear, beard and moustache covered, masks, gloves No shedding of fibres, and retain particles shed by operatorsNo shedding of fibres, and retain particles shed by operators

Production facilities: Production facilities:

– Clean- up areaClean- up area

– Preparation area Preparation area

– Aseptic area Aseptic area

– Quarantine area Quarantine area

– Finishing and packaging areaFinishing and packaging area

Sterile area Sterile area

LAY OUTLAY OUT

STOCK ROOM

COMPOUNDING AREA

CLEAN UP AREA

ASEPTIC AREA

QUARANTINE AREA

STERILIZATION

STORAGE

AND

TRANSPORT

PACKING

AND

LABELLING

Clean- up area:Clean- up area:– Non aseptic area Non aseptic area

– Free from dust ,fibres & micro-organismsFree from dust ,fibres & micro-organisms

– Constructed in such a way that should withstand Constructed in such a way that should withstand

moisture, steam & detergentmoisture, steam & detergent

– Ceiling & walls are coated with material to prevent Ceiling & walls are coated with material to prevent

accumulation of dust & micro-organismsaccumulation of dust & micro-organisms

– Exhaust fans are fitted to remove heat & humidityExhaust fans are fitted to remove heat & humidity

– The area should be kept clean so that to avoid The area should be kept clean so that to avoid

contamination to aseptic areacontamination to aseptic area

– The containers & closures are washed & dried in The containers & closures are washed & dried in

this area.this area.

Preparation area: Preparation area: – The ingredients are mixed & preparation is The ingredients are mixed & preparation is

prepared for filling prepared for filling

– Not essential that the area is asepticNot essential that the area is aseptic

– Strict precaution is taken to prevent Strict precaution is taken to prevent

contamination from outsidecontamination from outside

– Cabinets & counters: SS Cabinets & counters: SS

– Ceiling & walls : sealed & paintedCeiling & walls : sealed & painted

Aseptic area: Aseptic area:

– Filtration & filling into final containers & sealing is Filtration & filling into final containers & sealing is done done

– The entry of outside person is strictly prohibited The entry of outside person is strictly prohibited – To maintain sterility, special trained persons are To maintain sterility, special trained persons are

only allowed to enter & work only allowed to enter & work – Person who worked should wear sterile cloths Person who worked should wear sterile cloths – Should be subjected for physical examination to Should be subjected for physical examination to

ensure the fitness ensure the fitness – Minimum movement should be there in this area Minimum movement should be there in this area – Ceiling & walls & floors : sealed & painted or Ceiling & walls & floors : sealed & painted or

treated with aseptic solution and there should not treated with aseptic solution and there should not be any toxic effect of this treatment be any toxic effect of this treatment

Aseptic area: Aseptic area:

– Cabinets & counters: SSCabinets & counters: SS– Mechanical equipments : SS Mechanical equipments : SS – AIR: AIR:

Free from fibres, dust & micro organismsFree from fibres, dust & micro organisms HEPA filters are used which removes particles HEPA filters are used which removes particles

upto 0.3 micron upto 0.3 micron Fitted in laminar air flow system, in which air is Fitted in laminar air flow system, in which air is

free from dust & micro organisms flows with free from dust & micro organisms flows with uniform velocityuniform velocity

Air supplied is under positive pressure which Air supplied is under positive pressure which prevents particulate contamination from sweeping prevents particulate contamination from sweeping

UV lamps are fitted to maintain sterilityUV lamps are fitted to maintain sterility

Quarantine area: Quarantine area:

– After filling, sealing & sterilization the After filling, sealing & sterilization the products or batch is kept in this areaproducts or batch is kept in this area

– The random samples are chosen and given The random samples are chosen and given for analysis to QC dept. for analysis to QC dept.

– The batch is send to packing after issuing The batch is send to packing after issuing satisfactory reports of analysis from QCsatisfactory reports of analysis from QC

– If any problem is observed in above analysis If any problem is observed in above analysis the decision is to be taken for reprocessing or the decision is to be taken for reprocessing or others..others..

Finishing and packaging Finishing and packaging area: area:

– After proper label, the product is given for After proper label, the product is given for packingpacking

– Packing is done to protect the product Packing is done to protect the product from external environment from external environment

– The ideal Packing is that which protects The ideal Packing is that which protects the product during transportation, storage, the product during transportation, storage, shipping & handling.shipping & handling.

– The labeled container should be packed in The labeled container should be packed in cardboard or plastic containerscardboard or plastic containers

– Ampoules should be packed in partitioned Ampoules should be packed in partitioned boxes. boxes.

LVPLVP

LVP’s which are administered by IV LVP’s which are administered by IV route are commonly called as route are commonly called as IV IV fluids.fluids.

Purposes : Purposes : – Body fluids,Body fluids,– Electrolyte replenisher Electrolyte replenisher

Volume supplied: Volume supplied: 100 to 1000 ml100 to 1000 ml

Precautions / necessities in Precautions / necessities in mfg.:mfg.:

– Free from foreign particles Free from foreign particles – Free from micro organismsFree from micro organisms– Isotonic with body fluidsIsotonic with body fluids– As they are in LVP no bacteriostatic As they are in LVP no bacteriostatic

agents are added agents are added – Free from pyrogens Free from pyrogens

Examples: Examples:

– Dextrose injection IP :Dextrose injection IP : available in 2 , 5 , 10 , 25 & available in 2 , 5 , 10 , 25 & 50 % w/v solution.50 % w/v solution.

– Used for Used for Fluids replenisher,Fluids replenisher, Electrolyte replenisher Electrolyte replenisher

– Sodium chloride & Dextrose injection IP: (DNS)Sodium chloride & Dextrose injection IP: (DNS) Contains Contains

– 0.11 to 0.9 % Sodium chloride 0.11 to 0.9 % Sodium chloride – 2.5 to 5.0 % Dextrose2.5 to 5.0 % Dextrose

– Used for Used for Fluids replenisher,Fluids replenisher, Electrolyte replenisherElectrolyte replenisher Nutrient replenisher Nutrient replenisher

Examples: Examples:

Sodium chloride injection IP: Sodium chloride injection IP: – 0.9 % conc.0.9 % conc.– Also known as normal saline solution Also known as normal saline solution – Used as Used as

Isotonic vehicle Isotonic vehicle Fluids replenisher,Fluids replenisher, Electrolyte replenisherElectrolyte replenisher

Sodium lactate injection IP:Sodium lactate injection IP:– Contains 1.75 to 1.95 % w/v of sodium lactate Contains 1.75 to 1.95 % w/v of sodium lactate – Used as Used as

Fluids replenisher,Fluids replenisher, Electrolyte replenisherElectrolyte replenisher

Examples: Examples:

Mannitol injection IP:Mannitol injection IP:– Contains 5, 10 , 15, 20 % of mannitol Contains 5, 10 , 15, 20 % of mannitol – Used as :Used as :

Diagnostic aidDiagnostic aid Renal function determination Renal function determination As a diuretic As a diuretic

Mannitol & Sodium chloride injection IP:Mannitol & Sodium chloride injection IP:– Contains 5, 10 , 15, 20 % of mannitol & Contains 5, 10 , 15, 20 % of mannitol &

0.45 % of Sodium chloride0.45 % of Sodium chloride – Used as :Used as :

As a diuretic As a diuretic

Examples: Examples:

Other solutions: Other solutions: – Ringer injection IP Ringer injection IP – Ringer lactate solution for injection IPRinger lactate solution for injection IP

Common uses : Common uses : – Used in surgery patients Used in surgery patients – In replacement therapyIn replacement therapy– Providing basic nutrition Providing basic nutrition – For providing TPNFor providing TPN– As a vehicle for other drug subs. As a vehicle for other drug subs.

IV ADMIXTURES IV ADMIXTURES

Definition: Definition: – When two or more sterile products are When two or more sterile products are

added to an IV fluid for their added to an IV fluid for their administration, the resulting administration, the resulting combination is known as IV admixture.combination is known as IV admixture.

– In hospitals, prepared by nurses by In hospitals, prepared by nurses by combining or mixing drugs to the combining or mixing drugs to the transfusion fluids.transfusion fluids.

– The drugs are incorporated in to bottles The drugs are incorporated in to bottles of LV transfusion fluids.of LV transfusion fluids.

Care : Care : – Microbial contamination Microbial contamination – Incompatibility Incompatibility

Physical : change in color Physical : change in color Chemical : hydrolysis, oxidation, reduction Chemical : hydrolysis, oxidation, reduction

etc..etc.. Therapeutic: undesirable antagonistic or Therapeutic: undesirable antagonistic or

synergistic effect synergistic effect

Methods for safe & effective use of IV Methods for safe & effective use of IV admixture:admixture:

– Proper training to nurses & pharmacistProper training to nurses & pharmacist– Instruction regarding labeling Instruction regarding labeling – Information for stability & compatibility Information for stability & compatibility

to the hospital pharmacy dept.to the hospital pharmacy dept.– Information for the formulation skills to Information for the formulation skills to

the pharmacist.the pharmacist.

QC TESTSQC TESTS

Quality control testing and Quality control testing and evaluation of parenterals is evaluation of parenterals is concerned with: concerned with:

I. Incoming raw materials including: I. Incoming raw materials including: routine testing on all drugs, additives, routine testing on all drugs, additives, and packaging materials.and packaging materials.

II. The manufacturing process control II. The manufacturing process control including: including:

1. Daily testing of water for injection.1. Daily testing of water for injection. 2. Confirmation of fill volume and 2. Confirmation of fill volume and

yields of containers.yields of containers. 3. Checking label identity and count.3. Checking label identity and count.

QC TESTSQC TESTS

QC TESTSQC TESTS

III. The Finished product. III. The Finished product.

1. Tests necessary to insure the potency of 1. Tests necessary to insure the potency of the product. the product.

2. Tests for volume in container. 2. Tests for volume in container. 3. Sterility test. 3. Sterility test. 4. Pyrogen test. 4. Pyrogen test. 5. Clarity and particulate analysis. 5. Clarity and particulate analysis. 6. Glass-seal of ampoules “leaker testing”.6. Glass-seal of ampoules “leaker testing”.

QC TESTSQC TESTS Sterility Testing:Sterility Testing: It is a test to estimate the probability of presence of micro-It is a test to estimate the probability of presence of micro- organisms.organisms. Since the test is destructive, it is not possible to examine Since the test is destructive, it is not possible to examine the whole product batch to assure sterility. Adequate the whole product batch to assure sterility. Adequate sampling of the batch must be tested. sampling of the batch must be tested. A sterility test may be done in two ways:A sterility test may be done in two ways: Direct inoculation method:Direct inoculation method: The specified volume of the parenteral product is transferred The specified volume of the parenteral product is transferred

aseptically aseptically to different culture media which expected to to different culture media which expected to support the growth of both support the growth of both aerobic aerobic and and anaerobic anaerobic micro-micro-organisms and media are tested for growth after suitable organisms and media are tested for growth after suitable incubation period. incubation period.

QC TESTSQC TESTS

Thioglycollate mediumThioglycollate medium (to assure anaerobic growth)(to assure anaerobic growth) soybean-casein digest mediumsoybean-casein digest medium ( to assure aerobic growth)( to assure aerobic growth)

QC TESTSQC TESTS

The membrane filtration method:The membrane filtration method: The solution is passed through a membrane having The solution is passed through a membrane having

a pore size sufficiently small to retain bacteria. A a pore size sufficiently small to retain bacteria. A membrane generally suitable for sterility testing membrane generally suitable for sterility testing has a porosity of 0.45 ± 0.02 .The membrane and has a porosity of 0.45 ± 0.02 .The membrane and the retained bacteria on it are transferred to liquid the retained bacteria on it are transferred to liquid media and media are tested for growth after media and media are tested for growth after suitable incubation period.suitable incubation period.

Before conducting the sterility test it is necessary Before conducting the sterility test it is necessary to inactivate any antimicrobial agent which will to inactivate any antimicrobial agent which will interfere with the results of the experiment. This interfere with the results of the experiment. This may be the active drug it self as in case of may be the active drug it self as in case of antibiotic products or it could be a preservative.antibiotic products or it could be a preservative.

QC TESTSQC TESTS

Inactivation procedures:Inactivation procedures:A suitable inactivators may be added to theA suitable inactivators may be added to the

liquid test media to neutralize theliquid test media to neutralize the

antimicrobial substance, e.g. B-Lactamase which hydrolyses antimicrobial substance, e.g. B-Lactamase which hydrolyses

penicillins and cephalosporins.The solution is passed through penicillins and cephalosporins.The solution is passed through

a membrane of pore size small enough to retain bacteria. a membrane of pore size small enough to retain bacteria.

The membrane and the bacteria on it are washed using The membrane and the bacteria on it are washed using salinesaline

solution to remove the last traces of antimicrobial substancesolution to remove the last traces of antimicrobial substance

then inoculated.If solid materials are present they will be then inoculated.If solid materials are present they will be

retained on the filter with the microorganisms, so they retained on the filter with the microorganisms, so they shouldshould

be dissolved in suitable solvent before filtration.be dissolved in suitable solvent before filtration.

QC TESTSQC TESTS

If there is no suitable solvent for the If there is no suitable solvent for the solidsolid

material, the parenteral preparation ismaterial, the parenteral preparation is

diluted to a concentration less than diluted to a concentration less than the MICthe MIC

of the antimicrobial by using a large of the antimicrobial by using a large volumevolume

of medium.of medium.

QC TESTSQC TESTS

Sterility testing of different parenteral Sterility testing of different parenteral products: products:

1. Aqueous solutions are tested by direct 1. Aqueous solutions are tested by direct inoculation or membrane filtration.inoculation or membrane filtration.

2. Solids are dissolved in suitable solvent which has 2. Solids are dissolved in suitable solvent which has no antimicrobial activity then tested by direct no antimicrobial activity then tested by direct inoculation. inoculation.

3. Oily solution of low viscosity are filtered on 3. Oily solution of low viscosity are filtered on membrane and membrane is inoculated. Those of membrane and membrane is inoculated. Those of high viscosity are high viscosity are

diluted first with suitable diluent which has no diluted first with suitable diluent which has no

antimicrobial activity then filtered.antimicrobial activity then filtered.

QC TESTSQC TESTS

4. For oily solution if we used direct 4. For oily solution if we used direct

inoculation method we should inoculation method we should addadd

emulsifying agent emulsifying agent to facilitate mixing ofto facilitate mixing of

oil with the aqueous medium and the oil with the aqueous medium and the

medium should be shaken medium should be shaken every dayevery day

and the and the emulsifying agent should emulsifying agent should

has no antimicrobial activity.has no antimicrobial activity.

QC TESTSQC TESTS

Pyrogen Testing:Pyrogen Testing:

According to the old technique, the USP test is According to the old technique, the USP test is

carried out by injecting in an ear vein of each of 3carried out by injecting in an ear vein of each of 3

rabbits 10 ml of the test solution per kg body rabbits 10 ml of the test solution per kg body

weight, over 10 min. The rabbits should be healthyweight, over 10 min. The rabbits should be healthy

and mature and show temperature change of notand mature and show temperature change of not

more than 1°C from each other and each must not more than 1°C from each other and each must not exceed 39.8°C.exceed 39.8°C.

QC TESTSQC TESTS

The rectal temperature of the rabbits is The rectal temperature of the rabbits is

recorded at 1, 2 and 3 hours subsequent recorded at 1, 2 and 3 hours subsequent to to

injection. The solution is apyrogenic if noinjection. The solution is apyrogenic if no

rabbit shows an individual rise inrabbit shows an individual rise in

temperature of 0.6°C or the sum of thetemperature of 0.6°C or the sum of the

temperature rise of the three rabbits temperature rise of the three rabbits does not exceed 1.4°C.does not exceed 1.4°C.

QC TESTSQC TESTS

If the temperature rise exceeds these If the temperature rise exceeds these limits,limits,

continue the test using another 5 rabbits. continue the test using another 5 rabbits.

The sample is considered acceptable if noThe sample is considered acceptable if no

temperature rise of more than 0.6°C for temperature rise of more than 0.6°C for

each individual rabbit or the sum ofeach individual rabbit or the sum of

temperature rise does not exceed 3.7°C temperature rise does not exceed 3.7°C for the whole 8 rabbits.for the whole 8 rabbits.

QC TESTSQC TESTS

A new method for pyrogen detection isA new method for pyrogen detection is

known as theknown as theLimulus test (LAL).Limulus test (LAL).TheThe

amebocytes, or circulating blood cells, ofamebocytes, or circulating blood cells, of

the horseshoe crab the horseshoe crab (Limulus polyphemus) (Limulus polyphemus)

contain a protein that clots in the presence ofcontain a protein that clots in the presence of

bacterial pyrogens. A test sample is incubated bacterial pyrogens. A test sample is incubated

with amebocytes lysate from the horseshoe crab with amebocytes lysate from the horseshoe crab for 60 min.for 60 min.

QC TESTSQC TESTS

Pyrogenic substance will give A positive Pyrogenic substance will give A positive LAL-LAL-

test characterized by the formation of atest characterized by the formation of a

solid gel that remains intact in the bottomsolid gel that remains intact in the bottom

of the tube on inversion. This methodof the tube on inversion. This method

showed to be more sensitive, more rapidshowed to be more sensitive, more rapid

and easier to perform.and easier to perform.

QC TESTSQC TESTS Weight variation or content uniformity testWeight variation or content uniformity test

This test is intended for sterile solids used for parenteral preparation. This test is intended for sterile solids used for parenteral preparation.

The weight of 10 individual sterile units is noted and the content is The weight of 10 individual sterile units is noted and the content is

removed from them and empty individual sterile unit is weighed intern. removed from them and empty individual sterile unit is weighed intern.

Then net weight is calculated by subtracting empty sterile unit weight Then net weight is calculated by subtracting empty sterile unit weight

form gross weight. The content of active ingredient in each sterile unitform gross weight. The content of active ingredient in each sterile unit

is calculated by performing the assay according to the individual is calculated by performing the assay according to the individual

monographs. The content in 10 sterile units is calculated by performing monographs. The content in 10 sterile units is calculated by performing

the assay. The dose uniformity is met if the amount of active ingredientthe assay. The dose uniformity is met if the amount of active ingredient

is within the range of 85-115.0% of label claim  as determine by theis within the range of 85-115.0% of label claim  as determine by the

content uniformity method or weight variation method..content uniformity method or weight variation method..

QC TESTSQC TESTS

The dose uniformity is also met if the potency value is The dose uniformity is also met if the potency value is 100% in the individual monograph or less of label claim 100% in the individual monograph or less of label claim multiplied by average of limits specified for potency in multiplied by average of limits specified for potency in individual monograph divided by 100 provided that the individual monograph divided by 100 provided that the relative standard deviation in both the cases is equal to or relative standard deviation in both the cases is equal to or less than 6.0%.If one unit is outside the range of 85-less than 6.0%.If one unit is outside the range of 85-115.0%, and none of the sterile unit is outside the range of 115.0%, and none of the sterile unit is outside the range of 75-125.0% and if the relative standard deviation of the 75-125.0% and if the relative standard deviation of the resultant is greater than 6.0% then, the fore mentioned test resultant is greater than 6.0% then, the fore mentioned test is carried for 20 more sterile units. The sterile units meet is carried for 20 more sterile units. The sterile units meet the requirements if not more than one unit is out side the the requirements if not more than one unit is out side the range of 85-115%, no unit is outside the range of 75-range of 85-115%, no unit is outside the range of 75-125.0% and the calculated relative standard deviation is 125.0% and the calculated relative standard deviation is NMT 7.8%NMT 7.8%

QC TESTSQC TESTS

Particulate matter in injectionsParticulate matter in injectionsThe preparations intended for parenteral use should be free form The preparations intended for parenteral use should be free form

particulate matter and should be clear when inspected visually. Twoparticulate matter and should be clear when inspected visually. Two

methods are described by methods are described by USP according to the filled volume of the  according to the filled volume of the

product to be tested. For large volume parenterals (LVP's), a filtrationproduct to be tested. For large volume parenterals (LVP's), a filtration

followed by microscopical examination procedure is used. For smallfollowed by microscopical examination procedure is used. For small

volume parenterals (SVP's)a light obscuration based sensor containingvolume parenterals (SVP's)a light obscuration based sensor containing

electronic liquid-borne particle counter system is used. The USP electronic liquid-borne particle counter system is used. The USP

standards are met if the LVP's under test contain NMT 50 particles perstandards are met if the LVP's under test contain NMT 50 particles per

ml of 10 µm, and NMT 5 particles per ml of 25µm in an effective linearml of 10 µm, and NMT 5 particles per ml of 25µm in an effective linear

dimensional fashion.dimensional fashion.

QC TESTSQC TESTS

The USP standards are met if the The USP standards are met if the SVP's under test contain NMT 10,000 SVP's under test contain NMT 10,000 particles per container of 10 µm, and particles per container of 10 µm, and NMT 1000 particles per container of NMT 1000 particles per container of 25µm in an effective spherical 25µm in an effective spherical diameter.diameter.

QC TESTSQC TESTS