Free Fd2400h3 - Federal

DEPARTMENT OF HEALTH AND HUMAN SERVICES PUBLIC HEALTH SERVICE FOOD AND DRUG ADMINISTRATION

LABORATORY

LOCATION

LAB #

MILK LABORATORY EVALUATION FORM
DATE X = DEVIATION O = NOT USED U = UNDETERMINED NA = NOT APPLICABLE

[Unless otherwise stated all tolerances are ±5%]
1. Laboratory Requirements (see CP, item 33) .............................. a. Unpreserved samples may be run up to 72 hours after initial collection ................................................................................. b. Samples may be run up to 7 days after initial collection if preserved with 0.02% 2-bromo-2-nitropropane-1,3-diol (BronopolTM)or 0.05% potassium dichromate (K2Cr2O7) ........ c. Comparative test with DMSCC ................................................. 1. Performed by each analyst performing ESCC test ............... 2. Test 4 samples (100K-200K, 300K-500K, 600K-800K and 900K-1.2M) in triplicate for both DMSCC (three separate smears each) and ESCC (three separate subsamples each, do not read same sample three times) ............................... 3. Results must be shown to be acceptable prior to official testing by analyst performing comparison, i.e. analyst is not certified until found acceptable. (co-requisite for certification) ...................................................................... 4. Copy of comparison and results in QC record (or easily accesible file in laboratory) ................................................. d. Analysts certified in DMSCC .................................................... Date Prep. ____________ Exp. Date ____________ ........ 2. Buffer/diluent Solution: Mix 0.5 L of stock solution with one bag (171 g) of Buffer 5000, add deionized water to 5 L, heat to 40 - 60C to speed process, store in diluent container next to instrument and use within 3 weeks (enough for about 10,000 samples) .................................. Date Prep. ____________ Exp. Date ____________ ........ 3. Rinse/sheath Liquid: Mix 100 mL of stock solution with deionized water to 20 L, store and use within 3 weeks (enough for about 2000 samples) ..................................... Date Prep. ____________ Exp. Date ____________ ........ 4. Insert Dye 5000 bag according to manufacturer's instructions .................................................................................... c. Stock Solutions for 5000 basic .............................................. 1. Dye/buffer stock solution: Dissolve 3 ethidium bromide tablets in 1 L of deionized water, stir to completely dissolve tablets, store in dark cool location, use within 16 weeks .... Date Prep. ____________ Exp. Date ____________ ........ 2. Clean 5000 stock solution: Dilute one bottle of Clean 5000 in 4.5 L of deionized water, heat to about 40 - 60C to speed process, store and use within 16 weeks ............. Date Prep. ____________ Exp. Date ____________ ........ d. Working Solutions for 5000 basic .......................................... 1. Buffer/diluent solution: Dissolve one bag of Buffer 5000 in about 8 L of deionized water in 10 L container, add 1 L of Clean 5000 stock solution and fill to a total of 10 L with deionized water, store and use within 6 weeks .................. Date Prep. ____________ Exp. Date ____________ ........ 2. Dye/buffer solution: Mix 1800 mL of Buffer/diluent solution and 200 mL of Dye stock solution in the reagent bottle and place in the instruments according to instructions, use within 6 weeks ...................................... Date Prep. ____________ Exp. Date ____________ ........ 3. Rinse/sheath liquid: Fill a 50 L container with about 49 L of deionized water, add 250 mL of Clean 5000 stock solution and fill to 50 L with deionize water, use within 3 weeks ..... Date Prep. ____________ Exp. Date ____________ ........ 5. Storage Periods and Temperatures .......................................... a. Storage time for Buffer 5000 and Clean 500 is one year from FE (Foss Electric, i.e., manufacture) date ............................... FE Date ____________ Exp. Date ____________ ................. b. Store Dye 5000 at 2-8C for a maximum of 1 year from FE Date FE Date ____________ Exp. Date ____________ ................. c. Rinse 5000, used to rinse dye interface when instrument has not been used for more than 7 days, store for 1 year from FE date, if seal is not broken, after seal is broken store at 2-8C for no more than 6 months .................................................... FE Date ____________ Exp. Date ____________ ................. 6. Other Liquids ...............................................................................
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EF

ELECTRONIC SOMATIC CELL COUNT Fossomatic 5000

APPARATUS
2. See Cultural Procedures, items 1-5 ........................................... 3. Water Bath .................................................................................. a. Circulating and thermostatically controlled to 37-42C ...........

REAGENTS
4. Preparation ................................................................................. a. 1 Liter bags ............................................................................. 1. Stock Solution: Dissolve 500 mL of Clean 5000 in 4.5 L of deionized water, heat to about 60C, store in airtight, light proof container in a cool location and use within 16 weeks .... Date Prep. ____________ Exp. Date ____________ ........ 2. Buffer/diluent Solution: Mix 1 L of stock solution with one bag (354 g) of Buffer 5000, add deionized water to 10 L, heat to 40 - 60C to speed process, store in diluent container next to instrument and use within 3 weeks (enough for about 25,000 samples) .................................................................. Date Prep. ____________ Exp. Date ____________ ........ 3. Rinse/sheath Liquid: Mix 250 mL of stock solution with deionized water to make 50 L, store and use within 3 weeks (enough for about 5000 samples) .......................... Date Prep. ____________ Exp. Date ____________ ........ 4. Insert Dye 5000 bag according to manufacturer's instructions .................................................................................... b. 0.5 Liter Bags .......................................................................... 1. Stock Solution: Dissolve 100 mL of Clean 5000 in 900 mL of deionized water (use syringe), heat to about 60C, store in airtight, light proof container in a cool location and use within 16 weeks ...................................................................
FORM FDA 2400h-3 (3/01)

LABORATORY

LAB #

LOCATION

DATE

ELECTRONIC SOMATIC CELL COUNT Fossomantic 5000
[Unless otherwise stated all tolerances are ±5%]
a. Blind solution: Prepare a 1% (w/v) NaC1 solution .................. b. Cleaning solution for milk waste tube: Prepare a 0.5% S-470 solution (5 g per Liter), heat to 50 - 70C, used to clean the milk waste tube at end of day, use within 1 week Date Prep. ____________ Exp. Date ____________ ............ c. Cleaning solution for flow cell flush: Use 0.5% S-470 solution (see item b above) ..................................................... 7. All solutions labeled with date prepared and expiration date ...... d. Hourly Control Sample (instrument drift check) ...................... 1. Use one of the standards (items 9a or b) in the 500-800K range, run in triplicate and determine average .................... 2. Optionally, prepare sufficient control/sample 500-800K range, run in triplicate and determine average ..................

PROCEDURE
10. Testing Standards (each time instrument used) ........................ a. Heat standards to 37-42C (using a temperature control) and read within 30 minutes of reaching temperature; use once and then discarded, i.e., do not re-use ........................................... b. Mix by inverting at least 2x, place in rack and put onto automatic track, run within 10 minutes .................................. c. Run the standards three times and average the counts for each level, records maintained ............................................... d. Each standard's average must be within 10% of the DMSCC (item 9) for that level, except within 15% for 100K-200K standard, records maintained ................................................. e. Repeatability - a standard in the 300K to 800K range must have a coefficient of variation (CV) of 5% or less on 10 replicates (Refer to Operating Manual), records maintained .... f. THESE PARAMETERS MUST BE ACHIEVED BEFORE PROCEEDING .......................................................................... 11. Testing Samples ......................................................................... a. Heat samples to 37-42C (using a temperature control) and read within 30 minutes of reaching temperature; samples must not be reused and must be discarded after use ............ b. Mix by inverting at least 2x, place in rack and put onto automatic track, run within 10 minutes of reaching the testing temperature ................................................................. 12. With continuous operation: ....................................................... a. Run a standard or optionally a control/sample (item 9d) in the 500K to 800K range hourly, must be within 5% of the original established instrument average value (optionally, within 10% of original DMSCC average) .................................................... b. Run control 3x ........................................................................ c. Run zero control (item 8d) ..................................................... d. Maintain records ..................................................................... 13. Routine maintenance ................................................................. a. Perform as described in operating manual ............................ b. Maintain records .....................................................................

START UP
8. Cell Counter ................................................................................ a. Turn power on and place instrument in standby mode .......... b. Check that enough rinse/sheath liquid, dye and buffer solutions are available in the external containers ................... c. Solutions not used beyond expiration date(s) ....................... d. Perform a blind check before starting measurement, if mean count is < 3000 cells/mL and individual measurements < 5000, within acceptable limits ............................................ e. IF ANY ABOVE PARAMETERS ARE WRONG, CORRECT BEFORE PROCEEDING ........................................................... f. Records maintained on all parameters each time instrument is used .................................................................................... 9. Milk Standards ........................................................................... a. Commercially prepared: ____________ Lot # ________ Date Rcvd. ____________ .......................... 1. Four standards in ranges 100K-200K, 300K-500K, 600K800K and 900K-1.2M ......................................................... 2. Do DMSCC in triplicate on each standard in set and average counts, records maintained .................................. 3. DMSCC check performed in rotation by all certified analysts .............................................................................. 4. Standards used within one week ........................................ b. Certified provider: _________________ Lot # ________ Exp. Date ____________ Date Rcvd. ____________ ...................................................... 1. Four standards in ranges 100K-200K, 300K-500K, 600K800K and 900K-1.2M ......................................................... 2. Maintain copies of all provided DMSCC values .................. 3. Measure and maintain records of temperature (0 - 7.2C) of standards as received .......................................................... 4. Maintain copies of all correspondence regarding problems .... c. Laboratory prepared (weekly) .................................................. 1. Prepare from raw milk > 18 hours old preserved with 0.05% potassium dichromate (K2Cr2O7) ............................. 2. Or, preserved with 0.02% 2-bromo-2-nitropropane-1,3diol (BronopolTM) ................................................................ 3. Standards cannot be preserved with formalin ..................... 4. Prepare 4 standards in ranges 100K-200K, 300K-500K, 600K-800K and 900K-1.2M, used within one week .......... Date prep. ____________ Exp. Date ____________ ...... 5. Do DMSCC in triplicate on each standard and average counts, records maintained ................................................ 6. DMSCC check performed in rotation by all certified analysts ...............................................................................
FORM FDA 2400h-3 (3/01)

REPORTS
14. Computing and Reporting Counts ............................................. a. Count obtained x 1000 is the cell count/mL milk ................... b. In reporting electronic somatic cell counts (ESCC/mL), record only first two left hand digits, raising second digit to next higher number when third digit is 6 or more ......................... c. Report the two left hand digits (rounded) .............................. 1. If the third digit is 5 the second digit is rounded by the following rule ..................................................................... a. When the second digit is odd round up, raise the second digit by 1 (odd up, 235 to 240) .........................

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LABORATORY

LAB #

LOCATION

DATE

ELECTRONIC SOMATIC CELL COUNT Fossomantic 5000
[Unless otherwise stated all tolerances are ±5%]
b. When the second digit is even round down, delete the 5 and report the second digit as is (even down, 225 to 220) ................................................................................ d. If count on instrument is < 100 report as < 100,000 ESCC/mL .................................................................................

FORM FDA 2400h-3 (3/01)

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