ASTM E1154-89(2008)
(Specification)Standard Specification for Piston or Plunger Operated Volumetric Apparatus
Standard Specification for Piston or Plunger Operated Volumetric Apparatus
ABSTRACT
This specification covers piston or plunger operated volumetric apparatus (POVA), in particular, the requirements, operating conditions, and test methods. POVA covered by this specification are pipettes, dispensers (with and without valve), dilutors, and displacement burets (with and without valve). Single measurement, replicate delivery, durability, functional (such as tests for leakage, broken parts, existence of air bubbles, and contamination), volumetric, and gravimetric tests shall be performed and shall conform to the requirements specified.
SCOPE
1.1 This specification covers requirements, operating conditions, and test methods for piston or plunger operated volumetric apparatus (POVA).
1.2 This specification includes specifications applicable for all types of POVA or those given by the manufacturer. The following precautionary caveat pertains only to the test method portion, Section 13, of this specification: This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
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Designation:E1154 −89(Reapproved2008)
Standard Specification for
Piston or Plunger Operated Volumetric Apparatus
This standard is issued under the fixed designation E1154; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 3.1.2.1 Discussion—The dead volume should not be con-
fused with the dead space of an air displacement instrument.
1.1 This specification covers requirements, operating
conditions, and test methods for piston or plunger operated 3.1.3 disposable—those parts of an instrument that are
volumetric apparatus (POVA). intended to be used once only and then discarded. Disposable
parts are generally intended for use in applications where
1.2 This specification includes specifications applicable for
sample carryover is intolerable.
all types of POVA or those given by the manufacturer. The
followingprecautionarycaveatpertainsonlytothetestmethod 3.1.4 maximum error—the maximum difference between
portion, Section 13, of this specification: This standard does
thenominalvolumeandanysingleindividualvolumeobtained
not purport to address all of the safety concerns, if any, by applying the test procedure specified in Section 13 of this
associated with its use. It is the responsibility of the user of this ISO Standard.
standard to establish appropriate safety and health practices
3.1.5 maximum expectable error—with more than 95%
and determine the applicability of regulatory limitations prior
probability, the maximum expectable error is calculated as
to use.
follows:
6~1E 112s! (1)
T
2. Referenced Documents
where:
2.1 ISO Documents:
E = inaccuracy of the mean, and
T
ISO3534Statistics—Vocabulary and Symbols
s = standarddeviationfromtherepeatabilitytestinSection
ISO653 Long Solid-Stem Thermometers for Precision Use
13.
ISO655 Long Enclosed-Scale Thermometers for Precision
Use 3.1.6 nominal volume(s)—the stated volume(s) for which
performance is specified.
ISO4787Laboratory Glassware—Volumetric Glassware—
Methods for Testing and Use
3.1.7 unit of volume—themillilitreorthemicrolitre,thatare
accepted substitutes for the cubic centimetre or cubic millime-
3. Terminology
tre.
3.1 Definitions of Terms Specific to This Standard: 3.1.7.1 Discussion—Itisrecommendedthatvolumesshould
3.1.1 accuracy —the accuracy of an instrument is the be specified in microlitres up to 999 µL, and in millilitres from
closeness of agreement between the nominal volume and the 1 mL.
mean volume, obtained by applying the test procedure speci-
3.1.8 piston or plunger operated volumetric apparatus
fied in Section 13 of this specification. It is quantified by the
(POVA)—the volume of liquid to be measured with POVA is
inaccuracy of the mean.
defined by one or more strokes of one or more pistons or
3.1.2 dead volume—the dead volume is that part of the total
plungers. POVA may be operated manually or mechanically
liquidvolume,heldintheoperationalpartofthedevice,which (for example, electrically, pneumatically or by hydrostatic
is not delivered.
pressure).
3.1.8.1 Discussion—In the following text the word ‘piston’
means ‘piston or plunger.’
1 3
This specification is under the jurisdiction of ASTM Committee E41 on
3.1.9 precision —the closeness of agreement between the
Laboratory Apparatusand is the direct responsibility of Subcommittee E41.06 on
individual volumes obtained by applying the test procedure
Weighing Devices.
specified in this specification. It is quantified by the impreci-
Current edition approved Nov. 1, 2008. Published January 2009. Originally
approved in 1987. Last previous edition approved in 2003 as E1154-89(2003).
sion.
DOI: 10.1520/E1154-89R08.
3.1.9.1 Discussion—The test procedure specified gives only
Available fromAmerican National Standards Institute (ANSI), 25 W. 43rd St.,
a measure of the repeatability (see ISO3534) under controlled
4th Floor, New York, NY 10036, http://www.ansi.org.
These definitions apply only in the cases where the distributions are Gaussian. conditions.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E1154−89(2008)
3.1.10 reusable—thosepartsofaninstrumentthataremeant 4.2.1 Displacement with an air interface (“air displace-
tobeusedmorethanonce.Asthereusabilityofsomepartscan ment”). The delivered liquid is displaced by an air interface
rarely be quantified, any institution or individual who reuses a (indirect action), (see Figs. 1 and 2).
reusable part must see to its safety and effectiveness. Reusable 4.2.2 Displacement without an air interface (“positive dis-
parts are generally intended for use in applications where placement”). The delivered liquid is displaced either by a
sample carryover is tolerable, or can be adequately prevented. liquid interface (indirect action) or by actual contact with the
piston (direct action), (see Fig. 3 and Fig. 4).
3.1.11 sample carryover—that portion of the sample that is
retained in the instrument and that may affect subsequent
5. Performance Requirements
samples.
5.1 Performance Tolerances:
3.1.12 stated feature—any feature claimed by the manufac-
5.1.1 Performance tolerances specified for POVAare meant
turer.
to include any thermal drift effect upon the accuracy and
3.1.13 reference temperature—the temperature at which the
precision attributable to hand-transmitted heat during normal
instrument is designed to deliver its nominal volume(s).
use. It is, therefore, important that the instrument being
3.1.13.1 Discussion—At that temperature the closest agree-
evaluated according to the referenced procedure not be pre-
ment between manufacturer’s performance claims and test
conditioned (warmed) by recent handling, nor isolated from
results may be expected.
normal handwarming during the test series (30 or 10 cycles).
3.1.14 reference temperature range—thattemperaturerange
5.1.2 Volumetricperformancetolerancesarenotspecifiedin
for which the tolerances for accuracy are specified.
this specification. The manufacturer shall specify the perfor-
¯
3.1.15 working range—that part (of the total range) for mance tolerances in terms of the accuracy of the mean (E %)
C
which manufacturer’s performance specifications are given. and coefficient of variation (CV %).Values shall be given for
c
the minimum and maximum volumes of the working range, as
3.1.16 working temperature range—that range of tempera-
wellasforanyintermediatevolumesintheseries1,2,5,10.
tures for which manufacturer’s performance specifications are
given.
5.2 The reference temperature recommended for all POVA
is 21.5°C, which is the mid-point of the reference temperature
4. Classification
range, (see section 3.1.14). The use of another reference
4.1 Types of POVA—Piston or plunger operated volumetric temperature must be stated by the manufacturer.
apparatus (POVA) are classified as follows: 5.2.1 Reference Temperature Range—The reference tem-
4.1.1 Pipette—A measuring instrument for the transfer of a perature range for all POVA shall be 19 to 24°C, (see section
predetermined volume of liquid from one vessel to another. It 3.1.13 and section 3.1.14).
is not connected to a reservoir.
5.3 Removable Parts:
4.1.2 Dispenser—A measuring instrument for delivering
5.3.1 The volumetric performance of POVAto be used with
predetermined volumes of liquid from a reservoir. The reser-
removable parts can depend to a large extent on the design,
voir may be integrated with the instrument or connected
material, and workmanship of those parts. The test procedures
externally.
described can give information only about the performance of
4.1.3 Dilutor—Ameasuring instrument for taking up differ-
the instruments together with the removable parts actually
ent liquids (for example, sample and diluent) and delivering
used.
them in combination so as to comprise a predetermined ratio,
5.3.2 Single-Measurement Test—The single-measurement
or predetermined volumes, or both. The reservoir of diluent
menttestrequireseither30or10randomlyselectedremovable
may be integrated with the instrument or connected externally.
parts, one for each sample of the series. This test evaluates the
4.1.4 Displacement Buret—A measuring instrument from
instrument’s performance and component of imprecision due
which the volume delivered is determined by an external
to the variation of these parts.
indicator. The volume delivered can then be read.
5.3.3 Replicate-Delivery Test—The replicate delivery test
4.2 Types of Displacement: uses one removable part for the 30 or 10 sample series. This
FIG. 1 Displacement With an Air Interface (Air Displacement)
E1154−89(2008)
FIG. 2 Displacement Without an Air Interface (Positive Displacement)
FIG. 3 Pipetter Mode of Operation (Forward Mode)
FIG. 4 Pipetter Mode of Operation (Reverse Mode)
testevaluatestheinstrument’sperformanceandthecomponent 6.3 Preparation—The manufacturer shall provide instruc-
of imprecision due to the reuse of this part.
tions necessary for the preparation of the instrument for use in
particular operating modes (for example, mounting of remov-
5.4 Durability—Anyclaimbyamanufacturerthataninstru-
able parts, method of volume adjustment, temperature
ment is resistant to any defined conditions (for example,
equation, isothermal requirements, testing of piston action,
sterilization and chemical exposure) shall be understood in
lubrication, priming, purging or prerinsing information, etc.).
such a way that even long term or repeated exposure to those
conditions(asspecifiedbythemanufacturer)willnotaffectthe
rated performance of the instrument. 7. Operating Conditions for Pipetters
6. General Operating Conditions
7.1 Twocommonmodesofoperationareinuse,theforward
mode (sometimes referred to as normal mode), and the reverse
6.1 Relationship to Performance—The specification of op-
mode (usable with two-component stroke mechanism systems
erating procedures is critical to the proper functioning of the
only), (see Fig. 3 and Fig. 4).
instruments, and determines their ability to perform within
specified tolerances. Changes in the operating mode can
7.1.1 In general, the precision of the repetitive use of the
dramatically alter the results of analyses. Most instruments are
forward mode relies upon the precise draining by air pressure
calibrated for certain operating modes; another manner of use
(in the case of air displacement pipetters) or internal wiping of
may result in a change in the accuracy or precision, or both.
thepipetbarrelortip(inthecaseofdisplacementpipetters).As
compared to the reverse mode, the forward mode is relatively
6.2 Delineation—It is the manufacturer’s responsibility to
insensitivetovariationsinthespeedofthepistonorplungerin
delineate the modes of operation in instruction manuals and to
state for which of the modes the instrument is calibrated. the dispensing action. Positive displacement instruments with
E1154−89(2008)
relatively small delivery orifices are generally less sensitive to 7.2.3.3 Allow the push-button to move up to the top stop
change in accuracy when handling liquids with high wetability position.
characteristics.
7.3 Reverse Mode, General Format:
7.1.2 Airdisplacementpipetterswithtwo-componentstroke
7.3.1 Preparation—Prepare in accordance with 7.2.1, for-
mechanisms are generally less sensitive than air displacement
ward mode.
pipetters with one-stroke mechanisms positive displacement
7.3.2 Aspiration—Aspirate in accordance with 7.2.2, except
pipetters to errors introduced by slight variations of the
that the push-button is depressed to the bottom stop position
dynamicsoftheliquidinterfacebreakattheendofthepipetor
prior to pipet tip immersion.
pipettipduringthedispensingaction,duetothepurgingaction
7.3.3 Delivery:
of the air “blow-out” stroke potential.
7.3.3.1 Place the pipet or pipet tip at an angle (10 to 45°, or
7.1.3 The use of the reverse mode with two-component
as prescribed by the manufacturer) against the inside wall of
stroke mechanism pipetters may be more advantageous when
the receiving vessel.
liquids that are difficult to handle in the forward mode are
7.3.3.2 Depress the push-button smoothly to the intermedi-
encountered.
ate stop position.
7.2 Forward Mode, General Format:
7.3.3.3 After a 1-s wait, remove the pipet or pipet tip from
7.2.1 Preparation—Pipetter and environment shall be iso-
the sidewall, in accordance with 7.2.3.
thermal. Volume settings and the mounting of removable or
7.3.3.4 In the case of the pipet tip being reused, allow the
disposable pipet tips shall be accomplished according to the
push-button to remain in the intermediate stop position for
manufacturer’s directions.
subsequent immersion for the next pipetting cycle. In the case
7.2.2 Aspiration:
of the pipet tip to be changed, allow the push-button to return
7.2.2.1 Hold the instrument in a vertical position, or as
to the top stop position.
prescribed by the manufacturer.
NOTE 2—Top and bottom stop positions, as described in the procedures
7.2.2.2 In the case of two-component stroke systems, de-
above,arenotmeanttoincludeauxiliarystrokepositions(forexample,for
press the push button smoothly to the intermediate stop
tip ejection).
position.
7.4 Prerinsing (Forward Mode):
7.2.2.3 In the case of one-component stroke systems, de-
7.4.1 Prerinsing is the action of precoating the inside of the
press the push-button smoothly to the bottom stop position.
liquid contracting part(s) with a thin film of the same liquid to
7.2.2.4 Immerse the pipet or pipet tip into the liquid to be
be pipetted. It is accomplished by duplicating the exact motion
pipetted to, and maintain it at the following depth:
of a forward mode pipetting cycle, except that the liquid is
Volume, µL Immersion Depth, mm
dispensedbackintotheoriginalvessel,orpreferablydiscarded.
1to100 2to3
7.4.2 Prerinsing in the forward mode is advantageous when
101 to 1000 2 to 4
1.1to10mL 3 to 6
reusing (the same liquid and volume setting only) the pipet or
pipet tip for subsequent immediate pipettings. Eliminating the
7.2.2.5 Allow the push-button to move up to the top stop
dispensedamountfromthefirstwettingfromthesamplegroup
position slowly and smoothly.
formed by subsequent wettings and thus the removal of its
7.2.2.6 For air displacement pipetters, observe a wait of 1 s.
valuefromthecalculationofaprecisionstatisticforthegroup,
7.2.2.7 Withdraw the pipet or pipet tip smoothly by lifting
will result in a more precise distribution.
straight up either from the center of the liquid surface in the
7.4.3 Prerinsing may also be practice
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