US Environmental Protection Agency
Office of Pesticide Programs
Office of Pesticide Programs
Mic ro bio lo gy La bo ra to ry
Environmental Science Center, Ft. Meade, MD
Standard Operating Procedure for Verification of Digital Pipettes
SOP Number: QC-19-07
Date Revised: 09-26-14

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SOP No. QC-19-07
Date Revised 09-26-14
Page 1 of 8
SOP Number
QC-19-07
Title
Verification of Digital Pipettes
Scope
Describes process for verification of digital pipettes.
Application
Pipettes are calibrated annually and are evaluated using the
gravimetric procedure as necessary.


Approval Date
SOP Developer:

Print Name:
SOP Reviewer

Print Name:
Quality Assurance Unit

Print Name:
Branch Chief

Print Name:


Date SOP issued:

Controlled copy
number:

Date SOP withdrawn:


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SOP No. QC-19-07
Date Revised 09-26-14
Page 2 of 8
TABLE OF CONTENTS
Contents	Page Number
1.
DEFINITIONS
3
2.
HEALTH AND SAFETY
3
3.
PERSONNEL QUALIFICATIONS AND TRAINING
3
4.
INSTRUMENT CALIBRATION
3
5.
SAMPLE HANDLING AND STORAGE
3
6.
QUALITY CONTROL
3
7.
INTERFERENCES
3
8. NON-CONFORMING DATA
3
9.
DATA MANAGEMENT
3
10.
CAUTIONS
3
11.
SPECIAL APPARATUS AND MATERIALS
3
12.
PROCEDURE AND ANALYSIS
4
13.
DATA ANALYSIS/CALCULATIONS
5
14.
FORMS AND DATA SHEETS
5
15.
REFERENCES
5

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SOP No. QC-19-07
Date Revised 09-26-14
Page 3 of 8
1. Definitions
Abbreviations/definitions are provided in the text.
1.	Verification failure = Verification of pipette exceeds 5% of the target
volume.
2.	Ideal volume = Target volume; actual volume being measured, corrected
for temperature using the density of water at 21.0C (0.997995 g/mL, see
section 15.1).
2. Health and
Safety
Follow procedures specified in SOP MB-01, Laboratory Biosafety. The Study
Director and/or lead analyst should consult the Safety Data Sheet for specific
hazards associated with products.
3. Personnel
Qualifications
and Training
Refer to SOP ADM-04, OPP Microbiology Laboratory Training.
4. Instrument
Calibration
Refer to SOP EQ-03 (weigh balances) for details on method and frequency of
calibration.
5. Sample
Handling and
Storage
None.
6. Quality Control
1. For quality control purposes, the required information is documented on
the appropriate form(s) (see section 14).
7. Interferences
None.
8. Non-
conforming
Data
1.	Management of non-conforming data will be specified in the study
protocol; procedures will be consistent with SOP ADM-07, Non-
Conformance Reports.
2.	Do not use pipettes if the inaccuracy exceeds 5% of the target volume.
9. Data
Management
1.	Data will be archived consistent with SOP ADM-03, Records and
Archives.
2.	Maintain an inventory of pipettes electronically using a Microsoft Excel
spreadsheet (refer to section 14). After each addition to or deletion from
the inventory, file a hard copy of the pipette inventory in the Pipette
Verification and Calibration Record Book.
10. Cautions
1.	If a pipette is dropped or damaged, it must be successfully verified using
the gravimetric procedure or recalibrated by a vendor prior to use.
2.	If a pipette fails an in-house verification assessment, it will not be used in
the laboratory and will be recalibrated by a vendor prior to use.
11. Special
Apparatus and
1. Calibrated balances. Capable of measuring 0.01 g for verifying pipettes
with volumes greater than or equal to 1 mL and 0.001 g for verifying

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SOP No. QC-19-07
Date Revised 09-26-14
Page 4 of 8
Materials
pipettes with volumes less than 1 mL.
2. Pipettes.
a.	Rainin Adjustable Volume Pipettes
b.	Gilson Microman Positive Displacement Pipettes
c.	Gilson Distriman Continuously Adjustable Volume Repetitive
Pipette
12. Procedure and
Analysis
12.1 Pipette
Verification
Requirements
a.	Annually verify and service pipettes using an ISO accredited vendor.
b.	If a pipette is dropped or broken, verify the pipette using the
gravimetric verification procedure.
c.	Record the annual verification results and if necessary, gravimetric
analysis results on the Pipette Verification Record Sheet (refer to
section 14). The Pipette Verification Record Sheet is based on the
inventory of pipettes and may change over time.
12.2 Gravimetric
Verification
Procedure
e.
f.
Record all pertinent information for the gravimetric verification
procedure on the Pipette Verification - Gravimetric Analysis Form
(refer to section 14).
In advance of testing, fill a container with de-ionized water and allow
it to equilibrate to room temperature in the same laboratory with the
balance. List the balance that will be used on the appropriate form.
Place a small Erlenmeyer flask on the balance and record its mass on
the appropriate form.
Using the pipette to be verified, aspirate an aliquot of DI water from
the sample aliquot container and dispense into the Erlenmeyer flask.
Record the mass on the appropriate form.
Follow the procedure in 12.2d-e for each subsequent sample
addition; measure at least 5 samples. Do not tare between samples.
After each new sample addition, record the mass on the appropriate
form.
Input the measurements and other appropriate information into the
Pipette Verification - Gravimetric Analysis Spreadsheet.
Verify that the percent inaccuracy is within 5%. If the percent
inaccuracy is outside of this range, remove the pipette from use until
it is repaired and calibrated by a vendor.

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SOP No. QC-19-07
Date Revised 09-26-14
Page 5 of 8

 , . (x-ideal volume)x 100
i. Percent Inaccuracy = -			
ideal volume
12.3 Pipette
Verification
Record Sheet
a.	From the data on the calibration certificate for each pipette, record
the following on the Pipette Verification Record Sheet:
i.	Verification Status: record as "pass" or "fail". Record status
as "pass" if the percent inaccuracy from gravimetric analysis
is within 5% or if vendor calibration certificate indicates
acceptable results, otherwise record as "fail".
ii.	Corrective Action: record as "yes" or "no". Record as "yes"
if the verification status is "fail" and describe the action taken
at the bottom of the page. If the verification status is "pass,"
record as "no."
b.	Generate one Pipette Verification Record Sheet annually and file in
the Pipette Verification and Calibration Log Book.
c.	The Pipette Verification Record Sheet may be completed
electronically or by hand.
13. Data Analysis/
Calculations
1. None
14. Forms and Data
Sheets
1.	Sample Pipette Verification Record Sheet
2.	Sample Pipette Inventory
3.	Test Sheets. Test sheets are stored separately from the SOP under the
following file names:
MLB Pipette Inventory QC-19-07 Fl.xlsx
Pipette Verification Record Sheet QC-19-07_F2.xlsx
Pipette Verification - Gravimetric Analysis Form QC-19-07 F3.xlsx
Pipette Verification - Gravimetric Analysis ir. 1
c , , A QC-19-07 F4.xlsx
Spreadsheet -
15. References
1. CRC Handbook of Chemistry and Physics. 93rd ed. CRC Press: Boca
Raton, FL, 2012; p 6-8.

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SOP No. QC-19-07
Date Revised 09-26-14
Page 6 of 8
Sample Pipette Verification Record Sheet
OPP Microbiology Laboratory
Verification Date(s):	Initials:
Manufacturer
Model No.
Serial No(s).
Volume
Range
In-house Verification
Volume(s)
Verification
Status1
Corrective
Action
(Y/N)


X12523D




Gilson
M10
X12649D
1-10 nL
10 nL




BH15232




Gilson
M100
GG05125
10-100 nL
100 nL


GG05127




AE10020

10 nL (1-12.5 nL)


Gilson
Distriman
BD10010
1-1250 nL
100 (iL (10-125 (iL)
900 nL (100-1250
HL)




U10048H





A0504243A






L0508039A






C0823986A






J0753884A






C0825980A




Rainin
L-1000
C0823596A
100-
1000 nL


H0101474A
1000 nL




H0100977A






J0902334A






J0908624A






D0303509A






E0301364A






A0510192A






L0509218A






C0821962A






J0750805A




Rainin
L-200
C0820542A
20-200 nL
100 nL


C0820661A




G0101809A






G0102379A






J0902612A






C0401654A





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SOP No. QC-19-07
Date Revised 09-26-14
Page 7 of 8
Manufacturer
Model No.
Serial No(s).
Volume
Range
In-house Verification
Volume(s)
Verification
Status1
Corrective
Action
(Y/N)
Rainin
L-100
C0825210A
10-100 nL
100 nL


C0825238A




A0507382A






C0822081A






J0724932A






C0822312A




Rainin
L-20
C0825315A
2-20 nL
10 nL




F0100492A






F0100448A






J0903005A






C0400398A




Rainin
L-2
H0100116A
0.1-2 nL
2 nL


H0100003A


Rainin
L-5000
D1080497A
0.5-5 mL
5 mL


Rainin
L-10000
A1058487A
1-10 mL
10 mL


Rainin
L-20000
L0931886A
2-20 mL
20 mL


Verification status = PASS if percent inaccuracy from gravimetric analysis is within 5% or if vendor calibration
certificate indicates acceptable results. If verification status = FAIL, record "yes" in Corrective Action column and
fill in Action Taken below.
Corrective Actions:
Pipette Serial Number Action Taken
1)	 	
2.)	
	

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SOP No. QC-19-07
Date Revised 09-26-14
Page 8 of 8
Sample Pipette Inventory
OPP Microbiology Laboratory
MLB Pipette Inventory
Manufacturer
Modi! No
Serial Xt/i":).


X12523D
Gils on
mm
X12649D
BK15232
Gils on
mm
GG0512"
GG05125


AE10020
Gils on
Distriman
BD10Q13
UIOCiSH


A050-243A


LQ53S93SA


CQS239SSA


J0753SS4A


C0S25980A
Raintn
L-1000
C5S23536A
HOIOU7iA
HC10G9-W
JO'90233-A
IMDS524A
DC333309A
E33C135-A
Manufacturer
Model Mo.
S^rjai Xstit


A0310I92A


L350S21SA


CCS21P52A


J37J0S35A
Ramin
L-2Q0
C0820542A
C3S23S61A
G31G1S3SA
GO 1323^3A
J09C2522A
C340165-A
Rainin
1-100
CGS252I3A
C3S2523SA


A0W3S2A


C0S223S1A


J372-1932A


C0S223I2A
Ramm
L-20
008253 ISA
F013012SA
F013C-USA
J39C3G35A
CD-3C39SA
Rainin
L-2
FECI 03116A
K01300C3A
Rainin
L-50'03
D10SD-t9~A
Rainin
L-20003
AIDSS-S'A
Raiiiin
L-23CC0
L0931SS6A

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