Standard Operating Procedure for Media and Reagent Preparation: Assigning Prep and Sterilization Run Numbers SOP Number: QC-15-04 Date Revised: 11-06-08


mm s r4
US Environmental Protection Agency
Office of Pesticide Programs
Office of Pesticide Programs
Microbiology Laboratory
Environmental Science Center, Ft. Meade, MD
Standard Operating Procedure for
Media and Reagent Preparation: Assigning Prep
and Sterilization Run Numbers
SOP Number: QC-15-04
Date Revised: 11-06-08

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SOP No. QC-15-04
Date Revised 11-06-08
Page 1 of 8
EPA/OPP MICROBIOLOGY LABORATORY
ESC, Ft. Meade, MD
Standard Operating Procedure
For
Media and Reagent Preparation: Assigning Prep and Sterilization Run Numbers
SOP Number: QC-15-04
Date Revised: 11-06-08
Initiated By:
Print Name:
Technical Review:
Print Name:
Technical Staff
QA Review:
Print Name:
QA Officer
Approved By:
Print Name:
Branch Chief
Date: / /
Date: / /
Date: / /
Date: / /
Effective Date:
/ /
Controlled Copy No.:
Withdrawn By:
TABLE OF CONTENTS
Date: / /

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SOP No. QC-15-04
Date Revised 11-06-08
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Contents	Page Number
I.0	SCOPE AM) APPLICATION	2
2.0	DEFINITIONS	3
3.0	HEALTH AND SAFETY	3
4.0	CAUTIONS	3
5.0	INTERFERENCES	3
6.0	PERSONNEL QUALIFICATIONS	3
7.0	SPECIAL APPARATUS AND MATERIALS	 3
8.0	INSTRUMENT OR METHOD CALIBRATION	3
9.0	SAMPLE HANDLING AND STORAGE	3
10.0	PROCEDURE AM) ANALYSIS	4
II.0	DATA ANALYSIS/CALCULATIONS	5
12.0	DATA MANAGEMENT/RECORDS MANAGEMENT	5
13.0	QUALITY CONTROL	5
14.0	NONCONFORMANCE AND CORRECTIVE ACTION	5
15.0	REFERENCES	5
16.0	FORMS AND DATA SHEETS	5
1.0 SCOPE AND APPLICATION:

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SOP No. QC-15-04
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1.1 This standard operating procedure describes how to assign media/reagent
preparation and sterilization batch control numbers. Tracking media and reagents
from preparation to use must be thoroughly documented.
2.0 DEFINITIONS: None
3.0 HEALTH AND SAFETY: Not applicable
4.0 CAUTIONS: None
5.0 INTERFERENCES:
5.1 The suffix on all preparations is critical for the tracking of all preparations and its
assignment must be made by following the SOP explicitly.
6.0 PERSONNEL QUALIFICATIONS:
6.1 Personnel must be knowledgeable of the procedures for assigning preparation and
sterilization batch control numbers. Documentation of training and
familiarization with this SOP can be found in the training file for each employee.
7.0 SPECIAL APPARATUS AND MATERIALS:
7.1 Autoclave #1 located in room B206, Amsco Eagle 3000 Scientific Series, Model
E3031-S-1, Serial No. 0105898-25
7.2 Autoclave #2 located in room B204, Amsco Eagle 3000 Scientific Series, Model
E3031-S-1, Serial No. 0108298-11
7.3 Autoclave #3 located in room B207, Amsco Century Series Scientific Gravity
Sterilizer (16x16x26"), Model SG-116, Serial No. 0121198-15
7.4 Autoclave #4 located in room B202, Amsco Century Series Scientific Gravity
Sterilizer (20x20x26"), Model SG-120, Serial No. 0103000-16
7.5 Autoclave # 5 located in room D122, Tuttnauer Prevacuum Steam Heated
Autoclave with Vertical Sliding Door and Steam Generator (52x72x51"), Model
5596-EP-1V, Serial No. 2311036
8.0 INSTRUMENT OR METHOD CALIBRATION: Not applicable
9.0
SAMPLE HANDLING AND STORAGE:

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SOP No. QC-15-04
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9.1 Media and reagents are subject to proper storage as specified by the manufacturer.
10.0 PROCEDURE AND ANALYSIS:
10.1	When preparing media and reagents, completion of the Media/Reagent
Preparation Sheet and Media/Reagent Preparation Log Form is required (see
16.0).
10.2	The Media/Reagent Preparation sheet identifies information relevant to the
preparation of the media or reagent. The preparation sheet may also be used for
non-media items that require a preparation number such as glass slide carriers,
porcelain carriers, and steel carriers.
10.3	All boxes on the Media/Reagent Preparation sheet must be filled out with the
appropriate information for that section. If a section is not applicable to the item
being prepared, place N/A (Not Applicable) in that section.
10.4	Record the storage requirement for the media at the bottom of the page. If the
material is to be used immediately and requires no storage, then place N/A in that
section.
10.5	All media and reagents will be assigned a preparation control number.
10.5.1	The preparation control number consists of two parts: 1) the first seven
digits represent the date the medium or reagent was prepared:
P-MMDDYY where P=Prepared, MM=month, DD=day, and YY=the
last two digits of the calendar year; and 2) the suffix, where the digits
after the dash act as a counter for the number of preparations made on
the same date. For example, the first preparation made on November
6, 2008 would have the control number PI 10608-01. The next item
prepared on that same day would have a suffix of -02; the third
preparation made on that same day would have a suffix of -03, etc.
10.5.2	For all media and reagents that are sterilized by autoclaving, a
sterilization batch number is assigned. The procedure for assigning
the sterilization batch number follows similar notation for assigning
preparation control numbers. The sterilization batch number consists
of two parts: 1) the first seven digits represent the date the batch was
sterilized: S-MMDDYY where S=Sterilization, MM=month, DD=day,
and YY=the last two digits of the calendar year; and 2) the suffix
where the first digit after the dash indicates the autoclave used and the

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SOP No. QC-15-04
Date Revised 11-06-08
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next two digits act as a counter for the number of preparations made
on the same date. For example, the first batch sterilized on November
6, 2008 in autoclave 1 (Room B206) would have the control number
S-l 10608-101. The next batch sterilized on that same day and same
autoclave would have a suffix of -102; the third batch sterilized would
have a suffix of -103; etc.
11.0 DATA ANALYSIS/CALCULATIONS: None
12.0 DATA MANAGEMENT/RECORDS MANAGEMENT:
12.1 Data will be recorded promptly, legibly, and in indelible ink on the appropriate
forms. Completed forms are archived in notebooks kept in secured file cabinets
in the file room D217. Only authorized personnel have access to the secured
files. Archived data is subject to OPP's official retention schedule contained in
SOP ADM-03, Records and Archives.
13.0 QUALITY CONTROL:
13.1 For quality control purposes, the required information is documented on the
appropriate form(s) (see 16.0).
14.0 NONCONFORMANCE AND CORRECTIVE ACTION:
14.1 If a preparation control number or sterilization batch number is missing and
cannot be determined from the Media/Reagent Preparation Sheet (see 16.1), or the
Daily Sterilization Record Information Log Form (see 16.2), the media or reagent
will not be used in any testing procedure and will be discarded.
15.0 REFERENCES: None
16.0 FORMS AND DATA SHEETS:
16.1	Media/Reagent Preparation Sheet
16.2	Media/Reagent Preparation Log Form
16.3 Daily Sterilization Record Information Log Form

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16.1
SOP No. QC-15-04
Date Revised 11-06-08
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MEDIA/REAGENT PREPARATION SHEET
OPP MICROBIOLOGY LABORATORY
Media/Reagent
Name:
Amount Prepared:
Prep #:
Preparation Date/Initials:
pH Meter #:
Dispenser: (circle one)
Water Bath #(s):
BSC #(s):
Sterilization #:

Hamilton Oxford Hand NA



Media/Chemical Ingredients:
Control No:
Amount Required:
Amount Measured:












Preparation/Modifications/Notes:
Required pH:
pH Before Autoclaving with Medium
Cooled to Room Temperature:
Final pH After Autoclaving with
Medium Cooled to Room Temperature:

pH:
Temperature:
pH:
Temperature:
Volume of Acid/Base added lo obtain final pH:
Sterility/Viability Test Results
Sterility
Viability

Pass

Fail
Pass

Fail
Storage of Reagent/Media:
Media Requested bv:
Media Prepared for: ATP R&D PIP

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SOP No. QC-15-04
Date Revised 11-06-08
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16.2
Media/Reagent Preparation Log Form
OPP Microbiology Laboratory
Media Information
Date/Initials
Preparation Number
Type of Media/Reagent
Amount

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SOP No. QC-15-04
Date Revised 11-06-08
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16.3
Daily Sterilization Record Information Log Form
OPP Microbiology Laboratory

Cycle1
Unit Temp.2
Max. Thermometer3
Chemical Indicator Strips
Items Processed
Sterilization No.5
Init.
Type
Time
Min.
Max.
Temp.
Serial #
Lot No.
Exp. Date
Result4








































































































































































'Record the cycle as "G" = gravity, "L" = liquid under Type and the duration of the cycle in minutes under Time.
2Record the maximum and minimum temperature achieved during the sterilize phase of the cycle as indicated by the autoclave printout (Unit).
3Record the corrected value for the maximum registering thermometer (Max.) and the serial number of the thermometer.
4Record the results of the chemical indicator strips as "P" for pass or "F" for fail.
5The Sterilization No. indicates the date as well as the unit location and the run number.

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