Mikrobiologisk kvalitet av legemidler

Microbiological Quality of Non-sterile Pharmaceutical Preparations and Substances for Pharmaceutical Use
(Ph. Eur. 5.1.4.)

 

 

The presence of certain micro-organisms in non-sterile preparations may have the potential to reduce or even inactivate the therapeutic activity of the product and has a potential to adversely affect the health of the patient. Manufacturers therefore have to ensure a low bioburden of finished dosage forms by implementing current guidelines on Good Manufacturing Practice during the manufacture, storage and distribution of pharmaceutical preparations.

 

Microbial examination of non-sterile products is performed according to the methods given in general chapters 2.6.12 and 2.6.13. Acceptance criteria for non-sterile pharmaceutical products based upon the total aerobic microbial count (TAMC) and the total combined yeasts/moulds count (TYMC) are given in Tables 5.1.4.-1 and 5.1.4.-2. Acceptance criteria are based on individual results or on the average of replicate counts when replicate counts are performed (e.g. direct plating methods).

 

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

-    10¹ CFU: maximum acceptable count = 20;

-    10² CFU: maximum acceptable count = 200;

-    10³ CFU: maximum acceptable count = 2000, and so forth.

 

Table 5.1.4.-1 includes a list of specified micro-organisms for which acceptance criteria are set. The list is not necessarily exhaustive and for a given preparation it may be necessary to test for other micro-organisms depending on the nature of the starting materials and the manufacturing process.

 

If it has been shown that none of the prescribed tests will allow valid enumeration of micro-organisms at the level prescribed, a validated method with a limit of detection as close as possible to the intended acceptance criterion is used.

 

Table 5.1.4.-1. – Acceptance criteria for microbiological quality of non-sterile dosage forms

 

Route of administration	TAMC (CFU/g or CFU/ml)	TYMC (CFU/g or CFU/ml)	Specified microorganisms
Non-aqueous preparations for oral use	103	102	Absence of Escherichia coli   (1 g or 1 ml)
Aqueous preparations for oral use	102	101	Absence of Escherichia coli   (1 g or 1 ml)
Rectal use	103	102
Oromucosal use Gingival use Cutaneous use Nasal use Auricular use	102	101	Absence of Staphylococcus aureus    (1 g or 1 ml) Absence of Pseudomonas aeruginosa   (1 g or 1 ml)
Vaginal use	102	101	Absence of Pseudomonas aeruginosa   (1 g or 1 ml) Absence of Staphylococcus aureus   (1 g or 1 ml) Absence of Candida albicans   (1 g or 1 ml)
Transdermal patches (limits for one patch including adhesive layer and backing)	102	101	Absence of Staphylococcus aureus (1 patch) Absence of Pseudomonas aeruginosa (1 patch)
Inhalation use (special requirements apply to liquid preparations for nebulisation)	102	101	Absence of Staphylococcus aureus   (1 g or 1 ml) Absence of Pseudomonas aeruginosa   (1 g or 1 ml) Absence of bile-tolerant gram-negative bacteria   (1 g or 1 ml)
Special Ph. Eur. provision for oral dosage forms containing raw materials of natural (animal, vegetal or mineral) origin for which antimicrobial pretreatment is not feasible and for which the competent authority accepts TAMC of the raw material exceeding 103 CFU per gram or per millilitre	104	102	Not more than 102 CFU of bile-tolerant gram-negative bacteria   (1 g or 1 ml) Absence of Salmonella   (10 g or 10 ml) Absence of Escherichia coli   (1 g or 1 ml) Absence of Staphylococcus aureus   (1 g or 1 ml)
Special Ph. Eur. provision for herbal medicinal products consisting solely of one or more herbal drugs (whole, reduced or powdered): -herbal medicinal products to which boiling water is added before use (1)   -herbal medicinal products to which boiling water is not added before use (2)	107     105	105     104	(1) Not more than 102 CFU of Escherichia coli (see Appendix) (1 g or 1 ml) (2) Not more than 103 CFU of bile-tolerant gram-negative bacteria   (1 g or 1 ml) Absence of Escherichia coli    (1 g or 1 ml) Absence of Salmonella   (10 g or 10 ml)

 

 

Table 5.1.4.-2. – Acceptance criteria for microbiological quality of non-sterile substances for pharmaceutical use

 

	TAMC (CFU/g or CFU/ml)	TYMC (CFU/g or CFU/ml)
Substances for pharmaceutical use	103	102

 

In addition to the microorganisms listed in Table 5.1.4.-1, the significance of other micro-organisms recovered is evaluated in terms of:

-    use of the product: hazard varies according to the route of administration (eye, nose, respiratory tract);

-    nature of the product: its ability to support growth, the presence of adequate antimicrobial preservation;

-    method of application;

-    intended recipient: risk may differ for neonates, infants, the debilitated;

-    use of immunosuppressive agents, corticosteroids;

-    presence of disease, wounds, organ damage.

 

Where warranted, a risk-based assessment of the relevant factors is conducted by personnel with specialised training in microbiology and the interpretation of microbiological data. For raw materials, the assessment takes account of processing to which the product is subjected, the current technology of testing and the availability of materials of the desired quality.

 

Appendix: special Ph. Eur. provision for herbal medicinal products consisting solely of one or more herbal drugs (whole, reduced or powdered): quantificative test for E.coli

 

Use the following protocol.

 

Sample preparation and pre-incubation. Prepare a sample using a 10-fold dilution of not less than 1 g of the product to be examined as described in general chapter 2.6.12, and use the quantities corresponding respectively to 0.1 g, 0.01 g and 0.001 g (or 0.1 ml, 0.01 ml and 0.001 ml) to inoculate a suitable amount (determined as described under 3-4 of general chapter 2.6.13) of casein soya bean digest broth, mix and incubate at 30-35 °C for 18-24 h.

 

Selection and subculture. Shake the container, transfer 1 ml of casein soya bean digest broth to 100 ml of MacConkey broth and incubate at 42-44 °C for 24-48 h. Subculture on a plate of MacConkey agar at 30-35 °C for 18-72 h.

 

Interpretation. Growth of colonies indicates the possible presence of E. coli. This is confirmed by identification tests.

 

Note the smallest quantity of the product that gives a positive result and the largest quantity that gives a negative result.

 

Determine from the following table the probable number of bacteria.

 

Results for each quantity of product			Probable number of bacteria per gram or millilitre of product
0.1 g or 0.1 ml	0.01 g or 0.01 ml	0.001 g or 0.001 ml
+	+	+	> 103
+	+	-	< 103 and > 102
+	-	-	< 102 and > 10
-	-	-	< 10

Sist gjennomgått: 04.01.2010
Første gang publisert: 12.01.2009