1. Introduction
Cleaning validation is study done to find the
effectiveness and reliability of cleaning pharmaceutical production equipment.
People in the pharmaceutical industry use equipment validation and cleaning
procedures mainly to prevent cross-contamination that makes these practices
crucial. In general, cross-contamination usually happens when an active
ingredient from one product is transferred to other product through the
instruments improper cleaning that can pose real risks to consumers and second
type is contamination by foreign materials, which could be bacteria or fungi
.Poor maintenance or storage conditions may let microbes flourish in processing
equipment and becomes a serious issue
Scope of study
To perform cleaning validation study on equipments used for formulation of products ( Clobetasol 0.05% cream )
3.
Methods
3.0:
Study Design
The study was designed as experimental study and
conducted between January-June 2025.The study was conducted on Clobetasol 0.05%
cream manufacturing equipments in class D manufacturing facility in pharmaceutical
Industry. Sampling method was selected as Swabbing technique, to study the
microbial load on equipments included the study. Total aerobic Microbial Count
(TAMC) 100 cfu/ 100 cm 2 and Total Yeast and Mould Count 10cfu/ 100
cm 2 was set as pass limit during cleaning
Validation study..
a.To perform cleaning validation study on equipments used for formulation of products(clobetasol cream 0.05%)
Test conditions of cleaning validation was designed
under static condition .Temperature less (≤ 25
◦C),
and Humidity (≤ 60%) was maintained
in sampling areas during study periods. Swabbing technique was used for
sampling and membrane filter test method was used for detection of microbial load
on equipments included in the study.
3.2:
Materials Used under study
Materials and
Manufacturer
1
Buffered peptone water: Hi Media
2
Soyabean Casein Digest Agar (SCDA): Hi
Media
3
Sabouraud Dextrose Agar (SDA): Hi Media
4
Sterile swab: Hi Media
5
Autoclave: Equitron
6
Bio-safety Cabinet: Thermolab
7
Incubators: Allyone
8
Colony Counter: Lapiz
9.70%
IPA: Qualigens
3.3: Sampling
Procedure
Test
areas of 10 × 10 cm² were measured using sterile stencils. The sterile swabs
were moistened with sterile water, and samples were collected from two
different 100cm² areas of each piece of clean equipment. A total of two swab
samples from each equipment were collected using unidirectional movements—first
10 horizontal Strokes followed by 10 vertical strokes—for the determination of
total aerobic microbial count and total yeast and mould count. The swabs were
placed into separate test tubes containing 10 mL of Buffered peptone water and
transported to the microbiology laboratory.
The equipment used during the study was well
calibrated. Stencils used for measuring surface area were sterilized in an
autoclave at 121 °C and 15 psi for 15 minutes. Soybean Casein Digest Agar was
used for the isolation of bacteria, and Sabouraud dextrose agar with
chloramphenicol was used for the isolation of fungi. Growth promotion test and
Sterility checks of the swab sticks was performed as per USP<61> (United
States Pharmacopoeial Convention) 2025.
Each tube containing the swab sample was shaken for
2-3 minutes.10 ml of the sample solution was individually pipetted into 50 mL
of peptone water, mixed thoroughly, and the entire contents were filtered
through a membrane filter with a pore size of 0.45 μm. The membrane filter was
aseptically transferred onto Soybean Casein Digest Agar (SCDA) using sterile
forceps and incubated at 35 °C for 72 hours. For total yeast and mould count,
the filter was placed on Sabouraud Dextrose Agar with chloramphenicol plates
and incubated at 25 °C for 5 days.
Table 4:Cleaning Validation Study Result
|
S.No. |
Equipment |
TAMC(cfu/100cm2) (Limit:<100
cfu) |
TYMC(cfu/100cm2) (Limit:<10cfu) |
||||||
|
Sampling Location |
Result |
Sampling Location |
Result |
||||||
|
Batch1 |
Batch2 |
Batch3 |
Batch1 |
Batch2 |
Batch3 |
||||
|
1 |
Wax vessel |
From
the both side of the baffles (Mc-01) |
7 |
1 |
9 |
From the base of the vessel near discharge (Mc-02) |
0 |
1 |
0 |
|
2 |
Manufacturing Vessel |
A sample from the Teflon
flanges attached to the homogenizer .(MC-03) |
10 |
3 |
2 |
A sample from the base of
the equipment near the drain pipe (MC-04) |
1 |
0 |
1 |
|
3 |
Storage Vessel/Paste
preparation vessel |
Sample from the Base on the
other (MC-05) |
6 |
3 |
2 |
Sample from the wall of the vessel (MC-06) |
0 |
1 |
1 |
|
4 |
SS Containers |
Wall of the container (MC-07) |
20 |
23 |
14 |
Base of the container (MC-08) |
1 |
0 |
1 |
|
5 |
SS Jugs |
Base
of the jugs (MC-09) |
10 |
12 |
8 |
Base
of the jugs (MC-10) |
0 |
1 |
0 |
|
6 |
Semi- Automatic Tube Filling
Machine |
Inner surface of hopper (MC-11) |
5 |
11 |
5 |
Surface
of the stirrer (MC-12) |
1 |
0 |
1 |
|
Mean value |
8.38 |
Mean value |
0.55 |
||||||
|
Standard Deviation |
6.08 |
Standard Deviation |
0.511 |
||||||
The result of cleaning validation on Clobetasol 0.05% cream manufacturing
equipments in class D manufacturing facility in pharmaceutical Industry shows
within the acceptance limit below 100 cfu and 10 cfu respectively for Total
aerobic microbial count and Total Yeast and Mould count, after correcting the contamination variable. The
mean value of Total aerobic Microbial count and Total yeast and mould count was
found to be 8.33 and 0.55 for all three batches of used equipments
respectively. Standard deviation was found to be 6.08 and 0.511, obtained
result was not found statistically significant with respect to p- value <0.05.
Discussion
Cleaning Validation studies is an important aspect in pharmaceutical studies to identify and correct potential problems previously unsuspected, which could compromise the safety, efficacy or quality of subsequent batches of drug product produced within the equipment.
Cleaning validation study on Clobetasol 0.05% cream manufacturing equipments was performed in class D pharmaceutical Facility.The mean valueof obtained microbial count was found to be 8.33 and 0.55 for all three batches of used equipments respectively) but the obtained data was was not found statistically significant with respect to p- value <0.05.The lower microbial load on equipments might be proper cleaning procedure, controlled environment , proper fumigation programme and limited assess control in the area.
References
1.Rao N, Khan A,
Khan F,Cleaning validation in pharmaceutical industry, IJRPC 2020, 10(2),
205-214. DOI:
10.33289/IJRPC.10.2.2020.10(39).
· 2.Singh SK, Dutta K , Chaudhary R, Risk Based Study of Sample Spot Fixing for Conducting Environmental Monitoring Test in Classified Areas of Pharmaceutical Industry. Int J of Pharm Sci. 2025;3(2):146-52. [https://doi.org/10.5281/zenodo.14793799].
3.Singh SK,
Single –time point surface bioburden study at 96 hours dirty hold time study on amlodipine study amlodipine manufacturing equipment in Class D Facility (Non –Sterile). Dale
View's Journal of Clinical Pharmacology and Pharmacotherapeutics, Vol. 2 l No.
2 l December 2025
4. IPA, Indian Pharmaceutical Alliance.
Retrieved 2023-08-05
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