US Environmental Protection Agency
Office of Pesticide Programs
Office of Pesticide Programs
Microbiology Laboratory
Environmental Science Center, Ft. Meade, MD
Standard Operating Procedure for
Calibration of Kimble Class A Burets
SOP Number: EQ-06-05
Date Revised: 10-05-11

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SOP No. EQ-06-05
Date Revised 10-05-11
Page 1 of 9
EPA/OPP MICROBIOLOGY LABORATORY
ESC, Ft. Meade, MD
Standard Operating Procedure
for
Calibration of Kimble Class A Burets
SOP Number: EQ-06-05
Date Revised: 10-05-11
Initiated By:		Date: / /
Print Name:		
Technical Review:	Date: / /
Print Name:
Technical Staff
QA Review:		Date: / /
Print Name:	
QA Officer
Approved By:		Date: / /
Print Name:	
Branch Chief
Effective Date: 	/	/	
Controlled Copy No.:	
Withdrawn By:
Date: / /

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SOP No. EQ-06-05
Date Revised 10-05-11
Page 2 of 9
TABLE OF CONTENTS
Contents	Page Number
1.0 SCOPE AND APPLICATION	3
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	4
9.0 SAMPLE HANDLING AND STORAGE	4
10.0 PROCEDURE AND ANALYSIS	4
11.0 DATA ANALYSIS/CALCULATIONS	4
12.0 DATA MANAGEMENT/RECORDS MANAGEMENT	6
13.0 QUALITY CONTROL	6
14.0 NONCONFORMANCE AND CORRECTIVE ACTION	7
15.0 REFERENCES	7
16.0 FORMS AND DATA SHEETS	7

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SOP No. EQ-06-05
Date Revised 10-05-11
Page 3 of 9
1.0 SCOPE AND APPLICATION:
1.1 This protocol describes the calibration method for 10 mL and 50 mL Kimble
Class A burets. The calibration is done on an annual basis.
2.0 DEFINITIONS:
2.1	Calibration = The determination of the difference between the volume dispensed
and the expected volume.
2.2	Adjustment = The correction of the difference between the measured value and
the expected volume of the liquid displaced.
2.3	ISO = International Organization for Standardization
3.0 HEALTH AND SAFETY: Not applicable
4.0 CAUTIONS:
4.1	The water used for calibration should be weighed immediately after being
dispensed to avoid evaporation.
4.2	Burets should be inspected for chips and cracks prior to use and prior to
calibration.
5.0 INTERFERENCES:
5.1 It is important that the volumes of water are
entire calibration process can be impacted.
6.0 PERSONNEL QUALIFICATIONS:
6.1 Personnel are required to be knowledgeable
7.0 SPECIAL APPARATUS AND MATERIALS:
7.1	Kimble Class A buret (10 mL) with 0.1 mL increments.
7.2	Kimble Class A buret (50 mL) with 1.0 mL increments.
7.3	Sartorius Basic Plus Model BP21 ID (Serial Number 80904707): Weighs 0 to 40 /
80 / 210 g, reads to 0.00001 g or .01 mg, reproducibility # 0.02 / 0.05 / 0.1 mg
dispensed precisely, otherwise the
of the procedures in this SOP.

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SOP No. EQ-06-05
Date Revised 10-05-11
Page 4 of 9
(Manufacturer's Claims).
7.4	Sartorius Master1^0 Series Model LP 420 (Serial Number 81107148): Weighs 0 to
420 g, reads to 0.01 g or 10 mg, reproducibility #0.01 g (Manufacturers
Claims).
7.5	Microsoft Excel Spreadsheet (2007)
8.0 INSTRUMENT OR METHOD CALIBRATION:
8.1 The weigh balances are calibrated annually by an ISO accredited vendor and are
checked quarterly for accuracy using a reference weight set (see SOP EQ-03,
Weigh Balances).
9.0 SAMPLE HANDLING AND STORAGE: Not applicable
10.0 PROCEDURE AND ANALYSIS:
10.1	Wash and rinse the buret with de-ionized water. Place a clean 50 mL beaker on
the balance and tare it. Fill the buret with room temperature de-ionized water and
adjust the level of the water to the zero mark while allowing the rest of the buret
to become filled. Remove any air bubbles. Record the temperature of the water
on the Calibration of Kimble Class A Buret Form (see 16.1).
10.2	For the 10 mL buret, dispense into the pre-tared beaker 5 serial aliquots (2 mL
each) of water from the filled buret. Weigh the pre-tared beaker after the addition
of each 2 mL aliquot. Record results on the Calibration of Kimble Class A Buret
Form (see 16.1).
10.3	For the 50 mL buret, dispense into the pre-tared beaker 5 serial aliquots (10 mL
each) of water from the filled buret. Weigh the pre-tared beaker after the addition
of each 10 mL aliquot. Record results on the Calibration of Kimble Class A Buret
Form (see 16.1).
10.4	Weights are plotted against the independent variable of the volume reading on the
buret using Microsoft Excel.
11.0 DATA ANALYSIS/CALCULATIONS:
11.1 Simple linear regression is calculated by Microsoft Excel.
11.1.1 For the 10 mL buret, enter X values in column A of the spreadsheet and Y
values in column B, where X = theoretical value (volume in mL) and Y =

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SOP No. EQ-06-05
Date Revised 10-05-11
Page 5 of 9
actual value (weight in g).
11.1.2	In Microsoft Excel, select the "Data" tab of Excel's ribbon. Then from the
"Analysis" group, click the "Data Analysis" button. From the pop-up
menu, select "Regression" option then OK.
11.1.3	Pop-up menu:
Itagiov.ion


I Input

1 0K 1
1 Input Y Range:
1  ~~~T53



( Cancel j
I Input X Range:
i ty

I ~ Labels
PI Constant is Zero
t aelp 1
I PI Confidence Level:
: %

J Output options


I 0 Output Range:
|

1 O New Worksheet Ply:
l |

I 0 New Workbook


1 Residuals


I ~ Residuals
~ Residual Plots

1 ~ Standardised Residuals
~ Line Fit Plots

I Normal Probability


I EH formal Probability Plots


11.1.4	In the pop-up menu under Input.
	Click once in the blank cell to the right of Input Y Range, and then
highlight the column of 5 y-values in the spreadsheet.
	Click once in the blank cell to the right of Input XRange, and then
highlight the column of 5 x-values in the spreadsheet.
11.1.5	In the same pop-up menu under Output.
	Select the Output Range option.
	Click once in the blank cell to the right of Output Range, and then
highlight a single empty cell beneath the "Vol. X" column in the
spreadsheet (e.g., cell A16).
11.1.6 Select OK.

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11.1.7 Summary output
SUMMARY OUTPUT
	Regression Statistics	
Multiple R	0.999994087
R Square	0.999988173
Adjusted R Square	0.999984231
Standard Error	0.012564167
Observations	5
ANOVA

4f

SS MS
F Significance F
Regression.

1
40.04241124 40.04241124
253660.4S49 1.72618E-08
Residual

3
0.000473575 0.000157858

Total

4
40.04288481


Coeffic is nts
Standard Error t Stat
P-value
Lower 95% Upper 95% Lower 95.0%
Upper 95.0%
Intercept
X Variable 1
0.006658
1.00053
0.01317741 0.505258638
0.001986569 503.647183
0.648159495
1.72618E-08
-0.035278398 0.048594398 -0.035278398
0.99420 785 1.0 0685215 0.9942 0785
0.048594398
1.00685215
11.1.8 Repeat steps 11.1.1 through 11.1.7 for the 50 mL buret.
11.2 The correction factor, or adjustment, for any volume (y) dispensed by the buret is
obtained by inserting the volume reading (in mL) on the buret into the following
equation:
y = mx + b; where y = corrected volume, m = slope (x variable 1 coefficient), x =
volume reading on buret, and b = y-intercept (intercept coefficient). The numbers
are rounded to the nearest tenth of an mL.
11.2.1	A table within the spreadsheet generates a correction factor for each
volume evaluated using the aforementioned calculation. The average
correction factor is calculated and used to determine the correction factor
for the buret.
11.2.2	Place a label displaying the date of calibration and the correction factor
(even if it is zero) for any volume dispensed by the buret around the top of
the buret.
12.0 DATA MANAGEMENT/RECORDS MANAGEMENT:
12.1 Data will be recorded promptly, legibly and in indelible ink on the Calibration of
Kimble Class A Buret Form. The accompanying spreadsheet will be archived
with the raw data sheet. Completed forms are archived in notebooks kept in
secured file cabinets in the file room D 217. Only authorized personnel have
access to the secured files. Archived data are subject to OPP's official retention
schedule contained in SOP ADM-03, Records and Archives.
13.0 QUALITY CONTROL:
SOP No. EQ-06-05
Date Revised 10-05-11
Page 6 of 9
13.1 The calibration of burets is performed annually and the information is

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SOP No. EQ-06-05
Date Revised 10-05-11
Page 7 of 9
documented on the appropriate record form(s) (see 16.1).
14.0 NONCONFORMANCE AND CORRECTIVE ACTION:
14.1	Burets exhibiting chips and cracks will not be used in the laboratory and will be
discarded.
14.2	When routinely using the burets in the laboratory to dispense liquids, analysts
must record the volume dispensed by the buret plus the corrected value, or
adjustment, for that volume.
15.0 REFERENCES: None
16.0 FORMS AND DATA SHEETS:
16.1	Calibration of Kimble Class A Buret Form
16.2	Example of Regression Analysis Spreadsheet

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SOP No. EQ-06-05
Date Revised 10-05-11
Page 8 of 9
Calibration of Kimble Class A Buret
OPP Microbiology Laboratory
Date/Initials Temperature of Water
10 mL Buret ID
Volume X (mL Buret)
Weight Y (gm)
2 ml

4 mL

6 mL

8 mL

10 mL

50 mL Buret ID
Volume X (mL Buret)
Weight Y (gm)
10 mL

20 mL

30 mL

40 mL

50 mL

The corrected value for any volume (y) dispensed by the buret is obtained by inserting the volume reading (in mL)
on the buret into the following equation: y = mx + b; where y = corrected volume, m = slope (x variable 1
coefficient), x = volume reading on buret, and b = y-intercept (intercept coefficient). The numbers are rounded to
the nearest tenth of an mL.

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SOP No. EQ-06-05
Date Revised 10-05-11
Page 9 of 9
Example of Regression Analysis Spreadsheet
Calibration Dale;
::::::
Almost Msm:
J. Smith
Temperature of water:
23=C
Tkermsmeisr *:
#12345
Sum Volume:
iOmL
Burst ID:
#1
	

4

6

i
j v.
10

C;r
V;l. X = 2 ml
Vol. X - - rrX
Vd X - 6 niL
Yel.X-SmL
Yd. X = 1C raL
Correction Facmr
o.o
o.o
li-g. Correction Factor
0.0
Tb deteraniie collected y value: j = an -
S^.aiARY 01
P.
R. sqiiar?
Adjust id E. Squar
Standard Error
Observations
AXOYA
_	SS	MS	f
E-ssr^sicn

- -CC~24i;2- 2:5&5
; 5 : ":5:se-:?
Residual
3 ... - :
	: :

Total
4


5
C:-"': r:
jifittupr
X*"3iat^ 1

: :C:25S'ciS : 
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