US Environmental Protection Agency Office of Pesticide Programs Office of Pesticide Programs Mic ro bio lo gy La bo ra to ry Environmental Science Center, Ft. Meade, MD Standard Operating Procedure for Verification of Digital Pipettes SOP Number: QC-19-07 Date Revised: 09-26-14 ------- SOP No. QC-19-07 Date Revised 09-26-14 Page 1 of 8 SOP Number QC-19-07 Title Verification of Digital Pipettes Scope Describes process for verification of digital pipettes. Application Pipettes are calibrated annually and are evaluated using the gravimetric procedure as necessary. Approval Date SOP Developer: Print Name: SOP Reviewer Print Name: Quality Assurance Unit Print Name: Branch Chief Print Name: Date SOP issued: Controlled copy number: Date SOP withdrawn: ------- SOP No. QC-19-07 Date Revised 09-26-14 Page 2 of 8 TABLE OF CONTENTS Contents Page Number 1. DEFINITIONS 3 2. HEALTH AND SAFETY 3 3. PERSONNEL QUALIFICATIONS AND TRAINING 3 4. INSTRUMENT CALIBRATION 3 5. SAMPLE HANDLING AND STORAGE 3 6. QUALITY CONTROL 3 7. INTERFERENCES 3 8. NON-CONFORMING DATA 3 9. DATA MANAGEMENT 3 10. CAUTIONS 3 11. SPECIAL APPARATUS AND MATERIALS 3 12. PROCEDURE AND ANALYSIS 4 13. DATA ANALYSIS/CALCULATIONS 5 14. FORMS AND DATA SHEETS 5 15. REFERENCES 5 ------- SOP No. QC-19-07 Date Revised 09-26-14 Page 3 of 8 1. Definitions Abbreviations/definitions are provided in the text. 1. Verification failure = Verification of pipette exceeds ±5% of the target volume. 2. Ideal volume = Target volume; actual volume being measured, corrected for temperature using the density of water at 21.0°C (0.997995 g/mL, see section 15.1). 2. Health and Safety Follow procedures specified in SOP MB-01, Laboratory Biosafety. The Study Director and/or lead analyst should consult the Safety Data Sheet for specific hazards associated with products. 3. Personnel Qualifications and Training Refer to SOP ADM-04, OPP Microbiology Laboratory Training. 4. Instrument Calibration Refer to SOP EQ-03 (weigh balances) for details on method and frequency of calibration. 5. Sample Handling and Storage None. 6. Quality Control 1. For quality control purposes, the required information is documented on the appropriate form(s) (see section 14). 7. Interferences None. 8. Non- conforming Data 1. Management of non-conforming data will be specified in the study protocol; procedures will be consistent with SOP ADM-07, Non- Conformance Reports. 2. Do not use pipettes if the inaccuracy exceeds ±5% of the target volume. 9. Data Management 1. Data will be archived consistent with SOP ADM-03, Records and Archives. 2. Maintain an inventory of pipettes electronically using a Microsoft Excel spreadsheet (refer to section 14). After each addition to or deletion from the inventory, file a hard copy of the pipette inventory in the Pipette Verification and Calibration Record Book. 10. Cautions 1. If a pipette is dropped or damaged, it must be successfully verified using the gravimetric procedure or recalibrated by a vendor prior to use. 2. If a pipette fails an in-house verification assessment, it will not be used in the laboratory and will be recalibrated by a vendor prior to use. 11. Special Apparatus and 1. Calibrated balances. Capable of measuring 0.01 g for verifying pipettes with volumes greater than or equal to 1 mL and 0.001 g for verifying ------- SOP No. QC-19-07 Date Revised 09-26-14 Page 4 of 8 Materials pipettes with volumes less than 1 mL. 2. Pipettes. a. Rainin Adjustable Volume Pipettes b. Gilson Microman Positive Displacement Pipettes c. Gilson Distriman Continuously Adjustable Volume Repetitive Pipette 12. Procedure and Analysis 12.1 Pipette Verification Requirements a. Annually verify and service pipettes using an ISO accredited vendor. b. If a pipette is dropped or broken, verify the pipette using the gravimetric verification procedure. c. Record the annual verification results and if necessary, gravimetric analysis results on the Pipette Verification Record Sheet (refer to section 14). The Pipette Verification Record Sheet is based on the inventory of pipettes and may change over time. 12.2 Gravimetric Verification Procedure e. f. Record all pertinent information for the gravimetric verification procedure on the Pipette Verification - Gravimetric Analysis Form (refer to section 14). In advance of testing, fill a container with de-ionized water and allow it to equilibrate to room temperature in the same laboratory with the balance. List the balance that will be used on the appropriate form. Place a small Erlenmeyer flask on the balance and record its mass on the appropriate form. Using the pipette to be verified, aspirate an aliquot of DI water from the sample aliquot container and dispense into the Erlenmeyer flask. Record the mass on the appropriate form. Follow the procedure in 12.2d-e for each subsequent sample addition; measure at least 5 samples. Do not tare between samples. After each new sample addition, record the mass on the appropriate form. Input the measurements and other appropriate information into the Pipette Verification - Gravimetric Analysis Spreadsheet. Verify that the percent inaccuracy is within ±5%. If the percent inaccuracy is outside of this range, remove the pipette from use until it is repaired and calibrated by a vendor. ------- SOP No. QC-19-07 Date Revised 09-26-14 Page 5 of 8 „ , . (x-ideal volume)x 100 i. Percent Inaccuracy = - ideal volume 12.3 Pipette Verification Record Sheet a. From the data on the calibration certificate for each pipette, record the following on the Pipette Verification Record Sheet: i. Verification Status: record as "pass" or "fail". Record status as "pass" if the percent inaccuracy from gravimetric analysis is within ±5% or if vendor calibration certificate indicates acceptable results, otherwise record as "fail". ii. Corrective Action: record as "yes" or "no". Record as "yes" if the verification status is "fail" and describe the action taken at the bottom of the page. If the verification status is "pass," record as "no." b. Generate one Pipette Verification Record Sheet annually and file in the Pipette Verification and Calibration Log Book. c. The Pipette Verification Record Sheet may be completed electronically or by hand. 13. Data Analysis/ Calculations 1. None 14. Forms and Data Sheets 1. Sample Pipette Verification Record Sheet 2. Sample Pipette Inventory 3. Test Sheets. Test sheets are stored separately from the SOP under the following file names: MLB Pipette Inventory QC-19-07 Fl.xlsx Pipette Verification Record Sheet QC-19-07_F2.xlsx Pipette Verification - Gravimetric Analysis Form QC-19-07 F3.xlsx Pipette Verification - Gravimetric Analysis ir. 1 c , , A QC-19-07 F4.xlsx Spreadsheet - 15. References 1. CRC Handbook of Chemistry and Physics. 93rd ed. CRC Press: Boca Raton, FL, 2012; p 6-8. ------- SOP No. QC-19-07 Date Revised 09-26-14 Page 6 of 8 Sample Pipette Verification Record Sheet OPP Microbiology Laboratory Verification Date(s): Initials: Manufacturer Model No. Serial No(s). Volume Range In-house Verification Volume(s) Verification Status1 Corrective Action (Y/N) X12523D Gilson M10 X12649D 1-10 nL 10 nL BH15232 Gilson M100 GG05125 10-100 nL 100 nL GG05127 AE10020 10 nL (1-12.5 nL) Gilson Distriman BD10010 1-1250 nL 100 (iL (10-125 (iL) 900 nL (100-1250 HL) U10048H A0504243A L0508039A C0823986A J0753884A C0825980A Rainin L-1000 C0823596A 100- 1000 nL H0101474A 1000 nL H0100977A J0902334A J0908624A D0303509A E0301364A A0510192A L0509218A C0821962A J0750805A Rainin L-200 C0820542A 20-200 nL 100 nL C0820661A G0101809A G0102379A J0902612A C0401654A ------- SOP No. QC-19-07 Date Revised 09-26-14 Page 7 of 8 Manufacturer Model No. Serial No(s). Volume Range In-house Verification Volume(s) Verification Status1 Corrective Action (Y/N) Rainin L-100 C0825210A 10-100 nL 100 nL C0825238A A0507382A C0822081A J0724932A C0822312A Rainin L-20 C0825315A 2-20 nL 10 nL F0100492A F0100448A J0903005A C0400398A Rainin L-2 H0100116A 0.1-2 nL 2 nL H0100003A Rainin L-5000 D1080497A 0.5-5 mL 5 mL Rainin L-10000 A1058487A 1-10 mL 10 mL Rainin L-20000 L0931886A 2-20 mL 20 mL Verification status = PASS if percent inaccuracy from gravimetric analysis is within ±5% or if vendor calibration certificate indicates acceptable results. If verification status = FAIL, record "yes" in Corrective Action column and fill in Action Taken below. Corrective Actions: Pipette Serial Number Action Taken 1) 2.) ------- SOP No. QC-19-07 Date Revised 09-26-14 Page 8 of 8 Sample Pipette Inventory OPP Microbiology Laboratory MLB Pipette Inventory Manufacturer Modi! No Serial Xt/i":). X12523D Gils on mm X12649D BK15232 Gils on mm GG0512" GG05125 AE10020 Gils on Distriman BD10Q13 UIOCiSH A050-243A LQ53S93SA CQS239SSA J0753SS4A C0S25980A Raintn L-1000 C5S23536A HOIOU7iA HC10G9-W JO'90233-A IMDS524A DC333309A E33C135-A Manufacturer Model Mo. S^rjai Xstit A0310I92A L350S21SA CCS21P52A J37J0S35A Ramin L-2Q0 C0820542A C3S23S61A G31G1S3SA GO 1323^3A J09C2522A C340165-A Rainin 1-100 CGS252I3A C3S2523SA A0W3S2A C0S223S1A J372-1932A C0S223I2A Ramm L-20 008253 ISA F013012SA F013C-USA J39C3G35A CD-3C39SA Rainin L-2 FECI 03116A K01300C3A Rainin L-50'03 D10SD-t9~A Rainin L-20003 AIDSS-S'A Raiiiin L-23CC0 L0931SS6A ------- |