Protocol cum report for efficacy qualification of uv light

LOCATION: DRY POWDER INJECTION BLOCK

AND MICROBIOLOGY LAB.

Protocol No.
Date of Qualification
Supersedes
Protocol Contains Pages

PROTOCOL CONTENTS

Point No.	Section Title	Page No.
1.0	Protocol Approval
2.0	Objective
3.0	Scope
4.0	Responsibility
5.0	Reason For Validation
6.0	Site of Study
7.0	Type of Validation
8.0	Acceptance Criteria
9.0	Methodology For Qualification
10.0	List of Equipments To Be Used For Qualification
11.0	Qualification Procedure
12.0	Revalidation Criteria
13.0	Frequency of Validation
14.0	Non Compliance
15.0	Exhibits
16.0	References
17.0	Documents To Be Attached
18.0	Any Changes Made Against The Formally Agreed Parameters
19.0	Review (Inclusive of Follow Up Action, If Any)
20.0	Recommendation
21.0	Authorization
22.0	Abbreviations

1.0 PROTOCOL APPROVAL

INITIATED BY:

DESIGNATION	NAME	SIGNATURE	DATE
QA EXECUTIVE

REVIEWED BY:

DESIGNATION	NAME	SIGNATURE	DATE
MANAGER PRODUCTION
MANAGER ENGINEERING
MANAGER QC
MANAGER QA

APPROVED BY:

DESIGNATION	NAME	SIGNATURE	DATE
GM WORKS

AUTHORIZED BY:

DESIGNATION	NAME	SIGNATURE	DATE
HEAD QA

2.0 OBJECTIVE:
To provide documented evidence, that the UV light radiation used is suitable to control microorganisms on the surface of the material (if present) which passes through the pass box.
To provide documented evidence, that the UV light radiation used is suitable to control microorganisms on the surface of the sterilized garments and inside of the garment cubicle (if present).
To provide method for qualification of the Bactericidal and Fungicidal Efficacy of UV light radiation used in the pass boxes and garment cubicles.

3.0 SCOPE:
The protocol covers all aspects of Qualification of UV Light radiation (used in the pass boxes and garment cubicles) in the DPI and Microbiology lab
The protocol is applicable for checking the Bactericidal and Fungicidal Efficacy of UV light radiation.

4.0 RESPONSIBILITY:
The validation group, comprising of a representative from each of the following departments, shall be responsible for the overall compliance of this protocol cum report:

DEPARTMENTS RESPONSIBILITIES
Quality Assurance Preparation, review, approval and compilation of the Qualification Protocol cum Report.
Co-ordination with Microbiology, Production and Engineering to carryout Qualification Study.
Monitoring of Qualification activity.
Production Review of Qualification Protocol cum Report.
To co-ordinate and support Qualification activity.
Engineering Review of Qualification Protocol cum Report.
To co-ordinate and support Qualification activity.
Quality Control Review of Qualification Protocol cum Report.
Sampling & Microbiological Testing / Analysis.

5.0 REASON FOR VALIDATION:
After completion of the Qualification of the pass boxes and garment cubicles in the area, it is imperative to perform the Qualification of UV Light Radiations. The study will establish that the Bactericidal and Fungicidal Efficacy of UV light radiation used in the pass boxes and garment cubicles is effective to control the micro – organisms (if any), and is working, as desired.

6.0 SITE OF STUDY:
Manufacturing blocks i.e. DPI and Microbiology lab
7.0 TYPE OF VALIDATION:
Concurrent Validation.

8.0 ACCEPTANCE CRITERIA:
The UV Light Radiation must be able to kill the challenged microorganisms, and the contact time should be not more than 15 minutes.

9.0 METHODOLOGY FOR QUALIFICATION:

9.1 SOP / SPECIFICATIONS TO BE FOLLOWED:
Procedure for Preparation of Culture Dilutions as per SOP No.QC/211-00.
Procedure for Preparation of Media as per SOP No.QC/208-00.
Sterilization of Glass Wares and Apparatus as per SOP No.QC/234-00.
9.2 MATERIALS REQUIRED:
Slants of Nutrient agar and Sabouraud’s Chloramphenicol Agar.
Sterile 0.9% w/v saline.
Cultures of Bacillus subtilis, E. coli, two common isolates of the area and C. albicans.
Sabouraud Chloramphenicol Agar (SCA) and Soyabean Casein Digest Agar(SCDA).
Sterile Petri-dishes, pipettes, etc.
9.3 BRIEF PROCEDURE:
Sterilize the glass wares. Prepare the media & sterilize the media. Transfer from stock, the specified microbial cultures on the Nutrient Agar Plates and Incubate.
Bring the pre incubated media plates at the test location. Remove the plate lid and expose the SCDA / SCA plates as per procedure inside the pass box and garment cubicle. Bring the exposed plates to Microbiology area and incubate for specified time. Observe the plates after 05 days. The exposed plates should pass as per pre defined acceptance criteria.

9.4 INTENSITY OF UV LIGHT USED:
251 nm (15 Watts / Hour)

10.0 LIST OF EQUIPMENTS TO BE USED FOR QUALIFICATION:
S.No. EQUIPMENT NAME EQUIPMENT ID No. LOCATION No. of UV Tubes
1. Garment Storage Cabinet Microbiology Lab 01
2. Static Pass Box Microbiology Lab 01
3. Dynamic Pass Box Microbiology Lab 01
4. Garment Storage Cabinet DPI Block 01
5. Static Pass Box DPI Block 01
6. Dynamic Pass Box DPI Block 01
7. Dynamic Pass Box DPI Block 01

11.0 QUALIFICATION PROCEDURE:
11.1 PROCEDURE FOR CHECKING THE EFFICACY OF UV LIGHT USED IN THE GARMENT CUBICLE:
Sterilize the glass wares, pipettes, as per SOP No.QC/234-00.
Prepare the media as per the SOP No.QC & sterilize the media as per SOP No. QC.
Check the total burning hours of UV light before starting the validation studies. If the total burning hour is more than 7000 hours, replace the UV lamp.
Before starting validation studies check the intensity of UV light.
Transfer from stock, the following microbial cultures on to the Nutrient Agar and Incubate as given in the table-1.
For C. albicans use Sabouraud Chloramphenicol Agar (SCA).

Table –1

Name of organism	Media used	Temperature for incubation	Duration of incubation
E.coli	SCDA	30⁰-35⁰ C	24-48 hr
B.subtilis	SCDA	30⁰-35⁰ C	24-48 hr
C.albicans	SCA	20⁰-25⁰ C	48-72 hr
Common isolate –I	SCDA	30⁰-35⁰ C	24-48 hr
Common isolate-II	SCDA	30⁰-35⁰ C	24-48 hr

Bring the pre incubated media plates at the test location. Remove the lid of the SCDA plates and put one plate each inside Garment Cubicle as per below mentioned locations:

P1 P2

P3 P4

P1= Left corner back, P2= Right corner back, P3 = Left corner front,
P4 = Right corner front, P5 = center of the LAF

Switch on the UV light and note down the time. After 05 minutes, switch-off the UV light and remove the plate.
Mop the platform / surface with 70% IPA and switch on the UV light for about 30 minutes.
Similarly repeat the procedure with SCA plates.
Repeat the above experiment for the other sets of the plates using SCDA and SCA separately marked as “10 min”, “15 min”, “30 min” and “45 min”.
Bring the sampled media plates to the microbiology lab.
Incubate the plates at 30-350C for 5 days.
Incubate the plates at 20-250C for 5 days.
FOR POSITIVE CONTROL:
For positive controls prepare culture dilutions as per SOP No.QC/211-00 having 10 – 100 cfu/ml.
Inoculate the SCDA plate with culture of B. subtils .and incubate at 30-35°C for 5
days.
Inoculate the SCA plate with culture of C.albicans .and incubate at 20-25°C for 5
Days.

FOR NEGATIVE CONTROL:
For negative control incubate one un-inoculated plate of SCA and SCDA at 20-25°C for 72 hrs.
After 72 hrs transfer the plates to 30-35°C and incubate for 48 hrs.
Observe the plate after completion of incubation and record the results in Exhibit-I.

OBSERVATION AND INTERPRETATION OF RESULTS:
Observe the plates after 72 hrs. and record if any fungal colony is found.
Observe the plates at the end of incubation and count the total no of cfu/plate and
record it in the Exhibit – I.
The plate of negative control should remain clear after completion of incubation.
The plate of positive controls should show growth within 48-72 hrs after incubation.

11.2 PROCEDURE FOR CHECKING THE EFFICACY OF UV LIGHT USED IN THE PASS BOX:
Sterilize the glass wares, pipettes, as per SOP No.QC
Prepare the media as per the SOP No.QC & sterilize the media as per SOP No. QC/234-00.
Check the total burning hours of UV light before starting the validation studies. If the total burning hour is more than 7000 hours, replace the UV lamp.
Before starting validation studies check the intensity of UV light.
Transfer from stock, the following microbial cultures on to the Nutrient Agar and Incubate as given in the table-1. For C. albicans use Sabouraud Chloramphenicol Agar (SCA).

Table –1

Name of organism	Media used	Temperature for incubation	Duration of incubation
E.coli	SCDA	30⁰-35⁰ C	24-48 hr
B.subtilis	SCDA	30⁰-35⁰ C	24-48 hr
C.albicans	SCA	20⁰-25⁰ C	48-72 hr
Common isolate –I	SCDA	30⁰-35⁰ C	24-48 hr
Common isolate-II	SCDA	30⁰-35⁰ C	24-48 hr

Take 5 plates of Soya bean Casein Digest Agar and mark them as “05 min”, “10 min”, “15 min”, “30 min” and “45 min”.
Add 0.1 ml culture suspension having 100 cfu/ml to each of the plate.
Spread the culture with the help of glass rod (‘L’ shaped).
Keep one un-inoculated plate as negative control and one inoculated plate as positive control. Mark the positive control plate as “positive control” un exposed, Date of test and the Name of organism. Mark negative control plates as “Negative control”, Date of test.
Mop the outer and inner surfaces of the pass box with 70% IPA and switch on the UV light for 30 minutes before starting the validation.
Remove the lid of the SCDA plates and put one plate each in the pass box as per below mentioned locations:

P1 P2

P3 P4

P1= Left corner back, P2= Right corner back, P3 = Left corner front,
P4 = Right corner front, P5 = center of the LAF

Switch on the UV light and note down the time. After 05 minutes, switch-off the UV light and remove the plate.

Mop the platform with 70% IPA and switch on the UV light for about 30 minutes.
Similarly repeat the procedure with SCA plates.
Repeat the above experiment for the other sets of the plates using SCDA and SCA separately marked as “10 min”, “15 min”,“30 min” and “45 min”.
Bring the sampled media plates to the microbiology lab.
Incubate the plates at 30-350C for 5 days.
Incubate the plates at 20-250C for 5 days.

FOR POSITIVE CONTROL:
For positive controls prepare culture dilutions as per SOP No.QC/211-00 having 10 – 100 cfu/ml.
Inoculate the SCDA plate with culture of B. subtils .and incubate at 30-35°C for 5
days.
Inoculate the SCA plate with culture of C.albicans .and incubate at 20-25°C for 5
days.

OBSERVATION AND INTERPRETATION OF RESULTS:
Observe the plates after 72 hrs. and record if any fungal colony is found.
Observe the plates at the end of incubation and count the total no of cfu/plate and
record it in the Exhibit – II.
The plate of negative control should remain clear after completion of incubation.
The plate of positive controls should show growth within 48-72 hrs after incubation.

12.0 REVALIDATION CRITERIA:
If there is change in the type / size of UV light tube used.
If there is change in the intensity of the UV light.

13.0 FREQUENCY OF VALIDATION:
Once

14.0 NON COMPLIANCE:
In case of any deviation observed during Hold Time Study, inform to Department Head and Head Quality for necessary action.
Document the deviation detail in observed deviation section.
The Head Quality and the Department Head will study the impact of deviation. If deviation is acceptable and it does not have an impact on operation as well as on overall performance of the study, prepare final conclusion.

15.0 EXHIBITS:
EXHIBIT – I

UV LIGHT EFFICACY REPORT FOR GARMENT CUBICLE:

Area		Report No
Time		Date of Sampling
Media Used: Soya bean Casein Digest Agar		Media Preparation Date
Media pH		Test conducted by
Media lot No	Date	+ve Control	-ve Control

Grade –A class 100(Under LAF)
Limit-For bacteria<1 cfu/Plate. Fungi: Nil

S. No.

SAMPLE LOCATION

CFU / PLATE

BACTERIA

FUNGI

05 min.

10 min.

15 min.

30 min.

45 min.

05 min.

10 min.

15 min.

30 min.

45 min.

1

2

3

4

5

6

Observation: Bio-burden is found within / out of specified limit.

Compiled By:
(QA)
Sign & Date

Inference:
__________________________________________________________________________________________________________________________________________________________________________________________________________________________________________

Reviewed By: _______________

(Manager QA)

Sign / Date:

EXHIBIT – II

Area		Report No
Time		Date of Sampling
Media Used: Soya bean Casein Digest Agar		Media Preparation Date
Media pH		Test conducted by
Media lot No	Date	+ve Control	-ve Control

Grade –A class 100(Under LAF)
Limit-For bacteria<1 cfu/Plate. Fungi: Nil

S. No.

SAMPLE LOCATION

CFU / PLATE

BACTERIA

FUNGI

05 min.

10 min.

15 min.

30 min.

45 min.

05 min.

10 min.

15 min.

30 min.

45 min.

1

2

3

4

5

6

Observation: Bio-burden is found within / out of specified limit.

Compiled By:
(QA)
Sign & Date

Reviewed By: _______________

(Manager QA)

Sign / Date:

16.0 REFERENCES:
The Principle Reference is the following:
Schedule – M – “Good Manufacturing Practices and Requirements of Premises, Plant and Equipment for Pharmaceutical Products.”
WHO Essential Drugs and Medicines Policy, QA of Pharmaceuticals, Vol 2 – Good Manufacturing Practices and Inspection.

The following references are used to give addition guidance:
FDA/ISPE Baseline Pharmaceutical Engineering Guide-Volume 5:- Commissioning and Qualification Guide, First Edition / March 2001.
Code of Federal Regulations (CFR), Title 21, Part 210, Current Good Manufacturing Practice (cGMP) in Manufacturing, Processing, Packing, or Holding of Drugs, General. April 1, 1998.
Code of Federal Regulations (CFR), Title 21, Part 211, Current Good Manufacturing Practice (cGMP) for Finished Pharmaceuticals, April 1, 1998.
EU Guide to Good Manufacturing Practice, Part 4, 1997.
GAMP Guide, Validation of Automated Systems in Pharmaceutical Manufacture, Version 4.0, December 2001.

17.0 DOCUMENTS TO BE ATTACHED:
Sterility Test Reports of Incubated Media Plates.

18.0 ANY CHANGES MADE AGAINST THE FORMALLY AGREED PARAMETERS:

19.0 REVIEW (INCLUSIVE OF FOLLOW UP ACTION, IF ANY):

20.0 RECOMMENDATION:

21.0 AUTHORIZATION:

Manager Production Manager Engineering GM Works Head QA
(Sign / Date) (Sign / Date) (Sign / Date) (Sign / Date)

22.0 ABBREVIATIONS:
DPI : Dry Powder Injection

QC : Quality Control

Pvt. : Private

Ltd. : Limited

QA : Quality Assurance

cfu : Colony Forming Unit