The Water Boiling Test (WBT ) with ISO/IWA Metrics and ARC Equipment
Volumetric performance assessment of automated liquid ......Introduction: In 2015, the International...
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Introducon: In 2015, the Internaonal Organizaon for Standardizaon (ISO) published a new document, ISO IWA 15, which describes standardized ways of determining and reporng the volumetric performance of automated liquid handling systems (ALHS). A. Bjoern Carle, PhD George Rodrigues, PhD Volumetric performance assessment of automated liquid handling systems with ISO IWA 15 25 Bradley Drive, Westbrook, Maine 04092 888-406-3463 | [email protected] | © 2016 Artel, Inc. Figure 1: While ISO standards exist for the calibraon of volumetric glassware (ISO 4787) and handheld pipees (ISO 8655), ISO IWA 15 addresses the need for a global document addressing volumetric performance of automated liquid handling systems. mL µL nL Mechanical single and multi-channel pipettes Acoustic dispensing systems Automated pipetting and dispensing systems Picoliter dispensing systems pL ISO 4787 ISO 8655 ISO IWA 15 Evolution of Liquid Handling Technology Volumetric and Mohr pipettes (Class A glassware) 1900’s 2015 Figure 2: Standardized volumetric performance assessment improves harmony across liquid handling plaorms and geographic regions. Acknowledgements The following organizaons parcipated in development of ISO IWA 15: Agilent Technologies, Inc. Analyk Jena AG Artel, Inc. Brand GmbH & Co. KG Eppendorf AG Gilson, Inc. Hamilton Bonaduz AG Integra Biosciences AG Meler-Toledo Ltd. Sartorius AG Tecan Schweiz AG Thermo Fisher Scienfic Oy University of Freiburg (IMTEK) Review and helpful suggesons were provided by: Brish Columbia Genome Sciences Center Hinsdale Pathology Associates Merck Research Laboratories Naonal Virus Reference Laboratory, University College of Dublin North Carolina State University (BETEC) Contents of ISO IWA 15 Terminology: 50 definions Operaon of ALHS: Environmental condions, acceptance tesng, user tesng Volumetric Performance: Indexing to track data, descripve stascs Test Methods: Photometric, weighing, and other methods Specificaons: Informaon to be provided as part of performance specificaon Reporng of Results: Test reports and calibraon cerficates Applicaons of Descripve Stascs: Four examples Bibliography: 41 references ISO IWA 15 consensus document developed under the direction of ISO/TC 48 Publish ISO IWA 15 Future work proposed by participants ISO/TC 48/WG 04 to lead suggested technical projects Collaboration with measurement research groups and projects Series of ISO standards to be developed Need for ALHS standardization identified ISO IWA 15 project initiated Example 1 8 channel device, L =8 one single run, M =1 12 replicates per channel, N = 12 measured in one 96 well plate Form. 1 Form. 3 Form. 7 Channel 1 2 3 4 5 6 7 8 9 10 11 12 V(l) e S (l) CV(l ) l =1 A 97.87 97.72 98.33 98.99 98.79 97.98 98.43 99.32 95.69 99.46 99.23 97.59 98.28 1.72% 1.06% l =2 B 97.60 99.53 98.27 99.98 98.44 100.42 100.28 98.54 97.37 100.84 99.95 102.07 99.44 0.56% 1.42% l =3 C 98.23 96.62 95.95 96.14 98.24 96.79 97.77 97.93 98.31 96.28 96.99 97.53 97.23 2.77% 0.90% l =4 D 102.49 99.09 100.74 100.41 101.02 100.15 100.12 100.53 102.11 99.94 99.19 102.36 100.68 0.68% 1.13% l =5 E 97.31 96.41 98.07 97.97 96.73 99.27 99.13 98.58 96.93 98.30 97.35 98.31 97.86 2.14% 0.94% l =6 F 96.53 96.35 97.54 95.95 95.55 97.79 97.99 97.48 98.87 98.62 97.42 97.65 97.31 2.69% 1.05% l =7 G 97.11 97.13 99.12 97.16 96.53 97.10 98.15 97.14 99.45 97.33 98.16 98.05 97.70 2.30% 0.91% l =8 H 97.86 95.60 98.50 99.43 98.56 98.37 97.11 98.53 97.35 97.47 97.78 97.57 97.84 2.16% 0.99% max e S (l) 0.68% Run Order Statistics min e S (l) 2.77% n → 1 2 3 4 5 6 7 8 9 10 11 12 max CV(l) 1.42% Form. 9 V(n) 98.13 97.30 98.32 98.25 97.98 98.48 98.62 98.50 98.26 98.53 98.26 98.89 Form. 3 e S (n) 1.87% 2.70% 1.68% 1.75% 2.02% 1.52% 1.38% 1.50% 1.74% 1.47% 1.74% 1.11% Form. 11 CV (n) 1.88% 1.42% 1.38% 1.75% 1.73% 1.36% 1.14% 1.09% 1.99% 1.53% 1.09% 2.09% Form. 14 Grand Avg Form. 12 CV C2C V GA 98.29 1.21% Overall CV Form. 13 CV OA 1.53% Channel Statistics Overall Statistics Figure 3: Accurate results are achieved by improving both precision and trueness . Improving Precision Decreasing Random Errors Improving Trueness Decreasing Systematic Errors Example 1: Test design and stascal calculaons In this example an 8 channel liquid handling system is used to fill a 96 well plate with 100 µL per well. The delivered volume is measured in each well, and descripve stascs are calculated. The Formula references are highlighted in green (e.g., Form. 14) and each formula is described in detail within ISO IWA 15. This figure corresponds to Figure A.4 in ISO IWA 15. Copyright remains with ISO. ISO IWA 15 applies current definions from the internaonal vocabulary of metrology (VIM) and adapts them for the specific case of automated liquid handling systems. This figure shows the relaonships between three conceptual terms (accuracy, precision and trueness) along with quanfiable values (uncertainty, random error and systemac error). Figure 3 is adapted from ISO IWA 15, and a descripon of all 50 terms is contained in that document. Copyright remains with ISO. Example 2: Updated terminology Future Work Literature Cited 1. ISO 4787:2010 Laboratory glassware - Volumetric instruments - Methods for tesng of capacity and for use. 2. ISO 8655 1:2002, Piston-operated volumetric apparatus - Parts 1 - 7. 3. ISO IWA 15:2015 Specificaon and method for the determinaon of volumetric performance of automated liquid handling systems. Contact Informaon The authors welcome inquiries and can be reached directly at the following email addresses: Dr. George Rodrigues: [email protected] Dr. A. Bjoern Carle: [email protected] These three documents are available for purchase at the ISO webstore www.ISO.org/ISO/store.htm artel-usa.com
Transcript of Volumetric performance assessment of automated liquid ......Introduction: In 2015, the International...
Introduction: In 2015, the International Organization for Standardization (ISO) published a new document, ISO IWA 15, which describes standardized ways of determining and reporting the volumetric performance of automated liquid handling systems (ALHS).
A. Bjoern Carle, PhDGeorge Rodrigues, PhD
Volumetric performance assessment of automated liquid handling systems with ISO IWA 15
25 Bradley Drive, Westbrook, Maine 04092888-406-3463 | [email protected] | © 2016 Artel, Inc.
Figure 1: While ISO standards exist for the calibration of volumetric glassware (ISO 4787) and handheld pipettes (ISO 8655), ISO IWA 15 addresses the need for a global document addressing volumetric performance of automated liquid handling systems.
mLµL
nL
Mechanicalsingle and multi-channel
pipettes
Acoustic dispensing systems
Automated pipetting and dispensing systems
Picoliter dispensing systems
pL
ISO 4787 ISO 8655 ISO IWA 15
Evol
utio
n of
Liq
uid
Han
dlin
g Te
chno
logy
Volumetric and Mohr pipettes (Class A glassware)
1900’s 2015
Figure 2: Standardized volumetric performance assessment improves harmony across liquid handling platforms and geographic regions.
Acknowledgements The following organizations participated in development of ISO IWA 15:Agilent Technologies, Inc.Analytik Jena AGArtel, Inc.Brand GmbH & Co. KGEppendorf AGGilson, Inc.Hamilton Bonaduz AGIntegra Biosciences AGMettler-Toledo Ltd.Sartorius AGTecan Schweiz AGThermo Fisher Scientific OyUniversity of Freiburg (IMTEK)
Review and helpful suggestions were provided by:British Columbia Genome Sciences CenterHinsdale Pathology AssociatesMerck Research LaboratoriesNational Virus Reference Laboratory, University College of DublinNorth Carolina State University (BETEC)
Contents of ISO IWA 15 Terminology: 50 definitionsOperation of ALHS: Environmental conditions, acceptance testing, user testing Volumetric Performance: Indexing to track data, descriptive statistics
Test Methods: Photometric, weighing, and other methodsSpecifications: Information to be provided as part of performance specification
Reporting of Results: Test reports and calibration certificates Applications of Descriptive Statistics: Four examplesBibliography: 41 references
ISO IWA 15 consensus document developed under the direction of ISO/TC 48
Publish ISO IWA 15
Future work proposed by participants
ISO/TC 48/WG 04 to lead suggestedtechnical projects
Collaboration with measurement research groups
and projects
Series of ISO standards to be developed
Need for ALHS standardization identifiedISO IWA 15 project initiated
Example 18 channel device, L = 8one single run, M = 112 replicates per channel, N = 12measured in one 96 well plate
Form. 1 Form. 3 Form. 7Channel 1 2 3 4 5 6 7 8 9 10 11 12 V(l) e S(l) CV(l)
l = 1 A 97.87 97.72 98.33 98.99 98.79 97.98 98.43 99.32 95.69 99.46 99.23 97.59 98.28 -‐‑1.72% 1.06%l = 2 B 97.60 99.53 98.27 99.98 98.44 100.42 100.28 98.54 97.37 100.84 99.95 102.07 99.44 -‐‑0.56% 1.42%l = 3 C 98.23 96.62 95.95 96.14 98.24 96.79 97.77 97.93 98.31 96.28 96.99 97.53 97.23 -‐‑2.77% 0.90%l = 4 D 102.49 99.09 100.74 100.41 101.02 100.15 100.12 100.53 102.11 99.94 99.19 102.36 100.68 0.68% 1.13%l = 5 E 97.31 96.41 98.07 97.97 96.73 99.27 99.13 98.58 96.93 98.30 97.35 98.31 97.86 -‐‑2.14% 0.94%l = 6 F 96.53 96.35 97.54 95.95 95.55 97.79 97.99 97.48 98.87 98.62 97.42 97.65 97.31 -‐‑2.69% 1.05%l = 7 G 97.11 97.13 99.12 97.16 96.53 97.10 98.15 97.14 99.45 97.33 98.16 98.05 97.70 -‐‑2.30% 0.91%l = 8 H 97.86 95.60 98.50 99.43 98.56 98.37 97.11 98.53 97.35 97.47 97.78 97.57 97.84 -‐‑2.16% 0.99%
max e S(l) 0.68%
Run Order Statistics min e S(l) -‐‑2.77%
n → 1 2 3 4 5 6 7 8 9 10 11 12 max CV(l) 1.42%
Form. 9 V(n) 98.13 97.30 98.32 98.25 97.98 98.48 98.62 98.50 98.26 98.53 98.26 98.89
Form. 3 e S(n) -‐‑1.87% -‐‑2.70% -‐‑1.68% -‐‑1.75% -‐‑2.02% -‐‑1.52% -‐‑1.38% -‐‑1.50% -‐‑1.74% -‐‑1.47% -‐‑1.74% -‐‑1.11%
Form. 11 CV(n) 1.88% 1.42% 1.38% 1.75% 1.73% 1.36% 1.14% 1.09% 1.99% 1.53% 1.09% 2.09%
Form. 14
Grand Avg Form. 12 CVC2C
V GA 98.29 1.21%
Overall CV Form. 13
CVOA 1.53%
Figure A.4
Channel Statistics
Overall Statistics
Figure 3: Accurate results are achieved by improving both precision and trueness .
Improving AccuracyDecreasing Uncertainty
Improving PrecisionDecreasing Random Errors
Impr
ovin
g Tr
uene
ssD
ecre
asin
g Sy
stem
atic
Err
ors
Example 1: Test design and statistical calculations In this example an 8 channel liquid handling system is used to fill a 96 well plate with 100 µL per well. The delivered volume is measured in each well, and descriptive statistics are calculated. The Formula references are highlighted in green (e.g., Form. 14) and each formula is described in detail within ISO IWA 15. This figure corresponds to Figure A.4 in ISO IWA 15. Copyright remains with ISO.
ISO IWA 15 applies current definitions from the international vocabulary of metrology (VIM) and adapts them for the specific case of automated liquid handling systems. This figure shows the relationships between three conceptual terms (accuracy, precision and trueness) along with quantifiable values (uncertainty, random error and systematic error). Figure 3 is adapted from ISO IWA 15, and a description of all 50 terms is contained in that document. Copyright remains with ISO.
Example 2: Updated terminology Future Work
Literature Cited1. ISO 4787:2010 Laboratory glassware - Volumetric instruments - Methods for testing of capacity and for use.
2. ISO 8655 1:2002, Piston-operated volumetric apparatus - Parts 1 - 7.
3. ISO IWA 15:2015 Specification and method for the determination of volumetric performance of automated liquid handling systems.
Contact InformationThe authors welcome inquiries and can be reached directly at the following email addresses:Dr. George Rodrigues: [email protected]. A. Bjoern Carle: [email protected]
These three documents are available for purchase at the ISO webstore www.ISO.org/ISO/store.htm
artel-usa.com
