近日，EDQM-OMCL發(fā)布了《活塞式容量?jī)x器（POVA）確認(rèn)指南》，該指南描述了用于化學(xué)和生物測(cè)試的 POVA（如移液器）的要求，包含活塞式容量?jī)x器（如移液槍?zhuān)┦褂玫淖罴褜?shí)踐以及人員確認(rèn)要求、活塞式容量?jī)x器（如移液槍?zhuān)筛鼡Q部件）的選擇考慮、使用前考慮、校準(zhǔn)要求、校準(zhǔn)前的初步檢查、計(jì)量確認(rèn)、環(huán)境條件、日常測(cè)試的一般考慮、活塞式容量?jī)x器（POVA）的評(píng)估、合格 / 不合格狀態(tài)的判定、要求及相關(guān)典型接受標(biāo)準(zhǔn)等。

解析如下：

指南將移液槍的測(cè)試分為校準(zhǔn)和日常測(cè)試：

校準(zhǔn)：建立交付容量與儀器選定容量之間關(guān)系以及相關(guān)測(cè)量不確定度的一系列操作。校準(zhǔn)不需要對(duì)儀器進(jìn)行永久性修改的操作，也不需要對(duì)設(shè)備進(jìn)行調(diào)整。

日常測(cè)試：定期（例如每三個(gè)月）進(jìn)行的分析設(shè)備監(jiān)測(cè)或分析質(zhì)量控制程序。該術(shù)語(yǔ)與 “確認(rèn)（verification）” 同義。

測(cè)量不確定度：進(jìn)行校準(zhǔn)時(shí)，應(yīng)評(píng)估并報(bào)告測(cè)量不確定度；而在日常測(cè)試中，則是可選的。

使用移液器時(shí)，使用人員的移液技術(shù)通常是容量誤差的最大來(lái)源。因此，使用、測(cè)試或校準(zhǔn)活塞式容量?jī)x器（POVA）的合適人員應(yīng)接受液體處理工具選擇和正確移液技術(shù)方面的培訓(xùn)。實(shí)驗(yàn)室應(yīng)建立并記錄活塞式容量?jī)x器（POVA，如移液槍?zhuān)┦褂萌藛T的能力要求，包括確認(rèn)和培訓(xùn)要求。使用人員能力可通過(guò)日常測(cè)試、能力驗(yàn)證或使用人員間比對(duì)等方式進(jìn)行監(jiān)測(cè)。

移液槍的校準(zhǔn)前檢查：

活塞功能：平穩(wěn)且可靠。

頭座：無(wú)痕跡或變形，無(wú)液體殘留。

泄漏：吸取標(biāo)稱(chēng)容量 10 秒后，吸頭處無(wú)液滴形成。

通過(guò)目視檢查，無(wú)內(nèi)部或外部污染。

應(yīng)在以下情況對(duì)活塞式容量?jī)x器（POVA，如移液槍?zhuān)┻M(jìn)行校準(zhǔn)：

收到時(shí)（除非供應(yīng)商已校準(zhǔn)）；

在移液器使用期間定期進(jìn)行，且至少每 12 個(gè)月一次；

在任何維護(hù)、修理或調(diào)整之后。

實(shí)驗(yàn)室可根據(jù)風(fēng)險(xiǎn)評(píng)估，考慮進(jìn)行更頻繁和定期的校準(zhǔn)

校準(zhǔn)資質(zhì)要求：如校準(zhǔn)由服務(wù)提供商進(jìn)行，實(shí)驗(yàn)室應(yīng)確認(rèn)并選擇一家依據(jù) ISO/IEC 17025 獲得認(rèn)可的提供商，以按照 ISO 8655 的要求進(jìn)行校準(zhǔn)；如果校準(zhǔn)由實(shí)驗(yàn)室內(nèi)部操作人員進(jìn)行，應(yīng)選擇合適的校準(zhǔn)程序。

文件給出校準(zhǔn)程序的要求：

校準(zhǔn)應(yīng)反映日常操作（例如，使用的操作范圍；用于移液器校準(zhǔn)的吸頭最好與日常工作中使用的類(lèi)型相同 - 對(duì)于可滅菌吸頭，滅菌程序不應(yīng)影響吸頭的計(jì)量特性。

例外情況是移液模式，校準(zhǔn)應(yīng)僅以正向模式進(jìn)行；反向模式校準(zhǔn)不再被接受。

對(duì)于日常測(cè)試，反向模式是可接受的。

校準(zhǔn)必須在受控的環(huán)境條件（即溫度、濕度和壓力）下進(jìn)行，以盡量減少水的蒸發(fā)，尤其是容量低于 50 μL 的移液器。

環(huán)境條件要求：

在開(kāi)始測(cè)試前至少 2 小時(shí)（最短平衡時(shí)間）以及測(cè)試期間，測(cè)試室的空氣溫度和相對(duì)濕度應(yīng)在規(guī)定的范圍內(nèi)。

 

環(huán)境條件	ISO 8655 - 6、8*	ISO 8655 - 7**
測(cè)試室溫度	20 ± 3℃	需記錄
測(cè)試期間溫度變化	0.5℃	0.5℃，需記錄
2 小時(shí)平衡期間溫度變化	0.5℃/ 小時(shí)	1℃，需記錄
水 / 測(cè)試液體溫度	環(huán)境空氣溫度 ± 0.5℃	需記錄
測(cè)試室濕度	45 - 80%	需記錄
大氣壓力	需記錄	需記錄

 

應(yīng)避免長(zhǎng)時(shí)間（超過(guò) 10 分鐘）使用移液器導(dǎo)致其被手捂熱。來(lái)自手的熱傳遞可能導(dǎo)致熱不平衡，并影響移液器氣墊的容量。

文件給出各種類(lèi)型移液器的最大允許誤差：

A型和D1型（單通道移液器）的最大允許誤差（MPE）

A型和DI型（多通道移液器）的最大允許誤差

D2型的最大允許誤差

單步移液器的最大允許誤差

 

多步遞送分配器的最大允許誤差

 

翻譯如下：

QUALIFICATION OF PISTON - OPERATED VOLUMETRIC APPARATUS (POVA)活塞式容量?jī)x器（POVA）的確認(rèn)
Contents目錄Introduction引言Aim and Scope of the Guideline指南的目的和范圍Glossary (ISO 8655 - 1)術(shù)語(yǔ)（ISO 8655-1）Best practices for the use of POVA and requirements for staff qualification活塞式容量?jī)x器（POVA，如移液槍?zhuān)┦褂玫淖罴褜?shí)踐以及人員確認(rèn)要求General Consideration for LEVEL I on selection of POVA (including exchangeable parts)LEVEL I ：活塞式容量?jī)x器（如移液槍?zhuān)筛鼡Q部件）選擇的一般考慮General Consideration for LEVEL III - Release for useLEVEL III：投入使用的一般考慮General Consideration for LEVEL II - CalibrationLEVEL II：校準(zhǔn)的一般考慮7.1 Preliminary checks before calibration7.1 校準(zhǔn)前的初步檢查7.2 Metrological confirmation7.2 計(jì)量確認(rèn)7.3 Environmental conditions7.3 環(huán)境條件7.4 Gravimetric test7.4 重量測(cè)試General Consideration for LEVEL IV - Routine testsLEVEL IV - 日常測(cè)試的一般考慮8.1 Preliminary checks before routine testing8.1 日常測(cè)試前的初步檢查8.2 Metrological confirmation8.2 計(jì)量確認(rèn)POVA Evaluation活塞式容量?jī)x器（POVA）的評(píng)估9.1 Determination of pass/fail status9.1 合格 / 不合格狀態(tài)的判定9.2 Non - conformity of pipettes9.2 移液器的不符合項(xiàng)9.3 Calibration/report9.3 校準(zhǔn) / 報(bào)告9.4 Requirements and related typical acceptance criteria9.4 要求及相關(guān)典型接受標(biāo)準(zhǔn)
Introduction引言
The present document is the 6th Annex of the core document "Qualification of Equipment" [1], and it should be used in combination with it when planning, performing and documenting the qualification of piston - operated volumetric apparatus (POVA).本文件是核心文件 “設(shè)備確認(rèn)”[1] 的第 6 個(gè)附錄，在規(guī)劃、實(shí)施和記錄活塞式容量?jī)x器（POVA）的確認(rèn)時(shí)，應(yīng)與該核心文件結(jié)合使用。
The core document "Qualification of Equipment" contains the Introduction and general forms for Level I and II of qualification, which are common to all types of instruments.核心文件 “設(shè)備確認(rèn)” 包含引言以及LEVEL I 和LEVEL II 確認(rèn)的通用表格，他們對(duì)所有類(lèi)型的儀器都是通用的。
Level III and IV testing must be carried out, as an ISO/IEC 17025 [2] requirement, by trained personnel.根據(jù) ISO/IEC 17025 [2] 的要求，LEVEL III 和LEVEL IV測(cè)試必須由經(jīng)過(guò)培訓(xùn)的人員進(jìn)行。
This Guideline has been elaborated according to the standard ISO 8655 (Parts - 1, - 2, - 5, - 6, - 7, - 8 and - 10) [3 - 9]. Requirements and (if applicable) corresponding acceptance criteria should be applied; however, other appropriately justified deviations are acceptable provided they are traceable.本指南是根據(jù) ISO 8655 標(biāo)準(zhǔn)（第 1、2、5、6、7、8 和 10 部分）[3 - 9] 制定的。應(yīng)應(yīng)用相關(guān)要求以及（如適用）相應(yīng)的接受標(biāo)準(zhǔn)；然而，可以接受其他經(jīng)論證的偏差，應(yīng)可以追溯。
This document describes the method given in ISO 8655-6 "Gravimetric reference measurement procedure for the determination of volume" [6], which is the most commonly used.本文件描述了 ISO 8655 - 6《用于容量測(cè)定的重量法參考測(cè)量程序》[6] 中給出的方法，這是最常用的方法。
Another reference measurement procedure is described in ISO 8655-8 "Photometric reference measurement procedure for the determination of volume" [8], which is applicable to apparatus with a maximum nominal volume of 5 000 μL.另一種參考測(cè)量程序在 ISO 8655 - 8《用于容量測(cè)定的光度法參考測(cè)量程序》[8] 中描述，其適用于最大標(biāo)稱(chēng)容量為 5000 微升的儀器。
Alternative procedures are given in ISO 8655-7 "Alternative measurement procedures for the determination of volume" [7].替代程序在 ISO 8655 - 7《用于容量測(cè)定的替代測(cè)量程序》[7] 中給出。
In this Annex the general requirements for metrological confirmation of POVAs, which comprises calibration and verification, are described, as well as other requirements included in ISO 8655-10 "User guidance, and requirements for competence, training, and POVAsuitability" [9].在本附錄中，描述了 POVA（移液器）計(jì)量確認(rèn)的一般要求，包括校準(zhǔn)和驗(yàn)證，以及 ISO 8655 - 10《使用人員指南以及能力、培訓(xùn)和 POVA 適用性要求》[9] 中包含的其他要求。
Aim and Scope of the Guideline指南的目的和范圍
This guideline describes the requirements for POVAs used in chemical and biological tests irrespective of the mode of operation (manual or electronic).本指南描述了用于化學(xué)和生物測(cè)試的 POVA（移液器）的要求，無(wú)論其操作模式（手動(dòng)或電動(dòng)）如何。
Requirements and test procedures are based on the ISO 8655 standard.要求和測(cè)試程序基于 ISO 8655 標(biāo)準(zhǔn)。
The following types of pipettes have been considered in this guideline:本指南考慮了以下類(lèi)型的移液器：
 1. Fixed - volume single - channel pipettes (air displacement (type A) and positive displacement or direct displacement pipettes (type D, with a reusable or disposable plunger and capillary)) 1. 固定容量單通道移液器（空氣置換式（A 型）和正置換式或直接置換式移液器（D 型，帶有可重復(fù)使用或一次性活塞和毛細(xì)管））
 2. Variable - volume single - channel pipettes (air displacement (type A) and positive displacement or direct displacement pipettes (type D1) reusable or (type D2) disposable plunger and capillary) 2. 可變?nèi)萘繂瓮ǖ酪埔浩鳎諝庵脫Q式（A 型）和正置換式或直接置換式移液器（D1 型可重復(fù)使用或 D2 型一次性活塞和毛細(xì)管））
 3. Fixed - volume multi - channel pipettes (air displacement (type A)) 3. 固定容量多通道移液器（空氣置換式（A 型））
 4. Variable - volume multi - channel pipettes (air displacement (type A)) 4. 可變?nèi)萘慷嗤ǖ酪埔浩鳎諝庵脫Q式（A 型））
 5. Electronic motorized pipettes 5. 電動(dòng)移液器
 6. Dispensers (e.g. repetitive piston pipettes) 6. 分配器（例如重復(fù)活塞移液器）
NOTES:注釋?zhuān)?br /> Although designed as variable - volume pipettes, electronic motorized pipettes may be used in different modes:盡管電動(dòng)移液器設(shè)計(jì)為可變?nèi)萘恳埔浩鳎稍诓煌Ｊ较率褂茫?/span>

Pipetting mode: the selected volume is aspirated and dispensed in forward mode pipetting.

移液模式：以正向移液模式吸取和分配所選容量。

Multi - dispensing mode: the aspirated volume is dispensed repeatedly in a predefined number of aliquots, which can be of equal or different volumes.

多次分配模式：吸取的容量以預(yù)定義數(shù)量的等分試樣重復(fù)分配，這些等分試樣的容量可以相等或不同。

Mixing mode: pipetting step is followed by a mixing phase, composed of repeated aspirating and dispensing.

混合模式：移液步驟之后是混合階段，由重復(fù)的吸取和分配組成。

Reverse mode: aspiration of the selected volume added with an extra amount of liquid that remains in the tip after the dispensing of the selected volume. The remaining volume can subsequently be discarded.

反向模式：吸取所選容量并額外添加一定量的液體，該液體在分配所選容量后殘留在吸頭中。剩余容量隨后可以丟棄。

Multi - aspirated: aspiration of a predefined number of equal or different volumes, followed by the dispense of the total aspirated volume.

多次吸取：吸取預(yù)定義數(shù)量的相等或不同容量，然后分配總共吸取的容量。

Additionally, this guideline provides guidance, based on ISO 8655-10 [9] regarding:
此外，本指南基于 ISO 8655 - 10 [9] 提供以下方面的指導(dǎo)：

The selection of POVAs (including best practices for their use);

活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;的選擇（包括其使用的最佳實(shí)踐）；

User qualification/re - qualification (requirements for user training and competence);

使用人員確認(rèn) / 重新確認(rèn)（使用人員培訓(xùn)和能力要求）；

POVA performance requirements and tolerances to ensure fitness for their intended use;

POVA（移液器）性能要求和公差，以確保符合其預(yù)期用途；

Replicate measurements and uncertainty.

重復(fù)測(cè)量和不確定度。

3. Glossary (ISO 8655 - 1)3. 術(shù)語(yǔ)（ISO 8655 - 1）
Calibration: set of operations that establish the relationship between the delivered volume and the corresponding selected volume of the apparatus with associated measurement uncertainties.校準(zhǔn)：建立交付容量與儀器選定容量之間關(guān)系以及相關(guān)測(cè)量不確定度的一系列操作。
NOTE:注釋?zhuān)?br /> Calibration requires no operation which permanently modifies the apparatus and does not require adjustment of the device.校準(zhǔn)不需要對(duì)儀器進(jìn)行永久性修改的操作，也不需要對(duì)設(shè)備進(jìn)行調(diào)整。
Routine testing: testing equipment monitoring or analytical quality - control routines to be performed at regular intervals, for example, every three months. Other time intervals may be adopted giving due consideration to the factors described in Level IV (paragraph 8). This term is a synonym of "verification".日常測(cè)試：定期（例如每三個(gè)月）進(jìn)行的分析設(shè)備監(jiān)測(cè)或分析質(zhì)量控制程序。在適當(dāng)考慮 IV 級(jí)（第 8 段）所述因素的情況下，可采用其他時(shí)間間隔。該術(shù)語(yǔ)與 “確認(rèn)（verification）” 同義。
Measurement uncertainty: non - negative parameter, associated with the delivered volume, that characterises the dispersion of the volumes that could reasonably be attributed to the delivered volume based on the information used.測(cè)量不確定度：與交付容量相關(guān)的非負(fù)參數(shù)，用于表征基于所用信息合理歸因于交付容量的容量分散性。
When performing calibration, the measurement uncertainty shall be evaluated and reported; however, in routine testing it is optional.進(jìn)行校準(zhǔn)時(shí)，應(yīng)評(píng)估并報(bào)告測(cè)量不確定度；而在日常測(cè)試中，則是可選的。
Metrological confirmation: set of operations required to ensure that the POVA conforms to the requirements for its intended use.計(jì)量確認(rèn)：為確保活塞式容量?jī)x器（POVA）符合其預(yù)期用途要求所需的一系列操作。
NOTE:注釋?zhuān)?br /> Metrological confirmation generally includes calibration and verification, and any necessary adjustment or repair, and subsequent recalibration, comparison with the metrological requirements for the intended use of the POV A, as well as any required sealing and labelling.計(jì)量確認(rèn)通常包括校準(zhǔn)和驗(yàn)證，以及任何必要的調(diào)整或維修，隨后的再校準(zhǔn)，與 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;預(yù)期用途的計(jì)量要求進(jìn)行比較，以及任何必要的密封和貼標(biāo)簽。
Nominal volume: upper limit of the usable volume range as specified by the manufacturer.標(biāo)稱(chēng)容量：制造商規(guī)定的可用容量范圍的上限。
NOTES:注釋?zhuān)?br /> For a variable - volume piston - operated volumetric apparatus, the nominal volume corresponds to the maximum volume that can be set by the user and that is specified by the manufacturer.對(duì)于可變?nèi)萘炕钊饺萘績(jī)x器，標(biāo)稱(chēng)容量對(duì)應(yīng)于使用人員可設(shè)置且由制造商規(guī)定的最大容量。
For dispensers the nominal volume is depending on the tips used.對(duì)于分配器，標(biāo)稱(chēng)容量取決于所用的吸頭。
For the other terms refer to JCGM 200:2012 International vocabulary of metrology - Basic general concepts and associated terms (VIM) [10] and ISO 8655 - 1 [3].其他術(shù)語(yǔ)請(qǐng)參考 JCGM 200:2012《國(guó)際計(jì)量學(xué)詞匯 - 基本通用概念及相關(guān)術(shù)語(yǔ)》（VIM）[10] 和 ISO 8655 - 1 [3]。
4. Best practices for the use of POVA and requirements for staff qualification活塞式容量?jī)x器（POVA）的最佳使用實(shí)踐和人員確認(rèn)要求
Guidance for the use of the different POV A, pipetting technique, as well as references to best practices for handling non - aqueous liquids are given in ISO 8655 - 10 [9]. Critical aspects to take into account, if applicable, include:ISO 8655 - 10 [9] 給出了不同活塞式容量?jī)x器（POVA）的使用指南、移液技術(shù)，以及處理非水液體的最佳實(shí)踐參考。如適用，需要考慮的關(guān)鍵方面包括：

Setting of the volume;

容量設(shè)置；

Pre - wetting of pipette tips/Immersion depth;

移液器吸頭的預(yù)濕潤(rùn) / 浸入深度；

Forward and reverse pipetting modes;

正向和反向移液模式；

Thermal equilibrium/Hand warming;

熱平衡 / 手溫預(yù)熱；

Tip position during aspiration and dispense/Pause after aspiration;

吸取和分配過(guò)程中的吸頭位置 / 吸取后暫停；

Velocity of aspiration and delivery;

吸取和排出速度；

Tip wiping/Cleaning/Priming.

吸頭擦拭 / 清潔 / 預(yù)充。

The ability to consistently deliver precise and accurate volumes is the volumetric performance, it is directly dependent on the system, the POV A and the technique employed by the user of the pipette.始終如一地提供精確準(zhǔn)確容量的能力是容量性能，它直接取決于系統(tǒng)、活塞式容量?jī)x器（POVA）以及移液器使用人員采用的技術(shù)。
When using pipettes, the user's pipetting technique is usually the largest contributor to volumetric error. Therefore, proper users who use, test or calibrate POV A shall be trained on liquid handling tool selection and proper pipetting techniques.使用移液器時(shí)，使用人員的移液技術(shù)通常是容量誤差的最大來(lái)源。因此，使用、測(cè)試或校準(zhǔn)活塞式容量?jī)x器（POVA）的合適人員應(yīng)接受液體處理工具選擇和正確移液技術(shù)方面的培訓(xùn)。
The laboratory shall establish and document the competence requirements for POV A users, including qualification and training requirements.實(shí)驗(yàn)室應(yīng)建立并記錄 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;使用人員的能力要求，包括確認(rèn)和培訓(xùn)要求。
User competence can be monitored, e.g. by routine testing, proficiency testing or as inter - comparison between users.使用人員能力可通過(guò)日常測(cè)試、能力驗(yàn)證或使用人員間比對(duì)等方式進(jìn)行監(jiān)測(cè)。
A report of training or competence assessment can be issued to the user, whose content should include:可向使用人員出具培訓(xùn)或能力評(píng)估報(bào)告，其內(nèi)容應(yīng)包括：

Identification of the user

使用人員身份信息

Indication whether it is a training (trainer’s identification) or competence assessment report

表明是培訓(xùn)報(bào)告（培訓(xùn)人員身份識(shí)別）還是能力評(píng)估報(bào)告

Identification of the POV A and tips used

所用 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;及吸頭的編號(hào)

Environment of test conditions (at least room temperature and air relative humidity – RH)

測(cè)試條件的環(huán)境（至少包括室溫及空氣相對(duì)濕度 - RH）

Volumes tested

測(cè)試的容量

Criteria of the operator competence assessment (e.g. systematic and random errors or uncertainties in use of a single delivered volume for the tested volumes)

操作人員能力評(píng)估標(biāo)準(zhǔn)（例如，針對(duì)測(cè)試容量，使用單次交付容量時(shí)的系統(tǒng)誤差和隨機(jī)誤差或不確定度）

Reference to ISO 8655 - 10 [9] (and any deviations) as well as the employed test method (e.g. ISO 8655 - 6 [6], ISO 8655 - 8 [8] or any specific test procedure in ISO 8655 - 7 [7])

引用 ISO 8655 - 10 [9]（及任何偏差）以及所采用的測(cè)試方法（例如 ISO 8655 - 6 [6]、ISO 8655 - 8 [8] 或 ISO 8655 - 7 [7] 中的任何特定測(cè)試程序）

Volumetric measurement results for each delivered volume

每個(gè)交付容量的容量測(cè)量結(jié)果

Total number of replicate measurements made for each tested volume, and the number of measurement results used for statistical analysis

每個(gè)測(cè)試容量的重復(fù)測(cè)量總數(shù)，以及用于統(tǒng)計(jì)分析的測(cè)量結(jié)果數(shù)量

Date of test and duration of validation

測(cè)試日期和驗(yàn)證持續(xù)時(shí)間

5. General Consideration for LEVEL I on selection of POV A (including exchangeable parts)5. LEVEL I  活塞式容量?jī)x器（POVA，如移液槍?zhuān)ò筛鼡Q部件）選擇的一般考慮因素
All POV A shall be selected based on their suitability for the intended use. To achieve the best volumetric performance, it is recommended to select a POV A with a nominal volume close to the volume to be delivered. The following factors should be considered when selecting apparatus:所有 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;應(yīng)根據(jù)其對(duì)預(yù)期用途的適用性進(jìn)行選擇。為實(shí)現(xiàn)最佳的容量性能，建議選擇標(biāo)稱(chēng)容量接近待交付容量的 活塞式容量?jī)x器（POVA，如移液槍?zhuān)＿x擇設(shè)備時(shí)應(yīng)考慮以下因素：

smallest and largest liquid volume to be delivered;

待交付的最小和最大液體容量；

liquid properties;

液體特性；

the resulting impact of delivering inaccurate volumes;

交付不準(zhǔn)確容量所產(chǎn)生的影響；performance requirements (maximum permissible errors – MPEs – and/or process tolerances);

性能要求（最大允許誤差 - MPEs - 和 / 或過(guò)程公差）；

type and size of POV A and its application;

活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;的類(lèi)型和尺寸及其應(yīng)用；

single - channel or multi - channel POV A;

單通道還是多通道 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?/span>

delivery of constant volumes during repeated steps;

重復(fù)步驟中恒定容量的交付；

frequency of use.

使用頻率。

NOTE: Refer to ISO 8655 - 10, Table A.1, for the selection of pipettes.注：移液器的選擇請(qǐng)參考 ISO 8655 - 10 的表 A.1。
Exchangeable parts, such as pipette tips, shall be designed to match the design of the POV A. Changes in material, size of tip orifice, taper (angle), dead air volume and retained liquid impacts the performance of the pipetting system. The overall system performance (POVA and exchangeable parts) shall be suitable for its intended purpose.可更換部件，如移液器吸頭，應(yīng)設(shè)計(jì)成與 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;的設(shè)計(jì)相匹配。材料、吸頭孔尺寸、錐度（角度）、死腔容量和殘留液體的變化會(huì)影響移液系統(tǒng)的性能。整個(gè)系統(tǒng)性能（POVA 和可更換部件）應(yīng)適合其預(yù)期用途。
NOTE: Refer to ISO 8655 - 10, Table A.2, for the selection of pipette tips.注：移液器吸頭的選擇請(qǐng)參考 ISO 8655 - 10 的表 A.2。
6. General Consideration for LEVEL II - Release for use6. LEVEL II  - 放行使用的一般考慮
Check if all requirements set up during the selection are met and all necessary requirements are covered and fulfilled in the provided certificate.檢查在選擇過(guò)程中設(shè)定的所有要求是否得到滿足，以及所提供的證書(shū)中是否涵蓋并滿足所有必要要求。
7. General Consideration for LEVEL III - Calibration7. LEVEL III 校準(zhǔn)的一般考慮
 7.1 Preliminary checks before calibration 7.1 校準(zhǔn)前的初步檢查
Correct functionality of POV A shall be checked before calibration, as follows:校準(zhǔn)前應(yīng)檢查 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;的功能是否正常，如下所示：

 Function of piston: smooth and positive.

 活塞功能：平穩(wěn)且可靠。

Tip holder: no marks or distortions, no liquid residues.

頭座：無(wú)痕跡或變形，無(wú)液體殘留。

 Leakage: no drop is formed at the tip after 10 seconds from withdrawing the nominal volume.

 泄漏：吸取標(biāo)稱(chēng)容量 10 秒后，吸頭處無(wú)液滴形成。

No internal or external contamination by visual checking.

 通過(guò)目視檢查，無(wú)內(nèi)部或外部污染。

7.2 Metrological confirmation7.2 計(jì)量確認(rèn)
Some piston pipettes require the use of exchangeable parts during typical use (e.g. disposable or reusable pipette tips), that are considered to be an integral part of the POV A under test.一些活塞式移液器在正常使用過(guò)程中需要使用可更換部件（例如一次性或可重復(fù)使用的移液器吸頭），這些部件被視為被測(cè) 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;的一個(gè)組成部分。
The periodic calibration of POV A shall follow a pre - established calibration plan, within the framework of the Quality Management System of the laboratory.活塞式容量?jī)x器（POVA，如移液槍?zhuān)┑亩ㄆ谛?zhǔn)應(yīng)在實(shí)驗(yàn)室質(zhì)量管理體系的框架內(nèi)，遵循預(yù)先制定的校準(zhǔn)計(jì)劃。
Calibration of the POV A shall be performed:應(yīng)在以下情況對(duì)活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;進(jìn)行校準(zhǔn)：

on receipt (unless already calibrated by the supplier);

收到時(shí)（除非供應(yīng)商已校準(zhǔn)）；

on a regular basis and at least once every 12 months during the use of the pipette;

在移液器使用期間定期進(jìn)行，且至少每 12 個(gè)月一次；

after any maintenance, repair or adjustment.

在任何維護(hù)、修理或調(diào)整之后。

As described in ISO 8655 - 1 [3], the laboratory can consider performing more frequent and regular calibrations based on a risk assessment, depending on:如 ISO 8655 - 1 [3] 所述，實(shí)驗(yàn)室可根據(jù)風(fēng)險(xiǎn)評(píng)估，考慮進(jìn)行更頻繁和定期的校準(zhǔn)，具體取決于：

the workload of the POV A and number of operators using the pipette;

活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;的工作量以及使用移液器的操作人員數(shù)量；

the type of liquids used;

所用液體的類(lèi)型；

 the MPEs selected, depending on the type of use and the accuracy required.

根據(jù)使用類(lèi)型和所需精度選擇的最大允許誤差（MPEs）。

If calibration is carried out by a service provider, the laboratory shall verify and choose a provider that is accredited in accordance with ISO/IEC 17025 [2] to perform calibrations in accordance with the requirements of ISO 8655.如果校準(zhǔn)由服務(wù)提供商進(jìn)行，實(shí)驗(yàn)室應(yīng)確認(rèn)并選擇一家依據(jù) ISO/IEC 17025 [2] 獲得認(rèn)可的提供商，以按照 ISO 8655 的要求進(jìn)行校準(zhǔn)。
If the calibration is performed by internal laboratory operators, the appropriate procedure for calibration shall be selected. Using the gravimetric (ISO 8655 - 6 [6]) or photometric (ISO 8655 - 8 [8]) measurement procedures, the laboratory shall be equipped with suitable balances, in relation to the volume of POV A to be calibrated, or spectrophotometers (plus density and pH meters).如果校準(zhǔn)由實(shí)驗(yàn)室內(nèi)部操作人員進(jìn)行，應(yīng)選擇合適的校準(zhǔn)程序。使用重量法（ISO 8655 - 6 [6]）或光度法（ISO 8655 - 8 [8]）測(cè)量程序時(shí)，實(shí)驗(yàn)室應(yīng)配備與待校準(zhǔn) 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;容量相適應(yīng)的合適天平，或配備分光光度計(jì)（以及密度計(jì)和 pH 計(jì)）。
If the above cannot be fulfilled, the laboratory should consider sub - contracting calibration to an accredited service provider or proceed with alternative measurement procedures for determination of volume as described in ISO 8655 - 7 [7].如果無(wú)法滿足上述要求，實(shí)驗(yàn)室應(yīng)考慮將校準(zhǔn)工作分包給一家認(rèn)可的服務(wù)提供商，或采用 ISO 8655 - 7 [7] 中所述的替代容量測(cè)量程序。
ISO 8655 - 7 includes five test methods and the corresponding test procedures, described in Annex A to Annex E [7]. These methods are suitable for various maximum nominal volumes of POV A (see Table 1) and should be appropriately selected by the user, taking into account that they apply to complete systems comprising the basic apparatus and tips.ISO 8655 - 7 包含五種測(cè)試方法以及相應(yīng)的測(cè)試程序，在附錄 A 至附錄 E [7] 中有描述。這些方法適用于各種最大標(biāo)稱(chēng)容量的 活塞式容量?jī)x器（POVA，如移液槍?zhuān)ㄒ?jiàn)表 1），用戶應(yīng)適當(dāng)選擇，同時(shí)要考慮到這些方法適用于包括基本設(shè)備和吸頭的完整系統(tǒng)。
For calibrations, no less than ten replicate measurements per selected volume shall be performed. Measurement procedures and test conditions shall be validated for their suitability by comparison to one of the reference measurement procedures described in ISO 8655 - 6 [6] or ISO 8655 - 8 [8].在校準(zhǔn)時(shí)，對(duì)于每個(gè)選定的容量，應(yīng)進(jìn)行不少于十次的重復(fù)測(cè)量。測(cè)量程序和測(cè)試條件應(yīng)通過(guò)與 ISO 8655 - 6 [6] 或 ISO 8655 - 8 [8] 中所述的參考測(cè)量程序之一進(jìn)行比較，來(lái)驗(yàn)證其適用性。
 

表I - ISO 8655-7[7]中所述方法所測(cè)試容量范圍匯總

替代測(cè)試方法	ISO 8655 - 7 附錄	測(cè)試容量
1. 重量法	A	0.5 微升至 2000 毫升 *
2. 雙色比率光度法	B	0.1 微升至 5000 微升
3. 單色光度法	C	2 微升至 200 微升
4. 多通道 POVA 的混合光度 / 重量法	D	在 96 孔板中為 1 微升至 100 微升；在 384 孔板中為 1 微升至 50 微升
5. 滴定法	E	≥500 微升

* 基于天平最低要求的容量范圍（見(jiàn)表 3）
Test conditions, equipment and procedure impact the uncertainty of measurement.測(cè)試條件、設(shè)備和程序會(huì)影響測(cè)量的不確定度。
In addition, calibration shall be carried out to reflect routine operation (e.g. operational range of use; tips used for the calibration of pipettes should preferably be the same type as those used in daily work - in the case of sterilizable tips, the sterilization procedures shall not affect the metrological characteristics of the tips (ISO 8655 - 2 [4])). The exception is pipetting mode, where calibration shall only be performed in forward mode; calibration in reverse mode is no longer acceptable.此外，校準(zhǔn)應(yīng)反映日常操作（例如，使用的操作范圍；用于移液器校準(zhǔn)的吸頭最好與日常工作中使用的類(lèi)型相同 - 對(duì)于可滅菌吸頭，滅菌程序不應(yīng)影響吸頭的計(jì)量特性（ISO 8655 - 2 [4]））。例外情況是移液模式，校準(zhǔn)應(yīng)僅以正向模式進(jìn)行；反向模式校準(zhǔn)不再被接受。
For routine testing, reverse mode is acceptable.對(duì)于日常測(cè)試，反向模式是可接受的。
If the laboratory is working with infectious materials, decontamination (e.g. disassembling) should be done before calibration, according to the instructions provided by the manufacturer.如果實(shí)驗(yàn)室正在處理傳染性材料，應(yīng)根據(jù)制造商提供的說(shuō)明在校準(zhǔn)前進(jìn)行去污。
7.3 Environmental conditions7.3 環(huán)境條件
Ambient conditions (i.e. air temperature, RH, atmospheric pressure and water temperature) may influence weighing of the dispensed volume and water density (Z - factor); therefore, the calibration results can be impacted.環(huán)境條件（即空氣溫度、相對(duì)濕度、大氣壓力和水溫）可能會(huì)影響所分配容量的稱(chēng)重和水的密度（Z 因子）；因此，校準(zhǔn)結(jié)果可能會(huì)受到影響。
As a result, calibration must be performed under controlled environmental conditions (i.e. temperature, humidity and pressure) to minimize the evaporation of water, which is critical for pipettes with dispensing volumes lower than 50 μL.因此，校準(zhǔn)必須在受控的環(huán)境條件（即溫度、濕度和壓力）下進(jìn)行，以盡量減少水的蒸發(fā)，這對(duì)于分配容量低于 50 μL 的移液器至關(guān)重要。
Apart from the geometry of the weighing vessel (gravimetric method), the test cycle time (time required to complete the weighing of one delivered volume) is important and shall be kept to a minimum.除了稱(chēng)重容器的幾何形狀（重量法）之外，測(cè)試循環(huán)時(shí)間（完成一次交付容量稱(chēng)重所需的時(shí)間）也很重要，應(yīng)盡量縮短。
Environmental condition requirements are given in Table 2.環(huán)境條件要求見(jiàn)表 2。
Air temperature and RH shall be within the specified limits for the test room for at least 2 h before starting the test (minimum equilibration time) and during the test itself.在開(kāi)始測(cè)試前至少 2 小時(shí)（最短平衡時(shí)間）以及測(cè)試期間，測(cè)試室的空氣溫度和相對(duì)濕度應(yīng)在規(guī)定的范圍內(nèi)。
 

表2 - ISO 8655第6部分和第8部分與第7部分所述環(huán)境條件要求匯總

環(huán)境條件	ISO 8655 - 6、8*	ISO 8655 - 7**
測(cè)試室溫度	20 ± 3℃	需記錄
測(cè)試期間溫度變化	0.5℃	0.5℃，需記錄
2 小時(shí)平衡期間溫度變化	0.5℃/ 小時(shí)	1℃，需記錄
水 / 測(cè)試液體溫度	環(huán)境空氣溫度 ± 0.5℃	需記錄
測(cè)試室濕度	45 - 80%	需記錄
大氣壓力	需記錄	需記錄

用于校準(zhǔn) / 日常測(cè)試時(shí)需記錄（測(cè)試室條件應(yīng)反映使用 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;的環(huán)境條件）。
用于 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;驗(yàn)證時(shí)。
當(dāng)在采用 27℃作為標(biāo)準(zhǔn)參考溫度的國(guó)家需要重復(fù)使用 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;時(shí)，應(yīng)使用 ISO 384 [1] 中推薦的替代溫度代替該數(shù)值。
7.4 Gravimetric test7.4 重量測(cè)試
The principle of the gravimetric method is described in ISO 8655-6 [6].重量法的原理在 ISO 8655 - 6 [6] 中有描述。
If the selected volume is lower than 50 μL, the effect of the evaporation of the water used is significant.如果所選容量低于 50 微升，所用的水的蒸發(fā)影響是顯著的。
Therefore, the loss of mass should be taken into account in the calculation.因此，在計(jì)算中應(yīng)考慮質(zhì)量損失。
Settings:設(shè)置：
The laboratory where the calibration is performed shall be draught free, with RH of 45 - 80 %.進(jìn)行校準(zhǔn)的實(shí)驗(yàn)室應(yīng)無(wú)氣流，相對(duì)濕度為 45 - 80%。
If volumes lower than 50 μL are selected, an evaporation trap or an open vessel containing water shall be placed inside the balance chamber, and the test cycle time shall be kept to a minimum.如果選擇低于 50 微升的容量，應(yīng)在天平室內(nèi)放置一個(gè)蒸發(fā)阱或一個(gè)盛水的敞口容器，并且測(cè)試周期時(shí)間應(yīng)保持最短。
In addition, the temperature of the laboratory shall be 20 ± 3 °C and stable within ± 0.5 °C during calibration.此外，在校準(zhǔn)期間，實(shí)驗(yàn)室溫度應(yīng)為 20 ± 3°C，且在 ± 0.5°C 范圍內(nèi)保持穩(wěn)定。
All the material needed for the test (glassware, water, exchangeable parts, etc.) must be equilibrated for at least 2 h in the room where the test is to be performed.測(cè)試所需的所有材料（玻璃器皿、水、可更換部件等）必須在進(jìn)行測(cè)試的房間內(nèi)平衡至少 2 小時(shí)。
If using a variable - volume POV A, select the test volume; this setting shall not be altered during the test cycle of all replicate measurements.如果使用可變?nèi)萘康?nbsp;活塞式容量?jī)x器（POVA，如移液槍?zhuān)x擇測(cè)試容量；在所有重復(fù)測(cè)量的測(cè)試周期內(nèi)，此設(shè)置不得更改。
Equipment:設(shè)備：
An analytical balance with appropriate characteristics shall be used depending on the selected volume of the POV A under test (according to ISO 8655 - 6 [6]), as given in Table 3.應(yīng)根據(jù)被測(cè) 活塞式容量?jī)x器（POVA，如移液槍?zhuān)?nbsp;的所選容量（根據(jù) ISO 8655 - 6 [6]），使用具有適當(dāng)特性的分析天平，如表 3 所示。