Pipette Calibration Using a Balance (2024)

Pipette Calibration Using a Balance (1)

Pipette performance verifications according to ISO 8655 are essential to ensure your liquid handling precision instrument is working accurately.

To maintain consistent accuracy of pipetted volumes, pipettes should be calibrated and tested regularly. Pipette calibration is usually performed by a specialized service provider, who will provide a calibration certificate that includes an assessment of the pipette’s measurement uncertainty. In the intervals between calibrations, routine pipette testing carried out by the user ensures a pipette continues to perform according to its specifications. A simple routine test quickly confirms that the instrument is still fit for its intended use and drastically reduces risks of out-of-tolerance (OOT) results.

How does weighing relate to pipette performance?

One of the most important methods for checking pipette performance is using a balance to weigh the liquid delivered by the pipette. The volume of the liquid is then calculated and compared to the expected value. The International Organization of Standardization (ISO) details the requirements for the gravimetric reference measurement procedure for the determination of volume. ISO 8655-6 includes minimum requirements for balances according to the nominal volume of the pipette under test. ISO updated the standard for piston-operated volumetric apparatuses (POVA) in 2022.

  • Download Guide: ISO 8655 Compliance - Key Points for Pipette Calibration

High-Throughput Experimentation (HTE)

Featured Device: XPR105MCP for Multichannel Pipettes

Watch to learn more about the XPR105MCP, which fulfills the ISO 8655 requirements for multichannel pipette calibration devices. This video covers the following points:

  • Increasing the speed of your multichannel pipette calibrations
  • Automating data collection and calculations with Calibry PC Software
  • Audit-readiness and certificate storage

Jump to one of the following sections to explore and learn more:

  1. Calibration of Pipettes Using a Balance – The Gravimetric Reference Method
  2. ISO 8655:2022, Important Changes for Pipette Calibration. Free Guide: ISO 8655 Compliance, Key Points for Pipette Calibration
  3. The Challenges of the Pipette Calibration Workflow
  4. Good Pipetting Practice™ (GPP™)
  5. Do You Need Support?
  6. Featured METTLER TOLEDO Solutions to Ensure Accurate Pipetting Results
    6.1Rainin Pipette Calibration Service
    6.2Calibration Balances and Routine Testing Devices
    • Routine Pipette Testing with SmartCheck™ – Fast and Easy Verification with Minimal Effort
    • Single-channel Pipette Testing with an XPR26PC Calibration Balance, for Volumes Down to 0.5 µL
    • Single-channel Pipette Testing with an XPR Analytical Balance and an Evaporation Trap
    • Multichannel Pipette Calibration with an XPR105MCP
    • Important Pipette Calibration Software and Accessories
  7. FAQ – Frequently Asked Questions about Pipette Calibration

Calibration of Pipettes Using a Balance – The Gravimetric Reference Method

The gravimetric reference measurement procedure for the accurate determination of volume is described in part 6 of ISO 8655:2022.

The balance used for testing shall be chosen according to the minimum requirements specified in ISO 8655-6:2022, Table 1, depending on the nominal volume of the apparatus under test. The balance parameters are defined so that the expanded uncertainty in use is less than one-fourth of the maximum permissible systematic error of the apparatus.

ISO 8655:2022 – Minimum Requirements for Balances

Nominal volume (V)

Resolution (mg)

Repeatability (mg)

Expanded uncertainty in use (mg)

0.5 μL ≤ V < 20 μL

0.001 *

0.01 **

0.006 *

0.03 **

0.012 *

0.06 **

20 μL ≤ V < 200 μL0.010.0250.05
200 μL ≤ V ≤ 10 mL0.10.20.4
10 mL < V ≤ 1,000 mL124
1,000 mL < V ≤ 2,000 mL101040

* Single-channel balance
** Multichannel balance, only valid for multichannel pipettes

To ensure reliable, accurate, traceable, and comparable results, all ISO 8655:2022 calibration methods (in 8655-6, 8655-7, and 8655-8) require a minimum of 10 repeated measurements per volume.

The calibration balance must not only meet the requirements as specified by ISO but must also undergo a program of calibration and routine testing — and tests must be up to date. Ensure your balances receive a calibration certificate that quantifies the measurement uncertainty in use according to EURAMET cg-18. This ensures the chosen balances are fit for the intended pipette calibration purpose and meet the new requirements of 8655-6. Ask METTLER TOLEDO for an Accuracy Calibration Certificate (ACC).

ISO 8655:2022, Important Changes for Pipette Calibration

After 20 years, the ISO 8655 standard was updated in May 2022. The changes in ISO 8655:2022 bring the standard up to date with current metrological principles to ensure accurate and reliable results.

Read more here about the updated ISO 8655:2022 requirements for pipettes:

  • Download Guide: ISO 8655 Compliance, Key Points for Pipette Calibration
Pipette Calibration Using a Balance (3)

The Challenges of the Pipette Calibration Workflow

It can be difficult to ensure an error-free pipette calibration workflow. Here are some of the key challenges faced by today’s labs:

  • Training technicians to have proper pipetting technique
  • Choosing the correct equipment for calibration
  • Ensuring a controlled environment, particularly for preventing evaporation and unstable readings
  • Completing calibrations under time pressure
  • Maintaining focus during repetitive and monotonous calibrations, particularly for multichannel pipettes
  • Transferring data without errors and correctly completing calculations
  • Maintaining organized document storage for audits
Pipette Calibration Using a Balance (4)

Good Pipetting Practice™ (GPP™)

Quality standards and guidelines, such as ISO 8655, ISO 17025, Good Laboratory Practice (GLP), and Good Manufacturing Practice (GMP), require regular calibration and/or verification of measuring instruments at specified intervals. This monitoring, calibration, control, and routine testing of instruments is intended to ensure metrological traceability and provide evidence of the pipette’s “fitness for purpose.”

METTLER TOLEDO’s Good Pipetting Practice™ (GPP™) program includes quality assurance measures for pipettes and simple, straightforward recommendations for their selection, installation, calibration, and operation. It provides a comprehensive, systematic approach to maximizing pipetting accuracy and reproducibility.

A combination of three complementary activities ensures that your pipette delivers reliable results every day:

  1. Pipette calibration and preventive maintenance – performed by a service provider
  2. Pipette routine testing – performed by the user
  3. Pipetting user training – delivered by a pipetting expert

Routine testing (also referred to as a pipette test, performance test, or quick check) provides reassurance that a pipette continuously delivers accurate and reliable volumes between calibrations. Compared to calibration, routine testing provides additional security because it checks the system of “user and pipette” under daily laboratory conditions, mimicking the actual pipetting process. This is particularly important for critical applications because the user technique can be a dominant source of pipetting uncertainty.

Pipette Calibration Using a Balance (5)

The type of sample, environmental conditions (humidity, temperature), SOPs, and user-defined tolerances determine to what extent and how frequently each activity should be performed. Good Pipetting Practice™ (GPP) supports you with creating a risk-based performance verification plan. Learn more about GPP™ and take an online pipetting risk check here: Good Pipetting Practice Risk Check

Pipette Calibration Using a Balance (6)

Do You Need Support?

Our team of experts in pipettes, balances, and calibration is happy to provide support. Please do not hesitate to contact us to discuss your requirements.

Information about GPP, pipette service intervals, or achieving ISO 8655:2022 compliance is available here: Services for Pipettes

For questions about pipette calibration balances, software, and accessories, please use the contact form below.

Pipette Calibration Using a Balance (7)

Rainin Pipette Calibration Service

Your work is too important to risk inaccurate or inconsistent data due to poor pipette performance. Rainin pipette service is rigorous and comprehensive, executed by highly trained and experienced technicians who know your pipettes. Make certain these important research tools deliver the reliability and reproducibility your work depends on with Rainin Service.

Reliable pipette service is part of the Rainin 360+™ system for optimal liquid handling results. Precision pipettes, high-purity tips, and expert service are all backed by innovative tools, software, and training to ensure exceptional performance across your workflows.

Whether you prefer mail-in or onsite pipette calibration, we offer a range of pipette calibrations that include ISO 8655 and ISO 17025-accredited pipette calibration to meet your regulatory needs.Rainin pipette service is rigorous and comprehensive, and it makes certain your pipettes deliver with the reliability and reproducibility your work depends on.

Simply put, when it comes to pipette service, no other provider can match the depth, breadth, and level of process integrity and control that METTLER TOLEDO Rainin offers.

Pipette Calibration Using a Balance (8)

Calibration Balances and Routine Testing Devices

To perform ISO 8655-compliant calibrations on a laboratory balance, METTLER TOLEDO Laboratory Weighing offers a comprehensive portfolio for the calibration of single and multichannel pipettes. Each balance can be connected with our Calibry PC Software, which guides technicians step-by-step through procedures and performs calculations automatically.

Devices for Pipette RoutineTesting and CalibrationSmartCheckXPR26PCXPR Balance with Evaporation TrapXPR105MCP

Pipette Type

Single-channel

Single-channel

Single-channel

Multichannel

Pipette Volume

10 µL -1,000 µL

0.5 µL -200 µL

0.5 µL -1,000 µL

0.5 µL -1,000 µL

User

Pipette user

Pipette users and calibration technicians

Pipette Test Type

Routine Testing

Routine testing and calibration

Frequency

1x/day – 1x/month

Routine testing: 1x/day – 1x/month, calibration: 1-4x/year (according to risk assessment)

Infrastructure

Lab bench

Dedicated weighing place (for routine testing) or controlled calibration room (for calibration)

Software

PipetteX

Calibry

Calibration Certificate

No

Yes

METTLER TOLEDO also providesbalance calibration services with an Accuracy Calibration Certificate (ACC). This ensures the chosen balance is fit for the intended pipette calibration purpose and meets the new requirements of 8655-6.

Pipette Calibration Using a Balance (9)

Routine Pipette Testing with SmartCheck – Fast and Easy Verification with Minimal Effort

With SmartCheck, you can test a pipette's accuracy between calibrations – right at your bench – in 60 seconds or less. Know that your pipettes are dispensing accurately and trust your data!

When used in conjunction with PipetteX™, the pipette testing data you collect using SmartCheck is automatically recorded into each pipette’s calibration and service record. PipetteX controls routine test methods, flags pipettes in the event of routine test failure, and sends notifications if action is needed. It reminds you when testing or calibration is due and supports compliance with 21 CFR Part 11.

Pipette Calibration Using a Balance (10)

Single-channel Pipette Testing with an XPR26PC Calibration Balance for volumes down to 0.5 µL

ISO 8655-6 requires that single-channel pipette volumes of less than 20 μL be measured on a six-place (microgram) balance. The XPR26PC was designed specifically for the measurement of these small volumes. The light barrier provides efficient hands-free operation with automatic opening and closing of the evaporation trap lid. Throughput is further improved by the large 10 mL waste container, which enables you to pipette continuously without interruption.

The supplementary Calibry PC software allows you to fulfill ISO 8655 and 21 CFR Part 11 conformity. Its simple user interface guides you through pipette calibration and testing procedures. Calibry stores all results in a secure database, informs you when upcoming tests are due, and generates customized reports.

Pipette Calibration Using a Balance (11)

Pipette Testing with an XPR Micro or Analytical Balance and an Evaporation Trap

An XPR Micro or Analytical Balance equipped with an evaporation trap is a perfect solution for calibrations and routine performance verifications of single-channel pipettes according to ISO 8655 requirements.
An XPR36/56 Microbalance with an evaporation trap fulfills ISO 8655requirements for pipettes with nominal volumes as low as 0.5 μL , while anXPR205 Analytical Balance with an evaporation trapis suitable for testing pipettes with volumes of 20 μL and greater.
The evaporation trap allows for higher throughput calibrations with fewer interruptions to empty the waste container. The supplementary Calibry PC software guides you through pipette calibration and testing procedures. It supports accurate and traceable pipette calibrations according to ISO 8655 and 21 CFR Part 11 requirements.

Pipette Calibration Using a Balance (12)

Multichannel Pipette Calibration with an XPR105MCP

The XPR105MCP is a high-speed solution to perform calibrations and routine performance verifications of multichannel pipettes with up to 12 channels according to ISO 8655 requirements. With a readability of 10 μg and repeatability of 15 μg, the system fulfills ISO 8655 requirements for multichannel pipettes with dispensing volumes as small as 0.5 μL.

The system has been carefully designed with a single load cell to ensure fast and equal performance for all channels. Up to 12 pipette channels can be measured in one dispense and the 360 measurements needed for a full calibration (12 channels, 3 ×10 repetitions) can be completed within 25 minutes. The load cell is protected from any leaks and spills to ensure lifelong performance.

The supplementary Calibry PC software guides you through pipette calibration and testing procedures. It supports accurate and traceable pipette calibrations according to ISO 8655 and 21 CFR Part 11 requirements.

Pipette Calibration Using a Balance (13)

Pipette Calibration Software and Accessories

Calibry PC softwareis ideal for managing ISO 8655 calibrations and routine tests of single and multichannel pipettes. Its simple user interface is easy to work with, and all data can be managed in compliance with 21 CFR Part 11. Calibry comes with pre-defined calibration methods for every pipette volume, or you can create your own custom test methods. It performs all calculations for efficient and seamless calibration processes, stores data from each pipette in a secure database, and generates customizable reports.

LabClimate PC software is an environmental measurement system consisting of sensors for temperature, pressure, humidity, and corresponding reporting software. The LabClimate software collects the data automatically and displays it clearly on a PC user interface. Data can also be synchronized directly with Calibry software or exported to Excel.

FAQ - Pipette Calibration

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Frequently Asked Questions on Pipette Calibration & ISO 8655

What is pipette calibration?

Pipette calibration is a process that determines the expected volume with the actual volume delivered by a pipette together with the associated measurement uncertainty by an accredited institution. The purpose is to prove if the pipette is still within its manufacture or ISO8655 specifications to deliver accurate results. Pipettes out of tolerance can often be serviced, adjusted and re-calibrated.

Why is pipette calibration important?

Pipette calibration shows the difference between the expected and actual volume, which shall be within a given tolerance. If it is outside the tolerance, it could potentially lead to incorrect analytical results.Consequently, experimental results would not be reproducible and entire analyses might need to be redone. To detect an out-of-tolerance pipette early, it is recommended that users check pipette accuracy at regular intervals between calibrations.

Why is it important to test pipettes in-between service intervals?

Up to 35% of pipettes sent to a service provider for calibration are out-of-specification. If your pipette is one of these, you may need to investigate which previous analyses could have been affected by this error. How can you avoid this in the future? Performing regular pipette quick checks uncovers any malfunctions early and eliminates surprises at your next calibration service.

What’s a quick way to check pipette accuracy?

Pipette accuracy shall be checked regularly between calibrations to avoid out-of-tolerance results between calibrations. Our SmartCheck device is designed specifically for quick tests—simply pipette 4 dispenses into the SmartCheck at your selected process tolerance, and the SmartCheck will indicate pass/fail results with a simple green/red light.

What are two procedures for calibrating multichannel pipettes?

There are two acceptable methods for calibrating and testing multichannel pipettes:

  1. Each channel can be measured individually, one after another, with a single-channel pipette calibration balance. For this purpose, test liquid is aspirated by all channels together and collected from one channel at a time.
  2. A multichannel pipette calibration balance can be used to measure the test volume delivered from all channels in parallel by aspirating and emptying each channel simultaneously and analyzing each channel’s results individually.
Pipette Calibration Using a Balance (2024)
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