ASTM E1154-89(2003)
(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 , 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: E 1154 – 89 (Reapproved 2003)
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 (e) indicates an editorial change since the last revision or reapproval.
1. Scope 3.1.3 disposable—those parts of an instrument that are
intended to be used once only and then discarded. Disposable
1.1 This specification covers requirements, operating con-
parts are generally intended for use in applications where
ditions, and test methods for piston or plunger operated
sample carryover is intolerable.
volumetric apparatus (POVA).
3.1.4 maximum error—the maximum difference between
1.2 This specification includes specifications applicable for
thenominalvolumeandanysingleindividualvolumeobtained
all types of POVA or those given by the manufacturer. The
by applying the test procedure specified in Section 13 of this
followingprecautionarycaveatpertainsonlytothetestmethod
ISO Standard.
portion, Section 13, of this specification: This standard does
3.1.5 maximum expectable error—with more than 95%
not purport to address all of the safety concerns, if any,
probability, the maximum expectable error is calculated as
associated with its use. It is the responsibility of the user of this
follows:
standard to establish appropriate safety and health practices
and determine the applicability of regulatory limitations prior
6~1E 1 12s! (1)
T
to use.
where:
2. Referenced Documents
E = inaccuracy of the mean, and
2 T
2.1 ISO Documents:
s = standard deviation from the repeatability test in
ISO3534 Statistics—Vocabulary and Symbols
Section 13.
ISO653 Long Solid-Stem Thermometers for Precision Use
3.1.6 nominal volume(s)—the stated volume(s) for which
ISO655 Long Enclosed-Scale Thermometers for Precision
performance is specified.
Use
3.1.7 unit of volume—themillilitreorthemicrolitre,thatare
ISO4787 Laboratory Glassware—Volumetric Glassware—
accepted substitutes for the cubic centimetre or cubic millime-
Methods for Testing and Use
tre.
3.1.7.1 Discussion—Itisrecommendedthatvolumesshould
3. Terminology
be specified in microlitres up to 999 µL, and in millilitres from
3.1 Definitions of Terms Specific to This Standard:
1 mL.
3.1.1 accuracy —the accuracy of an instrument is the
3.1.8 piston or plunger operated volumetric apparatus
closeness of agreement between the nominal volume and the
(POVA)—the volume of liquid to be measured with POVA is
mean volume, obtained by applying the test procedure speci-
defined by one or more strokes of one or more pistons or
fied in Section 13 of this specification. It is quantified by the
plungers. POVA may be operated manually or mechanically
inaccuracy of the mean.
(for example, electrically, pneumatically or by hydrostatic
3.1.2 dead volume—thedeadvolumeisthatpartofthetotal
pressure).
liquidvolume,heldintheoperationalpartofthedevice,which
3.1.8.1 Discussion—In the following text the word8 piston’
is not delivered.
means8 piston or plunger.’
3.1.2.1 Discussion—The dead volume should not be con-
3.1.9 precision —the closeness of agreement between the
fused with the dead space of an air displacement instrument.
individual volumes obtained by applying the test procedure
specified in this specification. It is quantified by the impreci-
sion.
This specification is under the jurisdiction of ASTM Committee E41 on
3.1.9.1 Discussion—Thetestprocedurespecifiedgivesonly
Laboratory Apparatus and is the direct responsibility of Subcommittee E41.06 on
a measure of the repeatability (see ISO 3534) under controlled
Weighing Devices.
Current edition approved April 4, 1989. Published May 1989. Originally
conditions.
published as E1154-87. Last previous edition E1154-87.
3.1.10 reusable—thosepartsofaninstrumentthataremeant
Available from American National Standards Institute, 1430 Broadway, New
tobeusedmorethanonce.Asthereusabilityofsomepartscan
York, NY 10018.
These definitions apply only in the cases where the distributions are Gaussian. rarely be quantified, any institution or individual who reuses a
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
E 1154 – 89 (2003)
reusable part must see to its safety and effectiveness. Reusable liquid interface (indirect action) or by actual contact with the
parts are generally intended for use in applications where piston (direct action), (see Fig. 3 and Fig. 4).
sample carryover is tolerable, or can be adequately prevented.
5. Performance Requirements
3.1.11 sample carryover—that portion of the sample that is
retained in the instrument and that may affect subsequent
5.1 Performance Tolerances:
samples. 5.1.1 Performance tolerances specified for POVAare meant
3.1.12 stated feature—any feature claimed by the manufac-
to include any thermal drift effect upon the accuracy and
turer. precision attributable to hand-transmitted heat during normal
3.1.13 reference temperature—the temperature at which the
use. It is, therefore, important that the instrument being
instrument is designed to deliver its nominal volume(s). evaluated according to the referenced procedure not be pre-
3.1.13.1 Discussion—At that temperature the closest agree- conditioned (warmed) by recent handling, nor isolated from
ment between manufacturer’s performance claims and test normal handwarming during the test series (30 or 10 cycles).
results may be expected. 5.1.2 Volumetricperformancetolerancesarenotspecifiedin
3.1.14 reference temperature range—that temperature this specification. The manufacturer shall specify the perfor-
¯
range for which the tolerances for accuracy are specified. mancetolerancesintermsoftheaccuracyofthemean(E %)
C
3.1.15 working range—that part (of the total range) for
andcoefficientofvariation(CV %).Valuesshallbegivenfor
c
which manufacturer’s performance specifications are given. 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 5.2 The reference temperature recommended for all POVA
given.
is 21.5°C, which is the mid-point of the reference temperature
range, (see section 3.1.14). The use of another reference
4. Classification
temperature must be stated by the manufacturer.
4.1 Types of POVA—Piston or plunger operated volumetric 5.2.1 Reference Temperature Range— The reference tem-
apparatus (POVA) are classified as follows: perature range for all POVA shall be 19 to 24°C, (see section
4.1.1 Pipette—Ameasuring instrument for the transfer of a 3.1.13 and section 3.1.14).
predetermined volume of liquid from one vessel to another. It 5.3 Removable Parts:
is not connected to a reservoir. 5.3.1 The volumetric performance of POVAto be used with
4.1.2 Dispenser—A measuring instrument for delivering removable parts can depend to a large extent on the design,
predetermined volumes of liquid from a reservoir. The reser- material, and workmanship of those parts. The test procedures
voir may be integrated with the instrument or connected described can give information only about the performance of
externally. the instruments together with the removable parts actually
4.1.3 Dilutor—Ameasuring instrument for taking up differ- used.
ent liquids (for example, sample and diluent) and delivering 5.3.2 Single-Measurement Test—The single-measurement
them in combination so as to comprise a predetermined ratio, menttestrequireseither30or10randomlyselectedremovable
or predetermined volumes, or both. The reservoir of diluent parts, one for each sample of the series. This test evaluates the
may be integrated with the instrument or connected externally. instrument’s performance and component of imprecision due
4.1.4 Displacement Buret—A measuring instrument from to the variation of these parts.
which the volume delivered is determined by an external 5.3.3 Replicate-Delivery Test—The replicate delivery test
indicator. The volume delivered can then be read. uses one removable part for the 30 or 10 sample series. This
4.2 Types of Displacement: testevaluatestheinstrument’sperformanceandthecomponent
4.2.1 Displacement with an air interface (“air displace- of imprecision due to the reuse of this part.
ment”). The delivered liquid is displaced by an air interface 5.4 Durability—Any claim by a manufacturer that an in-
(indirect action), (see Figs. 1 and 2). strument is resistant to any defined conditions (for example,
4.2.2 Displacement without an air interface (“positive dis- sterilization and chemical exposure) shall be understood in
placement”). The delivered liquid is displaced either by a such a way that even long term or repeated exposure to those
FIG. 1 Displacement With an Air Interface (Air Displacement)
E 1154 – 89 (2003)
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)
conditions(asspecifiedbythemanufacturer)willnotaffectthe 7. Operating Conditions for Pipetters
rated performance of the instrument.
7.1 Twocommonmodesofoperationareinuse,theforward
mode (sometimes referred to as normal mode), and the reverse
6. General Operating Conditions
mode (usable with two-component stroke mechanism systems
only), (see Fig. 3 and Fig. 4).
6.1 Relationship to Performance—The specification of op-
erating procedures is critical to the proper functioning of the
7.1.1 In general, the precision of the repetitive use of the
instruments, and determines their ability to perform within
forward mode relies upon the precise draining by air pressure
specified tolerances. Changes in the operating mode can (in the case of air displacement pipetters) or internal wiping of
dramatically alter the results of analyses. Most instruments are
thepipetbarrelortip(inthecaseofdisplacementpipetters).As
calibrated for certain operating modes; another manner of use compared to the reverse mode, the forward mode is relatively
may result in a change in the accuracy or precision, or both.
insensitivetovariationsinthespeedofthepistonorplungerin
the dispensing action. Positive displacement instruments with
6.2 Delineation—It is the manufacturer’s responsibility to
delineate the modes of operation in instruction manuals and to relatively small delivery orifices are generally less sensitive to
change in accuracy when handling liquids with high wetability
state for which of the modes the instrument is calibrated.
characteristics.
6.3 Preparation—The manufacturer shall provide instruc-
tions necessary for the preparation of the instrument for use in 7.1.2 Airdisplacementpipetterswithtwo-componentstroke
particular operating modes (for example, mounting of remov- mechanisms are generally less sensitive than air displacement
able parts, method of volume adjustment, temperature equa- pipetters with one-stroke mechanisms positive displacement
tion, isothermal requirements, testing of piston action, lubrica- pipetters to errors introduced by slight variations of the
tion, priming, purging or prerinsing information, etc.). dynamicsoftheliquidinterfacebreakattheendofthepipetor
E 1154 – 89 (2003)
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—Topandbottomstoppositions,asdescribedintheprocedures
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-
press the push-button smoothly to the bottom stop position. 7.4.1 Prerinsing is the action of precoating the inside of the
7.2.2.4 Immerse the pipet or pipet tip into the liquid to be liquid contracting part(s) with a thin film of the same liquid to
pipetted to, and maintain it at the following depth: be pipetted. It is accomplished by duplicating the exact motion
of a forward mode pipetting cycle, except that the liquid is
Volume, µL Immersion Depth, mm
1to100 2to3
dispensedbackintotheoriginalvessel,orpreferablydiscarded.
101 to 1000 2 to 4
7.4.2 Prerinsing in the forward mode is advantageous when
1.1to10mL 3 to 6
reusing (the same liquid and volume setting only) the pipet or
7.2.2.5 Allow the push-button to move up to the top stop
pipet tip for subsequent immediate pipettings. Eliminating the
position slowly and smoothly.
dispensedamountfromthefirstwettingfromthesamplegroup
7.2.2.6 For air displacement pipetters, observe a wait of 1 s.
formed by subsequent wettings and thus the removal of its
7.2.2.7 Withdraw the pipet or pipet tip smoothly by lifting
valuefromthecalculationofaprecisionstatisticforthegroup,
straight up either from the center of the liquid surface in the
will result in a more precise distribution.
vessel, or up the sidewall of the vessel.
7.4.3 Prerinsing may also be practiced when a removable
pipet tip is to be used only once (for example, when pipetting
NOTE 1—No further liquid contact of the pipet or pipet tip is allowed
once the liquid interface is broken.
different liquids), but the increase in time required to accom-
modate prerinsing each tip reserves this practice for pipetting
7.2.2.8 Wipe the pipet or pipet tip only if t
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