Product Category
a. Topical and non-sterile
respiratory products
b. Oral and rectal products
Product Example
Inhalations, ointments, creams,
gels, paste, topical liquids
Lozenges, capsules,
suppositories
Quantification Specification
Total viable aerobic count- not
more than 102 micro-organisms
(aerobic bacteria plus fungi) per
gram, per millimeter, or per patch
(including the adhesive and
backing layer)
Total viable aerobic count- not
more than 103 bacteria and not
more than 102 fungi per gram or
millimeter
Organisms which must be absent
Transdermal patches: Absence of
Enterobacteriaceae and certain other
gram-negative bacteria, determeined
on 1 patch
Other preparations: not more than 101
Enterobacteriaceae and certain other
gram-negative bacteria per gram or
millimeter
Absence of Pseudomonas
aeuruginosa (1 g, 1 mL, 1 patch)
Absence of Staphylococcus aureus (1
g, 1 mL, 1 patch)
Absence of Escherichia coli (1 g or
mL)
c. Oral products containing
raw materials of natural origin
d. Herbal remedies made
with boiling water
e. Other herbal remedies
Camphor, caffeine, pilocarpine Total viable aerobic count- not
more than 104 bacteria and not
more than 102 fungi per gram or
millimeter
Sage Tea Total viable aerobic count- not
Ginger Tea more than 107 bacteria and not
Thyme Tea more than 105 fungi per gram or
millimeter
Onion Cough Syrup Total viable aerobic count- not
Garlic Honey more than 105 bacteria and not
more than 104 fungi per gram or
millimeter
Absence of Escherichia coli (1 g)
Absence of Staphylococcus aureus(1
g)
Absence of Salmonella (10 g)
Not more than 102Enterobacteriaceae
and certain other gram-negative
bacteria (1 g)
Not more than 102 Escherichia coli
per gram or millimeter
Not more than 103
Enterobacteriaceae and certain other
gram-negative bacteria (1 g)
Absence of Escherichia coli (1 g)
Absence of Salmonella (10 g)
PREPARATIONS FOR ENVIRONMENTAL
MONITORING
ENVIRONMENTAL MONITORING
● Identifies potential routes of contamination
RECOMMENDED ENVIRONMENTAL MONITORING SITES

● Frequently utilized surfaces

○ Storage bins in the work station
○ Shelvings
○ Equipment control panels
● HEPA Filtered Work Station
● Exhaust
● Light switch
ANALYSIS PREPARATION
● Must be performed with a HEPA II Filter Biological Safety Cabinet or a
Laminar flow hood
● Wearing of PPE
● Sample preparation
○ Ensure no tampering of sample
● Media Preparation
○ Nutrient rich general purpose media
○ Neutralizing additives
 METHODS FOR COUNTING
MICROORGANISMS IN A
PHARMACEUTICAL PRODUCT
MEMBRANE FILTRATION METHOD
● Membrane filters- uniform porosity (usually 0.45 µm)
● Sample is passed through the membrane using a FILTER FUNNEL and a
VACUUM SYSTEM
PLATE COUNT METHODS
● Are performed at least in duplicate
● Types:
○ Pour-plate method
○ Surface-spread method
MOST PROBABLE NUMBER METHOD
● Precision and accuracy is less
● Reserved for the enumeration of total aerobic microbial count
Procedures recommended by official pharmacopoeia in
testing specific microorganism
MEDIUM E.coli Salmonella P. aeruginosa S. aureus

LIQUID Enterobacteria Rappaport Enterobacteria Enterobacteria

ENRICHMENT Enrichment Broth Vassiliadis Enrichment Broth Enrichment Broth
Mossel (+) Salmonella Mossel (+) Mossel (-)
Enrichment Broth Rappaport
(+) Vassiliadis
Salmonella
Enrichment Broth
(-)
MEDIUM E.coli Salmonella P. aeruginosa S. aureus

AGAR MEDIUM -Violet Red Bile Xylose Lysine -Violet Red Bile Mannitol Salt Agar
Glucose Agar Deoxycholate Agar Glucose Agar (+)
(++indicative) (++indicative) (++indicative)
-MacConkey Agar -Centrimide Agar
(++indicative) (+)
-Centrimide Agar (-
)
-Mannitol Salt Agar
(-)
MEDIUM E.coli Salmonella P. aeruginosa S. aureus

CONFIRMATION Indole Test Immunoassay strip Oxidase Test Coagulase Test

TEST (+) red colonies, (+) yellow or white
with or without colonies
black centers surrounded by a
yellow zone
STERILITY TEST
Sterility Test
● Is applied to substances or preparations which, according to the
Pharmacopoeia, are required to be sterile. However, a satisfactory result
only indicates that no contaminating microorganism has been found in
the sample examined in the conditions of the test.
Classification of Sterility Test
1) Membrane Filtration

- nominal pore size is not greater than 0.45µm

- diameter of filter used is assumed to be 50mm if a
different diameter is to be used dilutions and washings
should be adjusted
- the filtration apparatus and membrane are sterilized by
appropriate means
Membrane Filtration
● Cellulose Nitrate filter - aqueous,oily, and weakly alcoholic
solutions
● Cellulose Acetate filter - strongly alcoholic solutions
● Specially adapted filters may be needed for certain products (e.g.
antibiotics)
Classification of Sterility Test
2) Direct Inoculation of the culture medium
• Transfer the quantity of the preparation to be examined prescribed in
Table 2
Direct Inoculation of the Culture Medium
• The volume of the product is nmt 10% of the volume of the medium,
unless otherwise prescribed
• If product has antimicrobial activity carry out test after neutralizing
• If a large volume of product is to be used a concentrated culture
medium medium should be prepared
PYROGEN TESTING
Pyrogen

● fever-inducing substances
● harmful or even fatal if administered to humans above certain
concentrations
LAL TEST (limulus amebocyte lysate testing)
● Bacterial endotoxin testing
● in vitro assay
● detect the presence and concentration of bacterial endotoxins in drugs and biological products
Endotoxins
● type of pyrogen
● lipopolysaccharides present in the cell walls of gram-negative bacteria
● react with limulus amebocyte lysate

LAL Test

● Samples are mixed with the LAL reagent in a

96 well plate and a plate reader measure the
color change over time
● rate of that color change is proportional to the
Rabbit Pyrogen Test
● in vivo test
● detect pyrogens qualitatively
● Rabbits - similar pyrogen tolerance to humans
● Observable change in the rabbit’s body temperature - determination of the presence of pyrogens
● can detect non-bacterial endotoxin pyrogens as well as bacterial endotoxins