CALIBRATION OF Piston Pipette and Buritte PDF

Identification:
Procedure for calibration

PROC-TC-012
Central Organization for Standards and Revision: 1
Quality Control
(COSQC)- Metrology Department Valid after:
WORK PROCEDURE
FOR
CALIBRATION OF Piston Pipette and Buritte
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Table of CONTENTS Page No.
1 Purpose 3
2 Scope 3
3 Normative References 3
4 Terms And Definition 3
5 Principle of operation 5
6 Design 5
7 Marking 8
8 Apparatus 9
9 Test conditions 10
10 Gravimetric Procedure for calibration 11
11 Calculations 16
12 Calculations for Measurement Uncertainty 16
13 Associated Documents 16
APPENDIX A 19
APPENDIX B 30
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1.0 Purpose
Purpose of this Lab Calibration Procedure is to provide guideline for the calibration of
volumetric piston pipette.
2.0 Scope
 Calibration of piston pipettes (as ISO 8655-6) from 1µl to 10ml:

 with fixed volume
 with adjustable volume
 Calibration of piston burette (as ISO 8655-3) up to 100ml
3.0 Normative references
 ISO 8655-1,2002 Terminology, general requirements and user recommendations

 ISO 8655-2,2002 Piston pipettes
 ISO 8655-3,2002 Piston Burette
 ISO 8655-6,2002 Gravimetric methods for the determination
 Operating Manual of Mettler balance (microbalance xpe26pc, xp505c).
 Operating Manual of Sartorius balance (Te214S).
 ISO/TR 20461:2000: Determination of uncertainty for volume measurements made using
the gravimetric method
 Quality Manual of Calibration Services ISO/IEC 17025.

4.0 Terms and definitions
For the purpose of this Lab Calibration Procedure following definitions shall apply:
4.1 Maximum permissible error

Upper or lower permitted extreme value for the deviation of the dispensed volume from the
nominal volume or selected volume of a piston-operated volumetric apparatus
4.2 Systematic error
The difference between the dispensed volume and the nominal volume or selected volume of
the piston-operated volumetric apparatus
4.3 Random error
The scatter of the dispensed volumes around the mean of the dispensed volumes
4.4 Uncertainty of measurement
The parameter, associated with the dispensed volume, that characterizes the dispersion of the
Volumes that could reasonably be attributed to the dispensed volume
4.5 Nominal volume
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The volume specified by the manufacturer and used for identification and for indication of the
measuring range
4.6 Selected volume
The volume set by the user, in order to dispense a volume chosen from the useful volume
range of a variable-volume piston-operated volumetric apparatus
4.7 Useful volume range
That part of the nominal volume which allows dispensing under observance of the maximum
permissible errors as specified in the applicable part of ISO 8655
4.8 Dead air volume
(piston-operated pipettes with air interface) air volume between the lower part of the piston
and the surface of the liquid
4.9 Dead liquid volume
(positive displacement pipettes, burettes, dilutors and dispensers) amount of liquid which does
not belong to the dispensed volume and which is contained during operation in aspiration or
expelling tubes, valves and within the cylinder
4.10 Adjustment
(piston-operated volumetric apparatus) manufacture of an apparatus within appropriate
tolerances, or the supplier's setting of the apparatus, ensuring the metrological performance,
as specified in the applicable part of ISO 8655, for a reference temperature of 20oC.
4.11 User adjustment
( piston-operated pipette) adjustment employing only the means at the disposal of the user
4.12 Calibration
(piston-operated volumetric apparatus) set of operations that establish the relationship
between the dispensed volume and the corresponding nominal or selected volume of the
apparatus 20oC.
4.13 Calibration certificate
Document issued after the calibration of a piston-operated volumetric apparatus, reporting the
result of calibration
4.14 Traceability
The property of a measurement result whereby it can be related to appropriate standards
generally national or international standards through an unbroken chain of comparisons.
4.15 Measurement Standards
A Material measure, measuring instrument or system intended to define, realize, conserve or
reproduce a unit by one or more known value of a quantity, in order to transmit them to other
measuring instruments by comparison.
4.16 UUC (Unit under Cal)
Unit under Cal is the instrument, which is being calibrated by this method statement.
4.17 Reference Standard
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A standard generally of a highest metrological quality available from which measurements

made at a given location are derived.
4.18 Internal Uncertainty or Type “A”
The component of uncertainty, which is caused by type “A” errors and is evaluated from the
measurement results using statistical methods.
4.19 External Uncertainty or Type “B”
It is the component of uncertainty, which is caused by Type “B” errors and is evaluated by
non-statistical methods from the external factors having influence on the measurement
4.20 Repeatability
The degree of agreement between results of successive measurements of the same measurand
carried out under identical conditions, with identical methods and over a short period of time.
4.21 Resolution
Smallest difference that can be meaningfully distinguished (e.g. a change of one (1) in the last
place of a digital display)
Extreme values of error permitted by specification / class
4.23 Dispense
To deliver the sample.
4.24 Ambient pressure
The pressure surrounding a device, often equal to the prevailing atmospheric pressure.
5 Principle of operation
5.1 for Piston Pipette
The tip made of plastic or glass is attached to the piston pipette. With the piston at the lower
aspiration limit, the tip is dipped into the liquid to be dispensed as a measured volume. When
moved to the upper aspiration limit, the piston aspirates the liquid. The liquid volume to be
dispensed is then expelled by depressing or sliding the piston between the volume-defining limits.
Some air-displacement piston pipettes have an extra air volume which can be used to expel the last
drop of liquid. See also Figure 1.
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Type D
Type A
Positive (direct) displacement Air displacement
a) Single-channel piston pipettes b) Multi-channel piston pipette
Figure 1 — Piston pipettes
5.2 For Piston Burette

piston burettes are required to dispense volumes of liquids until external criteria such as pH or
conductivity are met, at which point it is necessary to know the accurate volume dispensed.
The piston can be operated manually, electrically, pneumatically or hydraulically. The volume
delivered can be indicated mechanically or by electronic means. The drive, the piston and the
cylinder can be one unit, or modular to permit the use of different pistons and cylinders Prior to
delivery, the piston system is charged by aspiration of liquid from a reservoir. After air-bubble-free
filling of the system, movement of the piston in one direction dispenses the liquid whose volume is
to be measured; movement in the other direction recharges the system from the reservoir (see
Figure 2).
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Figure 2 — Schematic drawing of a piston burette

Piston burettes may or may not be equipped with valves or may have several piston/cylinder systems which
dispense continuously
6 Design
6.1 Types of piston pipette

A piston pipette may be designed as follows:
1. fixed volume, designed by the manufacturer to dispense only its nominal volume, for
example 100 l.
2. Variable volume, designed by the manufacturer to dispense volumes selectable by the user
within its specified useful volume range, for example between 10 l and 100 l and the
piston may either have a body of air contained between the piston and the surface of the
liquid (air displacement - type A) or be in direct contact with the surface of the liquid
(positive or direct displacement – type D).
In the case of the type D piston pipette, either the plunger or the capillary or both may be reusable
(type D1) or disposable (type D2). See Figure 1 for details.
6.2 Adjustment
1. For Piston Pipette
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 A piston pipette shall be adjusted by its manufacturer for the delivery (Ex) of its nominal
volume (or selected volume, in the case of a variable-volume model), for the standard
reference temperature of 20 oC, a relative air humidity of 50 % and a barometric pressure of
101 kPa(1.01 mbar) using grade 3 water as specified in ISO 3696.
 Some piston pipettes have provision for adjustment by the user when, for example, it is found
in routine calibration that the volume delivered is not within specification. Such user
adjustment shall be made according to the manufacturer's instructions and by reference to
the gravimetric test method specified in ISO 8655-6. Any piston pipette so adjusted shall
have clear, visible evidence that the initial adjustment has been modified. This information
shall also be recorded on any certificate of conformity and the procedure should be included
in the user's quality manual.
 Some piston pipettes are designed to have their factory-preset adjustment altered by the user
so that they will dispense their specified volume when used with liquids with physical
properties differing from those of water. Such user adjustment shall be made according to
the manufacturer's instructions and by reference to the gravimetric test method specified in
ISO 8655-6, but using the selected liquid instead of water. If the user readjusts the piston
pipette, it shall be clearly and unequivocally indicated on the outside of the piston
pipette that readjustment has been effected. The user shall mark the outside of the piston
pipette with the name of the liquid for which the nominal volume now applies.
2. For Piston Burette

A piston burette shall be adjusted by its manufacturer for the delivery of its volume and for
the standard reference temperature of using grade 3 water as defined in ISO 3696.
If piston burettes are designed to have their factory-preset adjustment altered by the user, the
design shall prevent unintentional readjustment. If the user readjusts the piston burette, it
shall be clearly and unequivocally indicated on the outside of the piston burette that
readjustment has been effected, for example by means of a mark, a broken seal, or a label
affixed by the user.
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Type testing of the adjustment shall satisfy the performance requirements specified in clause
6 when the burette is tested in accordance with ISO 8655-6.
6.3 Transfer of hand warmth

The constructional design of piston pipettes and the materials used for their manufacture
shall be chosen in such a way that any heat transmitted from the user's hand to the apparatus
during periods of use or of test in accordance with ISO 8655-6 may be ignored.
NOTE Transfer of hand warmth will appear as a systematic drift of results during the series of tests
to ISO 8655-6.
6.4 Pipette tips
6.4.1 General
 The dispensing orifice of the tip shall be shaped in such a way that consistent dispensing of
the liquid to be measured is ascertained. When the tip is touched against the wall of a vessel
in successive operations, any amount of liquid remaining in or around the dispensing orifice
of the tip shall be consistent.
 In the case of sterilizable pipette tips the sterilization procedures indicated as appropriate by
the manufacturer in user information or on packaging shall not negatively affect the
metrological characteristics of the tips such as shape, seal and wetability.
6.4.2 Air-displacement pipette tips
 Air-displacement pipette tips shall be disposable parts, usually made of plastic, which fit on
the tip holder of the pipette and prevent the instrument from contact with the aspirated
liquid.
 Tips for piston pipettes with air interface shall be fitted in accordance with the pipette
supplier's instructions to form a good seal between the tip and the tip cone of the piston
pipette.
NOTE Variability of amount of externally retained liquid or an incomplete seal will contribute to
poor precision when testing to ISO 8655-6.
 Pipette tips made of plastic for piston pipettes with air interface are designed for single use.
They shall not be cleaned for reuse as their metrological characteristics will no longer be
reliable.
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The form of the pipette tips to be used with a multi-channel piston pipette shall be so straight that
all tips fitted are positioned with parallel axes in the same plane in order to allow for even liquid
dispensing in the target vessels, e.g. the adjacent wells of a micro liter plate. The bottoms of
properly fitted tips shall not vary in spacing from their nominal axes, nor from their common plane
by more than ± 1 mm for nominal volumes up to 350 µl and ± 1.5 mm for nominal volumes
exceeding 350 µl.
6.4.3 Positive-displacement pipette tips

 Positive-displacement pipette tips shall consist of a plunger and a capillary which fit on the
tip holder of the pipette. Various materials may be used for the plunger, such as metal or
plastic, and the capillary, such as plastic or glass. These pipette tips may be reusable or
disposable (both plunger and capillary is changed at each sampling).
 The shape and material of the plunger and capillary shall confer a good seal of the tip, as
well as a smooth action between the plunger and the capillary, to ensure consistent
dispensing of the liquid.
 Sterilizability shall not negatively affect the metrological characteristics of the tips such as
shape, seal and wetability.
7- Marking
7.1 Piston pipettes
The following data shall be permanently marked on each piston pipette:
 Nominal volume or, for variable-volume piston pipettes, useful volume range;
 Unit of measurement, e.g. l or ml;
 Supplier's name and/or trademark;
 Supplier's apparatus name or type;
 Apparatus identification serial number or an equivalent unique identifier.
In addition, the following information should be marked:
 Abbreviation “Ex” and the reference temperature 20oC;
 Reference to this part of ISO 8655, i.e. “ISO 8655-2”.
7.2 Pipette tips
The following information shall be printed on the packaging of each saleable unit of pipette tips:
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 Supplier's name and/or trademark;

 Maximum volume to be dispensed with the tip;
 Statement of whether or not the tips are suitable for sterilization.
Piston burettes
The following data shall be permanently marked on each piston burette:
 nominal volume; where this is not practicable due to a modular design, the volumetric
modules shall be marked either with their nominal volumes or information to enable the user to
obtain the nominal volume from the user information
 supplier's name and/or trademark;
 supplier's apparatus name;
 unit of measurement;
 serial number of the piston burette.
The following information should preferably also be permanently marked:
 abbreviation “Ex” and the reference temperature “ ”;
 Reference to ISO 8655-3.
Where appropriate, a conformity mark may be affixed to the piston burette
Change-over units (if applicable)
The following information shall be permanently printed or labeled on each change-over unit:
 supplier's name and/or trademark;
 nominal volume [see 8.1 a)];
 drive for which the change-over unit is suited, including indication of the supplier and
device designation of the
supplier (alternatively this information can be given in the accompanying documentation or in
the accompanying declaration of conformity);
 serial number.
8 Apparatus
8.1 Analytical balance or equivalent weighing device,
Microbalance XPE 26 PC, SN: b609161507
Comparator Balance XPE 505 C, S/n.: b609159905
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Sartorius analytical Balance te214S, S/N: 25510357

with a resolution appropriate to the selected volume of the apparatus under test (see Table 1).
Table 1 - Minimum requirements for balances

Selected Volume under Testa Resolution Standard deviation Linearity
(Repeatability)
V
mg mg mg
1 l ≤ V ≤ 10 l 0.001 0.002 0.002
10 l ≤ V ≤ 100 l 0.01 0.02 0.02
100 l ≤ V ≤ 1000 l 0.1 0.2 0.2
1 ml ≤ V ≤ 10 ml 0.1 0.2 0.2
10 ml ≤ V ≤ 200 ml 1 2 2
a
For Practical Purposes, the nominal volume may be used to choose the balance
If the standard uncertainty of measurement of the balance is known (e.g. from the balance
calibration certificate), this may be used instead of the repeatability and linearity. The standard
uncertainty of measurement shall not be more than two to three times the resolution.
8.2 Liquid reservoir
it used with sufficient capacity for all the test liquid likely to be required for the complete
series of tests.
8.3 Weighing vessel
Suitable for the test procedure selected from clause 7. Care shall be taken regarding the loss
of water by evaporation during the dispensing and weighing procedure.It is recommended
that, especially for testing apparatus of the lowest volume, the height-to-diameter ratio of the
weighing vessel be at least 3:1 or that a weighing vessel with a lid be used.
8.4 Timing device, with a standard uncertainty of 1s.
8.5 Thermometer, with a standard uncertainty of 0. 2 oC.
8.6 Hygrometer, with a standard uncertainty of 10 %.
8.7 Barometer, with a standard uncertainty of 0.5 kPa.
NOTE: All uncertainties are specified using a coverage factor of 1.
8.8 Test liquid
Use distilled or deionized water conforming grade 3 as specified in ISO 3696, degassed or air-
equilibrated. The test water shall have stood in the room for a sufficient time, at least, to reach
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equilibrium with the room conditions.
9 Test conditions
9.1 General
Apparatus that is routinely dismantled and reassembled within the scope of its intended use (e.g. for
cleaning purposes) shall be dismantled and reassembled at least once prior to the test in accordance
with the supplier's operation manual.
9.2 Test room
The test shall be carried out in a draught-free room with a stable environment. The test room shall
have a relative humidity above 50 % and a constant (± 0.5 oC ) temperature between 15 oC and 30
o
C . Prior to the test, the apparatus to be tested and the test water shall have stood in the room for a
sufficient time, at least 2 h, to reach equilibrium with the room conditions.
9.3 Evaporation
Especially for small volumes below 50 l , errors due to evaporation of the test liquid during
weighing shall be taken into consideration. Apart from the design of the weighing vessel , the test
cycle time is important. In order to keep the error due to evaporation as small as possible, the
following additional items can be considered, if volumes below 50 l are tested:
 a balance with appropriate accessories such as an evaporation trap could be used; or
 the test liquid to be weighed could be delivered into a capillary tube, although this method
does not replicate the normal method of use and the user should verify for himself that
correlation exists.
Regardless of these items, the error due to evaporation during the measuring series can be
determined experimentally and compensated mathematically. The uncertainty of this compensation
should be added to the uncertainty of measurement.
9.4 Test cycle time
The test cycle time (time required to complete the weighing of one dispensed volume) shall be kept
to a minimum. It should ideally not exceed 60 s. It is important that it is regular, both within each
cycle and as far as possible from cycle to cycle, so that a reliable mathematical compensation of the
error due to evaporation during the measuring series can be applied.
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10 Gravimetric Procedure for calibration
10.1 General
10.1.1 Test volume
In the case of a fixed-volume apparatus, the test volume is the nominal volume. In the case of a
variable-volume (user-selectable volume) apparatus, at least three volumes shall be tested:
 the nominal volume,
 approximately 50 % of the nominal volume,
 the lower limit of the useful volume range or 10 % of the nominal volume (whichever is the
greater).
Measurement of further volumes is optional. The setting devices of the apparatus (e.g. dials, scales)
shall be sufficient for the selection of the test volume.
10.1.2 Number of measurements per test volume

If the gravimetric methods are used as conformity tests or type tests, or if the gravimetric method is
used as a reference method, 10 measurements for each test volume shall be carried out. These
measurements are used to calculate the systematic and the random error of measurement. For re-
establishing conformity, e.g. after repairs not performed by the supplier, 10 measurements at each
volume shall also be performed. If the gravimetric method is used for other purposes, such as
supplier's quality control or supplier's after-sales service,
 the number of test volumes (see 10.1.1),
 the number of measurements per volume and,
 where applicable, the number of channels tested
may be changed to an appropriate number. Alternative test methods may also be used for this
purpose, provided that they can be proven to correlate with the reference method specified in ISO
8655-6, in which case the user should choose a number of measurements for his metrological
confirmation based on his accuracy requirements.
10.1.3 Weighing procedure
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Weighing for apparatus designed to deliver (Ex) shall always involve dispensing of the test liquid
into the weighing vessel. Weighing for apparatus designed to contain (In) shall always involve the
removal of test liquid from the weighing vessel.
10.1.4 Test conditions during the weighing procedure

At the start and at the end of the weighing procedure, the temperature of the test liquid in its
container shall be recorded to the nearest 0.2 oC. The barometric pressure in the test room shall be
recorded to the nearest 1 kPa and the relative humidity to the nearest 10%.
10.2 Single-channel piston pipettes with air interface (in accordance with ISO 8655-2)
10.2.1
In the case of power-driven piston pipettes, aspiration and delivery of test liquid are
automatic. The remainder of the procedure is carried out following the procedure described below.
10.2.2
Place test liquid from the water container in the weighing vessel to a depth of at least 3 mm.
Record the temperature of the test liquid and the barometric pressure and relative humidity in the
test room. If the weighing vessel has a lid, fit it.
NOTE: Temperature and barometric pressure are necessary for the choice of the correction factor Z. the
relative humidity is not necessary for the evaluation as the correction factor Z in annex A apply to relative
humilities ranging from 20 % to 90 % but are necessary for documentation in the test report.
10.2.3
If using a variable-volume piston pipette, select the test volume; this setting shall not be
altered during the test cycle of 10 measurements.
10.2.4
Prepare the piston pipette and the test cycle as follows:
 Fit the selected tip to the piston pipette.
 Fill the tip with test liquid and expel to waste five times to reach humidity equilibrium in
the dead air volume of the air-displacement piston pipette.
 Place the weighing vessel with its added water on the balance pan.
10.2.5
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Perform the following test cycle (see Figure 1 and Figure 2):
 Replace the disposable tip of the piston pipette
 Fill the piston pipette with test liquid, immersing its delivery orifice 2mm to 3mm below
the surface of the water.
 Release the operating button slowly, if hand operated, and withdraw the pipette vertically
and carefully from the surface of the water.
 Touch the delivery orifice against the side wall of the container with the test liquid.
 Expel the water to waste in order to pre-wet the tip and refill the piston pipette as
described before.
 Record the mass mo of the weighing vessel to the nearest readable graduation as in Table
1, or tare the balance to zero (mo = 0).
 Start the timing device (this may be omitted if using a weighing vessel with lid).
 If the weighing vessel has a lid, remove it. Deliver the contents of the pipette into the
weighing vessel, touching the delivery end of the pipette tip against the inside wall of the
vessel just above the liquid surface at an angle of approximately 30o to 45o and draw it
approximately to 8 mm along the inner wall of the weighing vessel to remove any droplets
at or around the tip orifice. Replace the lid if applicable.
Where applicable, use the blow-out feature of the piston pipette to expel the last drop of liquid
before drawing the delivery end of the tip along the inner wall of the weighing vessel.
If it is necessary to remove the weighing vessel from the balance pan to permit delivery of the
dispensed volume, avoid excessive handling and possible contamination by the use of lint-free
gloves. Return the weighing vessel to the balance pan after delivery.
Record the mass m1of the weighing vessel, or if tared the mass mi of the quantity delivered.
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Figure 1 Pipetting of testing volume into the weighing vessel
Figure 2 Scheme of test procedure for piston pipettes with air interface
10.2.6
Repeat the test cycle as described in 10.2.5 until 10 measurements have been recorded as a series of
masses from m1 to m10.
10.2.7
Note the time to the nearest second taken to complete the 10 test cycles.
10.2.8
After the last weighing of 10.2.6 leave the weighing vessel on the balance pan for the time
measured in 10.2.7 and record its mass. If the weighing vessel was removed from the balance pan to
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enable delivery, leave it on the pan for half the time in 10.2.7 and then remove it from the balance
and allow it to stand on the workbench for half the time measured in 10.2.7.
If the test volume is > 50 l or if a weighing vessel with lid is used, omit steps 10.2.7 and 10.2.8, as
a correction for evaporation is unnecessary. At 50 l and below, calculate the mass loss in
accordance with the supplier's instructions.
10.2.9
Measure the temperature of the remaining test liquid to the nearest 0.2 oC and calculate and
record the mean test temperature (see 10.1.4).
10.2.10
The values obtained shall be evaluated in accordance with clause 8.
10.3 Multi-channel piston pipettes (in accordance with ISO 8655-2)

Multi-channel piston pipettes are similar to single-channel in that they comprise a set of single-
volume measuring and delivery units all operated simultaneously by a single operating mechanism.
For the purposes of the test, each channel shall be regarded as a single channel and tested and
reported as such. Fill all channels of the multi-channel pipette by aspirating the test liquid. Expel
only the test liquid aspirated by the channel being tested into the weighing vessel.
10.4 Positive-displacement pipettes (in accordance with ISO 8655-2)
Piston pipettes without an air interface shall be tested in accordance with 10.2. However, the five-
fold prewetting of the pipette tip prior to the test and the single prewetting before each measurement
only need be performed if required by the supplier. Only change the pipette tips when testing
positive-displacement pipettes of type D2 (see ISO 8655-2). Wipe the pipette tip on the water
container wall after aspiration of the test liquid and prior to expelling it into the weighing vessel in
order to remove droplets from the outside of the tip. Without removing liquid from the inside of the
pipette tip, further remove any droplets that may still be present after the wiping of the tip. Follow
the supplier's instructions regarding air-bubble-free filling of the pipette tip. Empty the contents of
the pipette tip into the weighing vessel as specified in 10.2.
11 General Calculations
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It shall be applied in accordance with Annex B
12 Calculation of a Measurement Uncertainty

It shall be applied in accordance with Annex C.
13 Associated Documents
10.1 Calibration Data Sheet (……/……/……..)
10.2 Calibration Sticker (……/……/……..)
Calibration sheet of Automatic Pipette
Customer: Date:
Description: Cert. No.:
S.N.: Code: In :
Accuracy: ± Class: Manufacture:
Room temperature (oC): Humidity (%): Pressure (mbar):
Water temperature (oC):
Mass 1 (g) Mass 2 (g) Mass 3 (g)
Empty
Loaded capacity ( ml)
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Calibrated by: Reviewed by:
‫اﻟﺟﮭﺎز اﻟﻣرﻛزى ﻟﻠﺗﻘﯾﯾس و اﻟﺳﯾطرة اﻟﻧوﻋﯾﺔ‬ (COSCO)

‫رﻗم اﻟﺷﮭﺎدة‬
‫ﺗﺎرﯾﺦ اﻟﻣﻌﺎﯾرة‬
‫رﻗم اﻟﺗﺳﻠﺳل‬
‫ﺗﺎرﯾﺦ اﻋﺎدة اﻟﻣﻌﺎﯾرة‬
‫اﻟﺗوﻗﯾﻊ‬
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Annex A
(Normative)
Calculation of volume
11.1 General Calculations
 Calculate the mass of water Contain for each cycle by subtracting the balance reading
recorded at the end of the previous cycle from the reading recorded at the end of the cycle.
 To convert each mass value to a volume at 20 °C (the standard reference temperature for the
calibration of Flask) it must be divided by the density of water corrected to 20 °C. A
correction for air buoyancy, which varies with the air density, must also be applied. Both
these factors are taken into account by multiplying the each mass value by a correction factor
Z.
A table specifying the value of Z for a temperatures range of 15 °C and 30 °C and air pressure
range of 80 kPa to 105 kPa. The correction factor is also given as an equation in ISO/TR
20461:2000.
The volume V is calculated according to the following
   A 
V20  I L  I E      1   t  20 .............B1
1
  1 
 W   A    B 
or its equal to
m b   a
V20    1   c t d  t d 20 .................( B1)
 b W   a
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Where
IL is the balance reading of vessel with water, in grams;
IE is the balance reading of empty vessel, in grams (zero in case the balance was tared with the
Volumetric instrument or receiving vessel);
A is the density of air, in grams per milliliter, obtained from Table B.3 at the temperature and
atmospheric pressure of the test;
B, b is either the actual density of the balance weights when these are adjusted to their nominal
mass, or the reference density for which the weights have been adjusted (see the note
below), in grams per milliliter, or, when using an electronic balance without weights, the
(reference) density of the weights with which it has been adjusted;
NOTE
Weights conforming to International Document OIML D 28 of the International
Organization of Legal Metrology have been adjusted to give correct results when weighing
in air as though the density of the weights were 8,0 g/ml. Electronic balances are usually
adjusted by means of these weights.
W is the density of water at t °C, in grams per milliliter, calculated with the “Tanaka” formula
(see Table B.4 in);
γ,α is the coefficient of cubical thermal expansion of the material of which the volumetric
instrument tested is made, in reciprocal degrees Celsius (see Table B.5);
t is the temperature of the water used in testing, in degrees Celsius.
Tables B.6, B.7 and B.8 give factor Z conversion values for different types of glass at
common air Pressure versus temperature. In these tables, the combined effects of the density
of the water.
td Temperature of the glassware (oC)
td20 Reference temperature of the glassware (oC)
The thermal expansion of the glass and the air buoyancy has been taken into account. The
used balance weight is ρB = 8,0 g/ml.
When the temperature at which the volumetric instrument is used (t2) differs from the reference
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temperature (t1), the volume of the volumetric instrument at (t2) can be calculated from the
following equation:
……………………….. B.2
where
γ is the coefficient of cubical thermal expansion (see Table B.5). For information on the effect
of temperature differences.
11.2 Tables for calculation
To facilitate an easy calculation of the instrument's volume V at a reference temperature of

20 °C from the apparent mass obtained by using a balance, a factor Z can be introduced in
Equation (B.1):
…………..….. B.3
common air pressure versus temperature. In these tables, the combined effects of the density
of the water, the thermal expansion of the glass and the air buoyancy have been taken into
account. The used balance weight is B = 8, 0 g/ml.
The factor Z conversion values have been derived from Equation (B.1) as follows:
   A 
  1   t  20 .............B.4 
1
Z     1 
 W   A    B 
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Annex B
(Normative)
Calculation of a Measurement Uncertainty
Determination of uncertainty is calculated according to the Technical Report ISO/TR

20461:2000 – edition 1 together with a correction note ISO/TR 20461:2000
1. A conventional volume of a calibrated pipette in temperature 20 ºC is determined by the

following formula:
(1)
,
(2)
where: m is the balance reading of delivered water;

m1 is the balance reading of the weighing vessel before delivery of the measured
volume of water;
m2 is the balance reading of the weighing vessel after delivery of the measured
volume of water;
mE is the balance reading of the mass loss due to evaporation of liquid during the
measurement;
Z is the combined factor for buoyancy correction and conversion from mass to
volume;
Y is the thermal expansion correction factor of the delivering device.
Z is given by
(3)
where w is the density of water;

a is the density of air;
b is the density of the standard weight used to calibrate the balance
[according to OIML
b = 8 000 kg/m3for steel weights].
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(4)
where tw is the water temperature in degrees Celsius;

with the constants (ITS-90 temperature scale):
a0 is equal to 999,853 08 kg/m3
a1 is equal to is 6,326 93×10–2 °C–1 kg/m3 ;
a4 is equal to is 3,821 216×10–7 °C–4 kg/m3.
(5)
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(6)
From the last equations one may write:
(7)
(8)
9.1 Standard uncertainty of measurement associated with the volume V20
According to the GUM the standard uncertainty of measurement associated with the value V20 may
be written as:
(9)
(10
)
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The sensitivity coefficients may be determined by calculating the partial derivatives as indicated in
the equation, by numerical calculations, or by experiment. As the uncertainties of the constants
[equation (8)] and the uncertainties of equations (4) and (5) for a and w very small compared to
other uncertainties, they may be neglected in the evaluation of uncertainty.
12.2 Sensitivity coefficients
The evaluation of the uncertainty of measurement does not require such exact values and exact
solutions of the mathematical model for the measurement, as the determination of the volume V20
itself. Approximations are tolerable, but they have to be used only for this uncertainty evaluation.
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12.3 Expanded uncertainty of measurement associated with volume V20
The expanded uncertainty of the volume V20 is expressed as:
(32)
(33)
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12.4 The final Uncertainty Budget
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Procedure for calibration Identification:
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(COSQC)- Metrology Department YYYY-MM-DD
WORK PROCEDURE
FOR
CALIBRATION OF PIPETTES and BURETTES
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1 Purpose 3
2 Scope 3
5 General Requirements 4
6 Apparatus and calibration liquid 6
7 Gravimetric Procedure for Calibration 7
8 Calculations 7
11 APPENDIX A 11
12 APPENDIX B 13
13 APPENDIX C 24
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1.0 Purpose
Purpose of this Lab Calibration Procedure is to provide guideline for the calibration of Burette
and Pipette to Deliver.
2.0 Scope
 Calibration of Burette. From 1ml to 50ml:

 Calibration of Pipette. From 10ml to 100ml
 ISO 4787 Laboratory glassware - Volumetric instruments - Methods for testing of

capacity and for use
 Operating Manual of Mettler balance ( xp505c,).
 Operating Manual of Sartorius balance (TE214S).
4.1 Calibration
The set of operations which establishes under specific conditions the relationship between
value indicated by a measuring instrument or measuring system or values represented by a
material measure and the corresponding known value of the measurand.
4.2 Traceability
4.6 Uncertainty of Measurement
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It is an estimate of the uncertainty of the result of a calibration. The estimate is obtained by

multiplying the combined uncertainty by a coverage factor.
4.9 Repeatability
4.10 Resolution
5.0 General Requirements

5.1 Laboratory Environment
The test shall be carried out in a draught-free room with stable environment. The test room
shall have a relative humidity between 35 % and 85 % and shall provide a temperature locally
constant to ±1 °C and temporally constant to ±0,5 °C between 15 °C and 30 °C. Prior to the
test, the volumetric instrument to be tested and the test water shall have stood in the room for
a sufficient time (1 h to 2 h) to reach equilibrium with the room conditions. Test water should
be covered to avoid evaporation cooling. Temperatures (room and calibration liquid),
atmospheric pressure and humidity should be recorded.
5.2 Temperature of calibration liquid

The temperature of the water used for the calibration shall be measured to ± 0.1 °C.
Corrections for differences in temperature from the reference temperature shall be applied in
accordance with Annex B.
5.3 Cleanliness of glass surface

It shall be applied in accordance with Annex A.
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5.4 Quality of used volumetric instruments
The glass surface shall be free from obvious damage, the graduations and inscriptions shall
be clearly readable and especially with instruments adjusted to deliver the jet shall be free from
damage and allow an unrestricted outflow of liquid.
5.5 Setting the meniscus
Most volumetric instruments employ the principle of setting or reading a meniscus (the
interface between air and the liquid) against a graduation line or ring mark. Wherever practicable,
the meniscus should descend to the position of setting. The tubing of the volumetric instrument
shall be in a vertical position. The eye of the testing person shall be in the same horizontal plane as
the meniscus or the graduation line (ring mark).
Key Key
1 meniscus of liquid 1 meniscus
2 graduation line or ring mark 2 graduation line
3 blue or black (dark) paper or black rubber tubing 3 Schellbach ribbon
Figure 1 — Setting of the meniscus with Figure 2 — Meniscus with Schellbach ribbon
transparent liquids
6 Apparatus and calibration liquid
6.1 Weighing Equipment
Performance of gravimetric calibration procedure of glassware requires use of a weighing

instrument with metrological parameters as described in ISO 4787.
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Comparator Balance XPE 505 C, S/n.: b609159905,

Sartorius analytical Balance te214S, S/N: 25510357,

(Repeatability)
V
mg mg mg
100 l ≤ V ≤ 10 ml 0.1 0.2 0.2
10 ml ≤ V ≤ 1000 ml 1 1 2
1000 ml ≤ V ≤ 2000 ml 10 10 20
V > 2000 ml 100 100 200
a
In case, where standard deviation of liquid measurement is specified, than it is possible to

use it as acceptance criterion instead of repeatability and linearity parameters of a weighing
instrument. However, it needs to be assumed, that standard uncertainty is not bigger than
twofold or threefold of reading unit d.
In order to maintain measurement re-traceability, a weighing instrument that is utilized for
glassware calibration, should have valid calibration certificate.
6.2 Thermometer
to measure the temperature of the calibration liquid (water) with a measurement error of
maximum 0,2 °C for liquid volumes < 1 000 ml and with a measurement error of maximum
0,1 °C for liquid volumes W 1 000 ml.
6.3 Hygrometer
to measure the humidity in the test room with a measurement error of maximum 5 % within
the humidity range of 35 % to 85 %.
6.4 Barometer
to measure the atmospheric pressure in the test room with a measurement error of
maximum 1 kPa.
6.5 Calibration liquid
Distilled or deionized water complying with ISO 3696, Grade 3 should be used for testing.
6.6 Receiving vessel
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Conical flask with ground joint, manufactured from glass, e.g. in accordance with ISO
4797. The nominal volume of the conical flask shall correspond to the volume of liquid to
be measured.
7.1 For Pipettes (One Mark) To Deliver
1- Fill the pipette with distilled water by suction to the index mark and measure the delivery
time with the tip in contact with the glass surface of the internal side of a beaker. Refill by
suction slightly above the index line. Record the temperature of the distilled water in the
beaker from which the pipette is filled.
Remove any water on the outside of the tip by a downward wipe with filter paper after the
filling is completed.
2- Then slowly lower the meniscus to the index using either a stopcock or hose clamp for "fine
control." The tip must be in contact with the wet wall of a beaker while the setting is made
on the index line.
3- Do not remove any water remaining on the tip at this time. Hold the pipette in a vertical
position and deliver water into a previously weighed weighing flask with the tip in contact
with the inside wall of the neck of the flask. After the water has ceased to flow, wait 2 s,
then remove the pipette from contact with the flask. The flask now weighed with its
contained load.
7.2 For Burettes to Deliver
1- Clamp the burette vertically on a support stand.

2- Clamp a plain glass test tube, large enough to hold a thermometer, near the burette if the
burette is of such a size that it is not large enough to insert a thermometer in the top for
observing the temperature of the water.
3- Fill the burette from a reservoir or storage bottle, in which the water has reached equilibrium
with room temperature, and check to verify that there is neither leakage from the tip nor
from the stopcock plug.
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4- Drain and record the delivery time.

5- Refill the burette to approximately 10 mm above the zero mark and fill the test tube that
holds the thermometer; record the temperature. Set the meniscus on the zero mark using the
burette stopcock to lower the liquid level and touch the tip with the wetted wall of a beaker
to remove any excess water.
6- A weighing flask that has been cleaned and , weighed empty is placed with the inside
of the neck in contact with the tip of the burette (the flask will be at a slight angle).
7- Fully open the stopcock until the water is only a few millimeters above the line being tested
and then the stream is slowed so as to make an accurate setting. When the setting has been
completed, move the flask horizontally. Breaking the contact with the burette.
8- Recheck the setting then stopper and weigh the flask a second time, after which refill the
thermometer tube and test the next interval in the same manner as the first one-from the zero
mark to the next interval needed.
9- For burettes with a specified waiting time, use the following procedure: after adjustment to
the zero mark, fully open the stopcock until the meniscus has reached a position a few
millimeters above the graduation line for calibration. After the specified waiting time (for
example, 30 s), adjust the meniscus to the graduation line, remove the flask, and weigh.


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Calibration sheet of Pipette / Burette
Customer: Date:
S.N.: Code: In :
1 Mass 1 (g) Mass 2 (g) Mass 3 (g)
Empty
Empty
Empty
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Annex A
(Informative)
Cleaning of volumetric glassware
Cleanliness of glass surface
The volume contained in, or delivered by, a volumetric instrument depends on the cleanliness
of the internal glass surface. Lack of cleanliness results in errors through a poorly shaped
meniscus involving two defects:
 incomplete wetting of the glass surface, i.e. the liquid surface meets the glass at an arbitrary
angle instead of forming a curve such that it meets the glass tangentially;
 a generally increased radius of curvature, due to contamination of the liquid surface reducing
the surface tension.
The ascending or descending liquid meniscus shall not change shape (i.e. it shall not crinkle at
its edges). To ascertain whether a piece of glass apparatus is satisfactorily clean, it shall be
observed during filling and dispensing. Additionally, an experienced operator can recognize
the shape of an uncontaminated meniscus, in relation to its diameter.
Lack of cleanliness causes additional errors with volumetric instruments used for delivery due
to the film of liquid on the walls being irregularly distributed or incomplete, e.g. forming
drops on the glass surface. Furthermore, chemical residues can introduce an error in the
analytical result by contamination. Therefore, where volumetric instruments are fitted with
ground stoppers, special attention shall be paid to cleaning the ground zone.
 Glassware can be cleaned manually, in an immersion bath or in a laboratory washing

machine. To reduce volume changes through glass erosion and destruction of graduations,
gentle cleaning with detergents of low alkalinity at temperatures below 70 °C with short
contact time and whenever possible immediately after use is recommended.
 If the inner glass walls are not sufficiently clean after the above treatment, the volumetric
instrument should be filled with a mixture of equal parts of a 30 g/l solution of potassium
permanganate (KMnO) and 1 mol/l solution of sodium hydroxide (NaOH). After about 2 h, a
residue of MnO may be removed by means of dilute hydrochloric acid or oxalic acid. The
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volumetric instrument should then be rinsed with distilled water and it should again be
ascertained that the walls are sufficiently clean. If they are not, the procedure should be
repeated. If this treatment is not successful, specific cleaning methods described in laboratory
handbooks should be applied. The method shall not change the volume of the instrument.
As a safeguard, it is recommended that volumetric instruments should not be heated to a
temperature considerably above 180 °C. Although the strain point of glasses used for volumetric
purposes is in the range of 500 °C, alterations of volume might occur at temperatures considerably
below the strain point.
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Annex B
(Normative)
a) Calculate the mass of water Contain for each cycle by subtracting the balance reading
b) To convert each mass value to a volume at 20 °C (the standard reference temperature for the
Z.
20461:2000.
   A 
V20  I L  I E      1   t  20 .............B1
1
  1 
 W   A    B 
or its equal to
m b   a
V20    1   c t d  t d 20 .................( B1)
 b W   a
Where
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NOTE
(see Table B.4 in);
of the water.
following equation:
……………………….. B.2
where

Equation (B.1):
…………..….. B.3
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   A 
  1   t  20 .............B.4 
1
Z     1 

 W   A    B 
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Annex C
(Normative)


following formula:
(1)
,
(2)

volume of water;
volume of water;
measurement;
volume;
Z is given by
(3)

[according to OIML
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(4)

(5)
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(6)
(7)
(8)
be written as:
(9)
(10
)
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(32)
(33)
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WORK PROCEDURE
FOR
CALIBRATION OF Flasks and Cylinders
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1 Purpose 3
2 Scope 3
5 General Requirements 4
6 Apparatus and calibration liquid 6
7 Gravimetric Procedure for Calibration 7
8 Calculations 7
11 APPENDIX A 11
12 APPENDIX B 13
13 APPENDIX C 24
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1.0 Purpose
Purpose of this Lab Calibration Procedure is to provide guideline for the calibration of Flask
and Cylinder to Contain.
2.0 Scope
 Calibration of Flask. From 5ml to 2000 ml

 Calibration of Cylinder. 10ml to 2000 ml
 ISO 4787 Laboratory glassware - Volumetric instruments - Methods for testing of

capacity and for use
 Operating Manual of Mettler balance ( xp505c,xp10001c, 32kg).
 Operating Manual of Sartorius balance (TE214S).
 Operating Manual of Ohaus balance (TE214S).
4.1 Calibration
The set of operations which establishes under specific conditions the relationship between
value indicated by a measuring instrument or measuring system or values represented by a
material measure and the corresponding known value of the measurand.
4.2 Traceability
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4.6 Uncertainty of Measurement

It is an estimate of the uncertainty of the result of a calibration. The estimate is obtained by
multiplying the combined uncertainty by a coverage factor.
4.9 Repeatability
4.10 Resolution
4.12 Dispense
To deliver the sample.
5.0 General Requirements

5.1 Laboratory Environment
The test shall be carried out in a draught-free room with stable environment. The test room
shall have a relative humidity between 35 % and 85 % and shall provide a temperature locally
constant to ±1 °C and temporally constant to ±0,5 °C between 15 °C and 30 °C. Prior to the
test, the volumetric instrument to be tested and the test water shall have stood in the room for
a sufficient time (1 h to 2 h) to reach equilibrium with the room conditions. Test water should
be covered to avoid evaporation cooling. Temperatures (room and calibration liquid),
atmospheric pressure and humidity should be recorded.
5.2 Temperature of calibration liquid

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The temperature of the water used for the calibration shall be measured to ± 0.1 °C.
Corrections for differences in temperature from the reference temperature shall be applied in
accordance with Annex B.
5.3 Cleanliness of glass surface

It shall be applied in accordance with Annex A.
5.4 Quality of used volumetric instruments
The glass surface shall be free from obvious damage, the graduations and inscriptions shall
be clearly readable and especially with instruments adjusted to deliver the jet shall be free from
damage and allow an unrestricted outflow of liquid.
5.5 Setting the meniscus
Most volumetric instruments employ the principle of setting or reading a meniscus (the
interface between air and the liquid) against a graduation line or ring mark. Wherever practicable,
the meniscus should descend to the position of setting. The tubing of the volumetric instrument
shall be in a vertical position. The eye of the testing person shall be in the same horizontal plane as
the meniscus or the graduation line (ring mark).
Key Key
1 meniscus of liquid 1 meniscus
2 graduation line or ring mark 2 graduation line
3 blue or black (dark) paper or black rubber tubing 3 Schellbach ribbon
Figure 1 — Setting of the meniscus with Figure 2 — Meniscus with Schellbach ribbon
transparent liquids
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6 Apparatus and calibration liquid
6.1 Weighing Equipment
Performance of gravimetric calibration procedure of glassware requires use of a weighing

instrument with metrological parameters as described in ISO 4787.
Comparator Balance XPE 505 C, S/n.: b609159905,
Sartorius analytical Balance te214S, S/N: 25510357,
Ohaus analytical Balance ARA 520, s/n: 1228380181,
Mettler Toledo Digital Balance, SG32001, S/N: 1121320019

(Repeatability)
V
mg mg mg
100 l ≤ V ≤ 10 ml 0.1 0.2 0.2
10 ml ≤ V ≤ 1000 ml 1 1 2
1000 ml ≤ V ≤ 2000 ml 10 10 20
V > 2000 ml 100 100 200
a
In case, where standard deviation of liquid measurement is specified, than it is possible to

use it as acceptance criterion instead of repeatability and linearity parameters of a weighing
instrument. However, it needs to be assumed, that standard uncertainty is not bigger than
twofold or threefold of reading unit d.
In order to maintain measurement re-traceability, a weighing instrument that is utilized for

glassware calibration, should have valid calibration certificate.
6.2 Thermometer
to measure the temperature of the calibration liquid (water) with a measurement error of
maximum 0,2 °C for liquid volumes < 1 000 ml and with a measurement error of maximum
0,1 °C for liquid volumes W 1 000 ml.
6.3 Hygrometer
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to measure the humidity in the test room with a measurement error of maximum 5 % within
the humidity range of 35 % to 85 %.
6.4 Barometer
to measure the atmospheric pressure in the test room with a measurement error of
maximum 1 kPa.
6.5 Calibration liquid
Distilled or deionized water complying with ISO 3696, Grade 3 should be used for testing.
6.6 Receiving vessel
Conical flask with ground joint, manufactured from glass, e.g. in accordance with ISO
4797. The nominal volume of the conical flask shall correspond to the volume of liquid to
be measured.
7.1 For Flask to Contain
1- After cleaning and drying, weigh the empty flask including the stopper.
2- Place an appropriate sized funnel in the flask to dis- charge the water below the stopper.
3- Fill from beaker or supply line, maneuvering the funnel So as to wet the entire neck
below the stopper. Let stand for about 2 min to allow the walls to drain.
4- Place the flask under a burette and complete filling and setting the meniscus, taking care
not to splash water on the walls, after which place stopper in neck to lessen evaporation
and weigh.
NOTE: A pipette or dropper with a finely drawn tip may be used to adjust the meniscus instead of
the burette.
5- Determine the temperature by placing a thermometer in the filling beaker or in one
which has been filled from the water supply. The temperature may be taken after final
weighing by placing a thermometer directly in the flask, provided the flask is of
sufficient size to accommodate it.
7.2 For Cylinder to Contain

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Using the same Procedure as Flask for calibration point


Calibration sheet of one mark flask
Customer: Date:
S.N.: Code: In :
Mass 1 (g) Mass 2 (g) Mass 3 (g)
Empty
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Calibration sheet of A cylinder
Customer: Date:
S.N.: Code: In :
Empty
Empty
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Empty

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Annex A
(Informative)
Cleaning of volumetric glassware
Cleanliness of glass surface
The volume contained in, or delivered by, a volumetric instrument depends on the cleanliness
of the internal glass surface. Lack of cleanliness results in errors through a poorly shaped
meniscus involving two defects:
 incomplete wetting of the glass surface, i.e. the liquid surface meets the glass at an arbitrary
angle instead of forming a curve such that it meets the glass tangentially;
 a generally increased radius of curvature, due to contamination of the liquid surface reducing
the surface tension.
The ascending or descending liquid meniscus shall not change shape (i.e. it shall not crinkle at
its edges). To ascertain whether a piece of glass apparatus is satisfactorily clean, it shall be
observed during filling and dispensing. Additionally, an experienced operator can recognize
the shape of an uncontaminated meniscus, in relation to its diameter.
Lack of cleanliness causes additional errors with volumetric instruments used for delivery due
to the film of liquid on the walls being irregularly distributed or incomplete, e.g. forming
drops on the glass surface. Furthermore, chemical residues can introduce an error in the
analytical result by contamination. Therefore, where volumetric instruments are fitted with
ground stoppers, special attention shall be paid to cleaning the ground zone.
 Glassware can be cleaned manually, in an immersion bath or in a laboratory washing

machine. To reduce volume changes through glass erosion and destruction of graduations,
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gentle cleaning with detergents of low alkalinity at temperatures below 70 °C with short
contact time and whenever possible immediately after use is recommended.
 If the inner glass walls are not sufficiently clean after the above treatment, the volumetric
instrument should be filled with a mixture of equal parts of a 30 g/l solution of potassium
permanganate (KMnO) and 1 mol/l solution of sodium hydroxide (NaOH). After about 2 h, a
residue of MnO may be removed by means of dilute hydrochloric acid or oxalic acid. The
volumetric instrument should then be rinsed with distilled water and it should again be
ascertained that the walls are sufficiently clean. If they are not, the procedure should be
repeated. If this treatment is not successful, specific cleaning methods described in laboratory
handbooks should be applied. The method shall not change the volume of the instrument.
As a safeguard, it is recommended that volumetric instruments should not be heated to a
temperature considerably above 180 °C. Although the strain point of glasses used for volumetric
purposes is in the range of 500 °C, alterations of volume might occur at temperatures considerably
below the strain point.
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Annex B
(Normative)
a) Calculate the mass of water Contain for each cycle by subtracting the balance reading
b) To convert each mass value to a volume at 20 °C (the standard reference temperature for the
Z.
20461:2000.
   A 
V20  I L  I E      1   t  20 .............B1
1
  1 

 W   A    B 
or its equal to
m b   a
V20    1   c t d  t d 20 .................( B1)
 b W   a
Where
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NOTE
(see Table B.4 in);
of the water.
following equation:
……………………….. B.2
where
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Equation (B.1):
…………..….. B.3
   A 
  1   t  20 .............B.4 
1
Z     1 
 W   A    B 
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Annex C
(Normative)


following formula:
(1)
,
(2)

volume of water;
volume of water;
measurement;
volume;
Z is given by
(3)

[according to OIML
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(4)

(5)
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(6)
(7)
(8)
be written as:
(9)
(10
)
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(32)
(33)
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Page 30 of 30

CALIBRATION OF Piston Pipette and Buritte PDF

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CALIBRATION OF Piston Pipette and Buritte PDF

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Procedure for calibration

CALIBRATION OF Piston Pipette and Buritte

Table of CONTENTS Page No.

 Calibration of piston pipettes (as ISO 8655-6) from 1µl to 10ml:

3.0 Normative references

 ISO 8655-1,2002 Terminology, general requirements and user recommendations

4.1 Maximum permissible error

A standard generally of a highest metrological quality available from which measurements

5.1 for Piston Pipette

5.2 For Piston Burette

Figure 2 — Schematic drawing of a piston burette

6.1 Types of piston pipette

1. For Piston Pipette

2. For Piston Burette

6.3 Transfer of hand warmth

6.4 Pipette tips

6.4.3 Positive-displacement pipette tips

 Supplier's name and/or trademark;

Sartorius analytical Balance te214S, S/N: 25510357

Table 1 - Minimum requirements for balances

equilibrium with the room conditions.

10 Gravimetric Procedure for calibration

10.1.2 Number of measurements per test volume

10.1.3 Weighing procedure

10.1.4 Test conditions during the weighing procedure

Figure 1 Pipetting of testing volume into the weighing vessel

10.3 Multi-channel piston pipettes (in accordance with ISO 8655-2)

10.4 Positive-displacement pipettes (in accordance with ISO 8655-2)

It shall be applied in accordance with Annex B

12 Calculation of a Measurement Uncertainty

Calibration sheet of Automatic Pipette

Description: Cert. No.:

Accuracy: ± Class: Manufacture:

Room temperature (oC): Humidity (%): Pressure (mbar):

Water temperature (oC):

Mass 1 (g) Mass 2 (g) Mass 3 (g)

Loaded capacity ( ml)

Calibrated by: Reviewed by:

‫اﻟﺟﮭﺎز اﻟﻣرﻛزى ﻟﻠﺗﻘﯾﯾس و اﻟﺳﯾطرة اﻟﻧوﻋﯾﺔ‬ (COSCO)

The volume V is calculated according to the following

To facilitate an easy calculation of the instrument's volume V at a reference temperature of

Determination of uncertainty is calculated according to the Technical Report ISO/TR

1. A conventional volume of a calibrated pipette in temperature 20 ºC is determined by the

where: m is the balance reading of delivered water;

where w is the density of water;

where tw is the water temperature in degrees Celsius;

From the last equations one may write:

9.1 Standard uncertainty of measurement associated with the volume V20

12.2 Sensitivity coefficients

12.3 Expanded uncertainty of measurement associated with volume V20

The expanded uncertainty of the volume V20 is expressed as:

12.4 The final Uncertainty Budget

CALIBRATION OF PIPETTES and BURETTES

Table of CONTENTS Page No.

 Calibration of Burette. From 1ml to 50ml:

3.0 Normative references

 ISO 4787 Laboratory glassware - Volumetric instruments - Methods for testing of

4.0 Terms and definitions

4.6 Uncertainty of Measurement

It is an estimate of the uncertainty of the result of a calibration. The estimate is obtained by

5.0 General Requirements