Asst. Prof. Dr. Mehmet İLKTAÇ

Asst. Prof. Dr. Mehmet İLKTAÇ

PHAR457

Lecture 2

Contain particular number of microorganisms as a result of their nature.

NON-STERILE PHARMACEUTICALS

Quantities higher than acceptable threshold and/or presence of objectionable microorganisms

Spoilage of the pharmaceutical

Health threat to consumers

1. Product recalls 2. Production shutdown

3. Losses in labor and manufacturing 4. Financial loss

CONSEQUENCES OF CONTAMINATION

Although non-sterile, NON-STERILE PHARMACEUTICALS

Quantity + Types of m.o

Safety of the product and efficacy of manufacturing

process

Microbiological testing of non-sterile pharmaceuticals = Microbial limit tests

determines

‘’OBJECTIONABLE MICROORGANISMS’’

‘’Microorganisms which might cause serious health threat to consumers, might cause drug spoilage or

indicate the presence of other pathogenic bacteria’’

a. Total Aerobic Microbial Count

b.Total Combined Yeast and Mould Count

QUALITY ACCEPTANCE CRITERIA OF NON-STERILE PHARMACEUTICALS

1. Quantity of microorganisms (Quantitative tests)

2. Absence of specific indicator microorganisms (Qualitative tests)

Salmonella Escherichia coli Pseudomonas aeruginosa

Staphylococcus aureus Candida albicans Bile tolerant Gram negative bacteria Clostridia

INDICATOR OBJECTIONABLE MICROORGANISMS (SPECIFIED MICROORGANISMS)

Skin and GIS infections and toxic shock

syndrome

Opportunistic infections

Gastrointestinal system disorder (AGE)

Fecal coliform and some strains cause GIS diseases

Coliform and Opportunistic pathogen

Wound, intestinal and neurological infection

Vaginitis

E. coli

E. coli Klebsiella Enterobacter Citrobacter

Coliform bacteria: Rod shaped Gram negative Non-spore forming Ferment lactose with the production of acid and gas

when incubated at 35–37°C for 24 hours.

Fecal Coliform bacteria: Rod shaped Gram negative

Non-spore forming Ferment lactose with the production of acid and gas

when incubated at 44°C for 24 hours.

INDICATOR OF FECAL CONTAMINATION

BETTER INDICATOR FOR FECAL

CONTAMINATION

COLIFORMS: INDICATOR OF FECAL CONTAMINATION OF PRODUCTS

Is the indicative of failed GMP in industry.

THE USE OF SEVEN INDICATOR BACTERIA DOES NOT MEAN THAT THE PRESENCE OF OTHER BACTERIA MIGHT NOT BE A PROBLEM DURING QUALITY EVALUATION.

ROUTE OF APPLICATION AND DELIVERY SYSTEM OF DRUG

INDICATION FOR USE (INTENDED CONSUMER)

CHEMICAL COMPOSITION OF PRODUCT PRODUCTION PROCESS (FILTRATION, HEATING, DRYING...)

WILL DETERMINE IF THERE IS A RISK

INVOLVED WHEN OTHER

MICROORGANISMS PRESENT

BACTERIAL INDICATORS DO NOT INCLUDE ALL THE OBJECTIONABLE BACTERIA PRESENT IN THE

ENVİRONMENT

Processes are NOT controlled Water, air and environmental monitoring are NOT performed

NON-STERILE PHARMACEUTICALS Manufactured under aseptic conditions

BUT As regularly as

sterile manufacturing

GMP, aseptic thecniques Environmental control Personnel and equipment sanitation

Necessary to control the

presence and the number of

microorganisms

Limit testing is important for quality control analysis of non sterile products.

Microcrystalline Cellulose Capsule Shells Magnesium Stearate Xanthan Gum Sodium Starch Glycolate Lactose Monohydrate Lactose Anhydrous Talc Powdered Cellulose Cellulose Acetate Povidone

Copovidone Hydroxyethyl Cellulose Hydroxypropylcellulose Sugar Mannitol NF Corn Starch Glucose Croscarmellose Sodium NF Cotton Coil Sugar Spheres Opadry

RAW MATERIALS THAT MAY NEED MICROBIAL LIMIT TESTING

Metered-dose and dry powder inhalants Nasal sprays Otics Vaginal suppositories Topicals Rectal suppositories Oral liquids (aqueous) Liquid-filled capsules Oral tablets and powder-filled capsules

CATEGORIES OF NONSTERILE PHARMACEUTICAL PRODUCTS

Decreasing risk of contamination

Antimicrobial activity

Antimicrobial preservative

Exipient microbial content

Water activity

Manufacturing processes

WHAT ARE THE SOURCES OF MICROBIAL

CONTAMINATION OF NON-STERILE

PRODUCTS

SOURCES OF CONTAMINATION 1.) Pharmaceutical ingredients (API, exipient, raw mat) Animal and plant origined raw materials naturally contain microorganisms.

Raw materials should be tested prior to use in manufacturing.

2.) Water

Water system must be validated.

Water system and water should be monitored regularly to reduce the microbial load. Water system must be sanitized regularly to prevent microorganism colonisation and biofilm formation.

Heat treatment Chemical treatment

UV treatment Filtration

SOURCES OF CONTAMINATION 3.) Equipment and building areas

Building areas and equipment should be sanitized properly.

Environmental monitoring is important in order to reduce microbial contamination.

SOURCES OF CONTAMINATION 4.) Manufacturing Personnel and noncompliance with GMP

Face masks, gloves, laboratory coat, hair coverers and safety glasses should be worn

Training of manufacturing and lab. personnel

Normal flora in oral cavity, skin, nasopharynx etc.

SOURCES OF CONTAMINATION 5.) Air quality

Air is generally contaminated with moulds and bacterial spores.

Air quality control of both living and non living particulates should be carried out.

4 pt

4 pt

4 pt 4 pt

4 pt

4 pt

4 pt

4 pt 1

2

3

4

Less stringent GMP compliance

Lack of functional microbial limit testing (raw materials and finished products) program

Lack of equipment sanitization and environmental sampling

Lack of properly trained personnel

NON-STERILE PHARMACEUTICAL CONTAMINATION

INCREASES THE RISK OF

CONTAMINATION, ALLOWS

OBJECTIONABLE MICROORGANISMS TO CONTAMINATE

PRODUCTS AND INCREASES THE

LOAD OF MICROBIAL

CONTAMINATION

MOST COMMON CONTAMINANTS Gram (-) rods are the most common contaminants.

Pseudomonas Burkholderia cepacia Ralstonia pickettii

Molds and yeasts are also among common contaminants.

Gram (+) bacteria

Staphylococcus

Streptococcus

Bacillus

Clostridium

Personnel origined

Lack of process

control of environment

Lack of personnel hygiene

Water and raw material origined

II.) Spoilage of pharmaceuticals

I.) Health threat to consumers

IMPORTANCE OF USP BACTERIAL INDICATOR

Some strains of E. coli

S. aureus

Salmonella GIS disease in healthy patients

GIS disease in healthy patients

Skin and GIS disease in healthy patients

Pseudomonas, Enterobacteriaceae

and others

Opportunistic infection

I.) Health threat to consumers

Opportunistic: Infections not seen in healthy patients but in :

Immuncompromised patients

Patients with underlying disease

Newborn infants

Elderly people

FOR THESE PATIENTS,

LIMITS OF THE PRODUCTS MUST BE

LOWER THAN FOR PEOPLE

WITH FUNCTIONAL

IMMUNE SYSTEM

IMPORTANCE OF USP BACTERIAL INDICATOR

Microbial limit tests of raw material and finished products determine the

MICROBIOLOGICAL QUALITY of non-sterile pharmaceutical products.

MICROBIAL LIMIT TESTING

DETERMINES TOTAL THE NUMBERS OF BACTERIA, YEAST AND MOULDS PRESENT IN THE SAMPLE

According to USP, EP and JP, microbial limit testing is divided into 2 different tests:

MICROBIAL LIMIT TEST METHODS

DETERMINES THE PRESENCE OF SPECIFIC INDICATOR (OBJECTIONABLE) MICROORGANISMS

Microbial Bioburden: Total number and type of microorganisms present in a pharmaceutical.

Quantitative tests

Qualitative tests

MICROBIAL LIMIT TESTS (MLT)

Quantitative tests

Qualitative tests

MICROBIAL LIMIT TESTS (MLT)

Three pharmacopoeia MICROBIAL LIMIT TEST METHODS

Prior to 2006, used test methods that were similar but widely variable in practice and acceptance criteria.

They harmonized their testing methods, specifications and acceptance criteria for non-sterile pharmaceutical products.

USP EP JP

I. QUANTITATIVE TESTS (ENUMERATION TESTS)


The ability of the test to detect and queantify bacteria, yeasts and moulds in the presence of the product to be tested must be established.

QUANTITATIVE TESTS

SUITABILITY OF QUANTITATIVE TEST

SUITABILITY OF QUANTITATIVE TEST Preparation of test strains

Staphylococcus aureus Pseudomonas aeruginosa Bacillus subtilis Candida albicans Aspergillus brasiliensis

Test strains for

quantitative validation

S. aureus, P. aeruginosa and Bacillus subtilis are grown on Soybean Casein Digest broth (SCDB) at 37 °C for 18-24 hours.

C. albicans is grown on Sabouraud Dextrose broth at 20-25 °C for 2-3 days and A. brasiliensis is grown on Sabouraud Dextrose agar at 20-25 °C for 5-7 day.

SUITABILITY OF QUANTITATIVE TEST Preparation of test strains

Buffered sodium chloride peptone (BSCP) solution, phosphate buffer solution (PBS) or SCDB is used as diluent to make test suspension.

Use suspensions within 2 hours (stored at room temperature) or 24 hours if stored at 2-8 °C.

Need to do 10 fold serial dilutions because maximum 100 cfu microorganism inoculum is needed to be obtained in the further steps.

Negative control is performed with only diluent solution having no microorganism. Should be no growth in the final analysis.

All media should be tested for growth promotion.

Negative control

Growth promotion of the media

Growth obtained must not differ by a factor greater than 2 from the calculated value for standardized inoculum.

Preparation of test strains

Preparation of Microorganisms and Growth Promotion Tests

SUITABILITY OF QUANTITATIVE TEST Preparation of Drug Samples To be Tested Water soluble products: Dilute the 10 ml or gr of the product 1:10 in BSCP, PBS or SCDB.

Non fatty products Insoluble In Water: Add polysorbate 80 (1g/lt) after diluting 10 ml or gr of the product 1/10 in BSC, PBS or SCDB.

Fatty Products: Dissolve in isopropyl myristate or mix with minimum quantity of sterile preheated (40 °C) polysorbate 80. Dilute 1/10 with diluent.

Transdermal patches: Remove protective cover sheets of 10 patches, cover adhesive surface with sterile gauze and transfer to 500 ml diluent. Shake 30 minutes.

SUITABILITY OF QUANTITATIVE TEST Inoculation and dilution

Sufficient volume of diluted microbial suspension is added to the diluted product and to the POSITIVE CONTROL with no product (positive control will be compared with the result of product to asses the suitability of method ).

The final inoculum of microorganism should not be more than 100 cfu/ml.

The volume of suspension of microorganism should not be more than 1% of that of diluted product.

Diluted microorganism

Diluted product

INOCULATION AND DILUTION

RECOVERY AND ENUMERATION OF MICROORGANISMS IN THE PRESENCE OF PRODUCT

1. PLATE COUNT METHODS 2. MEMBRANE FILTRATION METHOD

3. MOST PROBABLE NUMBER (MPN)METHOD

The least accurate method for microbial counts

Which one to choose ? Required limit of microorganisms

Nature of the product

SUITABILITY OF QUANTITATIVE TEST

depends

RECOVERY AND ENUMERATION OF MICROORGANISMS 1. PLATE COUNT METHODS

Perform plate count methods at least in duplicate for each strain and use the mean (avarage) count.

A. POUR PLATE METHOD

1 ml of sample (diluted product + diluted m.o + neutralization agent)

15-20 ml of Soybean-Casein Digest agar or Saboraud Dextrose agar at 45 °C

+

Add to petri dish, stir and incubate as stated in prep. of test strains

Diluted product

and bacteria

Inoculate empty plate

Add melted SCDA (15-20 ml)

Swirl to mix

Colonies grown on solidified media

POUR PLATE METHOD

1 ml

STREAK OF DIFFERENT DILUTED SERIES OF MICROORGANISMS

CHOOSE THE LOWEST DILUTION WHICH HAS LESS THAN 100 CFU

RECOVERY AND ENUMERATION OF MICROORGANISMS 1. PLATE COUNT METHODS

B. SPREAD PLATE METHOD

Add 15-20 ml Soybean-Casein Digest agar or Sabouraud Dextrose Agar into a petri dish, allow it to solididy.

Spread at least 0.1 ml of the sample (diluted product + m.o + neutralization agent) over the surface.

Incubate as stated in prep. of test strains.

Count cfu of the duplicates, take the average and calculate the number of cfu in original inoculum

SPREAD PLATE METHOD

Diluted product and bacteria

0.1 ml

Inoculate petri dish containing solid medium

Spread inoculum over surface evenly

Colonies grow on surface of medium

RECOVERY AND ENUMERATION OF MICROORGANISMS 2. MEMBRANE FILTRATION METHOD

Membrane filters with 0.45µm pore size

Sample (representing 1 gr of the sample; generally 10 ml) is transferred into membrane filter and filtrated.

For total aerobic microbial count (TAMC), place the filter onto Soybean-Casein Digest agar.

For total combined yeasts and moulds count (TYMC), put the filter on to Sabouraud Dextrose agar.

RECOVERY AND ENUMERATION OF MICROORGANISMS 3. MOST PROBABLE NUMBER (MPN) METHOD

The accuracy of MPN method is less than that of plate count methods and membrane filtration.

Not suitable especially for yeasts and moulds.

The MPN is particularly useful for low concentrations of organisms

Prepare 10-1, 10-2 and 10-3 dilutions of sample METHOD

From each dilution, add 1 ml to to each of three tubes containing 9 ml Soybean-Casein Digest Broth.

Incubate and analyse according to the table.

MPN TABLE

RESULTS AND ANALYSIS SUITABILITY OF QUANTITATIVE TEST

1. Plate Count Methods

2. Membrane Filtration

Mean count of test organism with product must not differ by a factor greater

than 2 from the value of POSITIVE CONTROL

(contains no product)

To verify suitability of,

3. MPN The calculated value from inoculum must be within 95% confidence limits of

results obtained with CONTROL.

> 50% and < 200%

If the number of microorganisms recovered from the diluted product (after the end of the test) is less than

a factor greater than 2 compared to the number of microorganism recovered from positive control

preparation

SUITABILITY OF QUANTITATIVE TEST Neutralization and Removal of Antimicrobial Activity

THE PRODUCT HAS ANTIMICROBIAL ACTIVITY

HAVE TO NEUTRALIZE/REMOVE IT

SUITABILITY OF QUANTITATIVE TEST Neutralization and Removal of Antimicrobial Activity

Increase the volume of diluent (eg. dilute 1/100) Membrane Filtration

Add neutralizing agent into diluent

Agents used for neutralization and as surface active substance SHOULD HAVE NO TOXICITY ON MICROORGANISMS

Lecitin Polysorbate

Glycine

Thioglycollate

Thiosulphate

Ca+2 Mg+2

Sodium bisulphite

If all neuralization methods fail to neutralize the product, failure to isolate the inoculated organism is due to microbicidal activity of the product.

Neutralization and Removal of Antimicrobial Activity

The product cannot be contaminated with that microorganism species.

However, other microorganisms not included among the test strains can contaminate this product. Therefore, microbial limit testing must be performed in case of the contamination of other microorganisms.

QUANTITATIVE TESTING OF PRODUCTS

QUANTITATIVE TESTING OF PRODUCTS After methods are validated (suitability testing), non-sterile products can be tested by quantitave tests routinely.

AMOUNT OF PRODUCT USED FOR TEST Generally 10 g or ml

Aerosol dosage forms: 10 containers

Transdermal patches: 10 patches

Tablet, capsule and injection: 10 units

Drug substances that is less than 1000 ml or 1000 gr: 1% of the batch

QUANTITATIVE TESTING OF PRODUCTS AMOUNT OF PRODUCT USED FOR TEST

Distribution of microorganisms in a batch is not homogenous.

They can clump together only in some containers in the same batch.

The sample of volume of 10 ml or gr MUST BE A COMPOSITE OF 10 different containers.

QUANTITATIVE TESTING OF PRODUCTS EXAMINATION OF THE PRODUCT

MEMBRANE FILTRATION Transfer the appropriate amount (representing one gram of sample) of prepared product to each of two 0.45 µm membrane filters. 50 ml of prepared product for transdermal patches are filtered.

After filtration

Total aerobic microbial count

Total yeast mold count

Transfer filter onto SCDA

Transfer filter onto SDA

Incubate 3-5 days at 37 °C

Incubate 5-7 days at 25 °C

MOST APPROPRIATE METHOD FOR PRODUCTS CONTAINING ANTIMICROBIAL SUBSTANCES


POUR PLATE METHOD Ten fold serial dilution of sample is prepared. 10-1, 10-2, 10-3

For each dilution 2 SCDA (for TAMC) and 2 SDA (TYMC) are used.

Incubate media for 3-5 days at 35-37 C for bacteria and 5-7 days at 20-25 C for fungi.

After incubation, select the dilution showing the highest number of colonies less than 250 for TAMC and 50 for TYMC.

Take the arithmetic mean of counts of 2 media.

ANALYSIS OF PRODUCT WITH POUR PLATE METHOD


SPREAD PLATE METHOD

Follow the the same procedures as in Pour plate method.

The only difference is that the sample and the media are not poured together. First media is poured into petri dish and after it has solidified 0,1 ml of the product is spread over the media.


MOST PROBABLE NUMBER

Prepare dilutions of 10-1, 10-2 and 10-3.

Transfer 1 ml each of dilution to 3 tubes containing 9 ml SCDB.

Record the number of tubes that are positive for growth for each dilution.

Determine the most probable number of organism according to table.

Incubate at 37 °C for 5 days.

QUANTITATIVE TESTING OF PRODUCTS INTERPRETATION OF RESULTS

TAMC Number of cfu found on SCDA.

TYMC Number of cfu found on SDA.

Acceptance criteria for nonsterile pharmaceutical products based upon the total aerobic microbial count (TAMC), the total combined yeasts and molds count (TYMC) and the absence of specified microorganisms.

Total Aerobic Microbial Count

Total Combined Yeast and Mould Count


CALCULATION OF CFU/ml

Dil Fac=10/ml per plate

101 cfu : maximum acceptable count = 20; 102 cfu: maximum acceptable count = 200; 103 cfu: maximum acceptable count = 2000 ....

When an acceptance criterion for microbiological quality is analysed, it is interpreted as follows:


If the acceptance criterion is;

RECOMMENDED ACCEPTANCE CRITERIA FOR MICROBIOLOGICAL QUALITY OF PHARMACEUTİCAL PREPARATIONS (TOTAL COUNTS)

102

102

102

102

102

102

102

103

103

102

102

101

101

101

101

101

101

101

101

102

102

101

RECOMMENDED ACCEPTANCE CRITERIA FOR MICROBIOLOGICAL QUALITY OF RAW MATERIALS FOR

PHARMACEUTICAL USE (TOTAL COUNTS)

103 102

Psychrophilic, thermophilic, basophilic and anaerobic bacteria and microorganisms which require specific ingredients for growth may give a NEGATIVE RESULT, even if they exist in a significant number.

Quantitative testing determines only mesophilic bacteria and fungi which grow under aerobic conditions.

QUANTITATIVE TESTS

II. QUALITATIVE TESTS


QUALITATIVE: PRESENCE OR ABSENCE

The presence of certain microorganisms in nonsterile preparations may have the potential:

To reduce or inactivate the therapeutic activity of the product To adversely affect the health of the patient.

To be indicative of pathogen bacteria that may

present

These certain risky microorganisms are refered to as ‘’OBJECTIONABLE MICROORGANISMS’’.

In qualitative quality control tests of nonsterile products, the presence/absence of some indicator ‘’objectionable microorganisms’’ are investigated.

Salmonella Escherichia coli Pseudomonas aeruginosa Staphylococcus aureus

Bile tolerant Gram negative bacteria Clostridium

Candida albicans

INDICATOR ORGANISMS WHICH USP/EP/JP PROVIDE PROCEDURES AND TEST CONDITIONS

FOR DETERMINING WHETHER THE

PRODUCT MEETS THE ACCEPTANCE

CRITERIA OF QUALITY

INDICATOR (SPECIFIED) MICROORGANISMS

Salmonella Escherichia coli Pseudomonas aeruginosa

Staphylococcus aureus

Candida albicans Bile tolerant Gram negative bacteria Clostridium

Skin and GIS infections and toxic shock

syndrome

Opportunistic infections

Gastrointestinal system disorder (AGE)

Fecal coliform and some strains cause GIS diseases

Coliform and Opportunistic pathogen


Wound, intestinal and neurological infection

Vaginal infections


Indicators donot include all the objectionable microorganisms found in the environment.

Other types of bacteria can also contaminate and be responsible of product recalls.

Burkholderia cepacia Acinetobacter Pseudomonas putida Bacillus cereus ......

Objectionable screening testing MUST NOT BE LIMITED TO THESE INDICATORS. MUST INCLUDE OTHER PATHOGENS OR OPPORTUNISTIC MICROORGANİSMS THAT MAY GENERATE HEALTH THREAT TO CONSUMERS OR MAY SPOIL THE PRODUCT FORMULA.


Some of the frequently isolated microorganisms from a production facility can be added as an indicator.

Pharmacopoeias donot include all objectionable microorganisms. The significance of other microorganisms recovered should be evaluated in terms of the following:

The chemical composition of the product: Does the product support growth? Does it have adequate antimicrobial preservation? The intended consumer: Risk may differ for neonates, infants, the debilitated. Use of immunosuppressive agents, corticosteroids. The presence of disease, wounds, organ damage. Production process: Heating, blending, filtration, drying, shearing ....

Route of administration: Hazard varies according to the route of administration (oral,ocular, aural, topical, injection, inhalation, rectal, vaginal ....).

Number of microorganism on plate: If high, then identify

II. QUALITATIVE TESTS Qualitative tests use selective

media for the recovery indicator microorganisms.

Selective media cannot detect subletally injured microorganisms.

As subletally injured microorganisms are relevant for

the quality of the product, A RESUSCİTATİON

(PREINCUBATION) STEP MUST BE INCLUDED.

II. QUALITATIVE TESTS TESTING OF PRODUCTS

Preparation of sample (product) is carried out as described in quantitative methods.

If the product has antimicrobial activity it has to be removed or neutralized by a neutralization agent which has no toxicity on microorganisms.

Surface active substances must also have no antimicrobial activity against microorganisms.


The product is diluted 1:10 using not less than 10 gr or 10 ml of product (as in enumeration method).

SALMONELLA

Inoculate Soybean Casein Digest Broth with 10 ml of the sample solution.

1.) Sample preparation and Preincubation:

Incubate for 18-24 hours at 35-37 C.


SALMONELLA 2.) Selection and subculture: Transfer 0.1 ml of SCDB to 10 ml RAPAPORT-VASILIADIS SALMONELLA enrichment broth and incubate 35 C for 18-24 hours.

RAPAPORT-VASILIADIS IS A SELECTIVE ENRICHMENT BROTH FOR SALMONELLA

From Rappaport, subculture on Xylose Lysine Deoxycholate agar. Incubate at 35-37 °C for 18 to 48 hours.

TESTING OF PRODUCTS SALMONELLA


There is an enrichment step for Salmonella spp. using Rappaport-Vasiliadis broth.

TESTING OF PRODUCTS SALMONELLA

3.) Interpretation:


Red colonies with black centers are indicative of Salmonella.

Confirm with biochemical tests even if the colony morphology is different than expected.


SALMONELLA Rappaport Vasiliadis

Xylose Lysine Deoxycholate

SCDB RVSEM

II. QUALITATIVE TESTS - SALMONELLA

XLDA


OTHER STRAINS APART FROM SALMONELLA

1.) Sample preparation and Preincubation:

For C. albicans Sabouraud Dextrose broth is used instead of SCDB in preincubation for 24-48 hours.

For the other strains apart from Salmonella, at least 1 gr or 1 ml of the product is diluted 1:10 except for Clostridium in which 2 gr or 2 ml of the product is diluted.

For Clostridium, diluted (1:10) product is divided into 2 portions of at least 10 ml. One portion is heated at 80 C; the other is not.

2.) Selection and subculture:


OTHER STRAINS APART FROM SALMONELLA

Do not forget to incubate Clostridium under ANAEROBIC conditions.


E. coli MacConkey Broth

MacConkey Agar


Pseudomonas aeruginosa

Cetrimide agar


Staphylococcus aureus

Mannitol salt agar


Candida albicans Sabouraud Dextrose broth


Clostridium

MEDIA USED IN SELECTION AND SUBCULTURE FOR MICROORGANISMS

Acceptance Criteria for NONSTERILE Pharmaceutical Preparations and Substances for

Pharmaceutical Use

1.) Total aerobic microbial count (TAMC) 2.) The total combined yeasts and molds count (TYMC) 3.) Absence of specified microorganism

TESTING FREQUENCY There is no requirement for every batch of non-sterile medicine to be tested for microbial quality prior to release.

Periodic testing or 'skip-lot' testing can be performed.

The frequency of testing should be determined based on:

The bioburden history of the medicine The nature of product The manufacturing process for the medicine The controls that are inherent in GMP

A BIOBURDEN HISTORY FOR THE MEDICINE The first 5 to 10 batches of a new medicine should be tested for microbial quality prior to release.

If test results for these batches are satisfactory, then testing could be performed periodically, rather than on every batch.

NATURE OF PRODUCTS Some products have antimicrobial activity

Require little monitoring

Products which have higher risk of contamination require more frequent testing than products with low risks.

TESTING FREQUENCY

Metered-dose and dry powder inhalants Nasal sprays Otics Vaginal suppositories Topicals Rectal suppositories Oral liquids (aqueous) Liquid-filled capsules Oral tablets and powder-filled capsules

CATEGORIES OF NONSTERILE PHARMACEUTICAL PRODUCTS

Decreasing risk of contamination

Antimicrobial activity

Antimicrobial preservative

Exipient microbial content

Water activity

Manufacturing processes

Some processes include filtration, drying, heating processes which have detrimental effect on microorgansims.

TESTING FREQUENCY MANUFACTURING PROCESS FOR THE MEDICINE

Such products are tested less frequently than those whose manufacturing donot include such processes.

CONTROLS IN GMP

The more strictly adherence to GMP, the less the risk of contamination and the less frequent the testing.

Asst. Prof. Dr. Mehmet İLKTAÇ

Documents

Transcript of Asst. Prof. Dr. Mehmet İLKTAÇ