Biopro J Met Phosphorus 508
Biopro J Met Phosphorus 508
Biopro J Met Phosphorus 508
Analyte: Phosphorous
Matrix: Serum
Method No.:
Revised:
The Advanced Research and Diagnostic Laboratory (ARDL) periodically refine these
laboratory methods. It is the responsibility of the user to contact the person listed on the title
page of each write-up before using the analytical method to find out whether any changes
have been made and what revisions, if any, have been incorporated.
Phosphorous
NHANES 2017-2018
This document details the Lab Protocol for testing the items listed in the following table:
1 of 14
Phosphorous
NHANES 2017-2018
Eighty percent of phosphorous in the human body is located in the bone in the form of
calcium phosphate as the apatite, Ca2+(Ca3(PO4)2)32-. The remainder is involved in
intermediary carbohydrate metabolism, and in phospholipids, nucleic acids and ATP.
Phosphorous occurs in the blood as inorganic phosphate and in organically bound
phosphoric acid.
2. SAFETY PRECAUTIONS
Caution: This product is of human and animal origin. Handle as though capable of
transmitting infectious disease. Wear appropriate PPE when handling equipment,
reagents, and samples. This kit contains components classified as follows according to
the European Directive 1999/45/EEC: C - Corrosive (Sulfuric acid) R 35; S 26, S
36/37/39, S 45 Causes severe burns. In case of contact with eyes, rinse immediately
with plenty of water and seek medical advice. Wear suitable protective clothing, gloves
and eye/face protection. In case of accident or if you feel unwell, seek medical advice
immediately (show the label where possible).
2 of 14
Phosphorous
NHANES 2017-2018
3 of 14
Phosphorous
NHANES 2017-2018
f. Specimen Handling and Transport: Mix specimens well, allow clot to fully form (if
serum), and centrifuge 10 minutes at 2000 x g before use. Aliquot a minimum of 0.1
mL. Store sample in refrigerator until shipment. Ship at refrigerated temperature.
•R2 reagent. Ammonium molybdate: 3.5 mmol/L; sulfuric acid: 0.36 mol/L;
sodium chloride: 150 mmol/L
•Storage and stability. Keep reagents refrigerated until use. The reagents are
stable for 12 weeks refrigerated on the analyzer.
c. Equipment/Instrumentation-
e. Quality Control
4 of 14
Phosphorous
NHANES 2017-2018
5 of 14
Phosphorous
NHANES 2017-2018
b. Professional Judgement: Check results for error flags and take appropriate
corrective action. Investigate alert values and delta checks.
c. Result Entry
Manual entry
• Report low results as <0.3 mg/dL for serum or <3 mg/dL for urine.
6 of 14
Phosphorous
NHANES 2017-2018
• Check results for error flags and take appropriate corrective action.
Out of Range results: Certain tests have pre-programmed limits that trigger an automatic
re-analysis by the COBAS. These limits may be low-end values or high-end values (but
within technical range). If the duplicate value is in agreement with the initial value, then the
initial value is reported.
Results are reported to one decimal places (as x.x) in mg/dL. Report low results as <0.3
mg/dL.
Dilutions: The confirmed analytical measurement range of the assay is 0.3-20 mg/dL for
serum, and 3-285 mg/dL for urine. Specimens exceeding the high limit are automatically
diluted (1:2) by the instrument. Results from samples diluted using the rerun function are
automatically multiplied by 2. If a manual dilution is required, dilute the specimen in normal
saline, and multiply the result by the dilution factor. For example, to perform a 1:5 dilution,
pipette 50 μL of the patient sample into 200 μL of normal saline. Mix thoroughly, perform
the assay, and multiply the result by a factor of 5. The maximum allowable manual dilution
is 1:5.
7 of 14
Phosphorous
NHANES 2017-2018
•Normal pooled serum control (CQ). Stable at -80°C for up to 4 years, at refrigerated
temperature for up to 1 day and at room temperature for up to 4 hours.
•Both levels of quality control are analyzed at the start of the day and results are
verified for acceptability prior to testing specimens. Quality control is also analyzed
at the end of the shift, with change in reagent, after major maintenance, or as
needed for troubleshooting.
•New Lot Verification: Each new reagent lot must be verified for acceptability before
being placed into use. Calibration, quality control, and comparison of at least 5
patient samples on the old and new lots must be performed and found to be within
acceptable limits before a new lot can be placed into use.
8 of 14
Phosphorous
NHANES 2017-2018
2. Perform a calibration.
4. Contact Roche Technical Support for assistance and possible service dispatch.
Phone: 1-800-428-2336; account number: 55042919
Not applicable.
Specimens are stored at refrigerated temperature between sample receipt and analysis
on the instrument. Specimens must be at room temperature prior to assay. Specimens
are returned to refrigerated or frozen temperature post analysis depending on the study
specific requirements.
9 of 14
Phosphorous
NHANES 2017-2018
Should the testing system become inoperable, discontinue testing and notify the
supervisor. While instrument trouble-shooting or repair occurs; keep specimens at
refrigerated or frozen temperature depending on study specific requirements.
All data is reported electronically via an eFile that is uploaded to the WESTAT secure
website within 21 days of receipt of specimens.
Specimen Receipt:
Shipments for NHANES generally will arrive on Tuesdays and/or Wednesdays. These
shipments are recorded on the Log of Quality Assurance located in a binder labeled
NHANES Shipping Log in the receiving area. The specimen barcode numbers in the
boxes are checked against the manifests. The receipt date is written on top of the
boxes. The frozen samples (vial 11-Iron/UIBC & vial 13-CRP) are placed in the
designated -70°C freezer and the refrigerated samples (vial 18-Biochem panel) are
placed in the designated 2-8°C refrigerator until analysis. The manifests are filed in a
binder labeled NHANES Shipping Manifests located in the receiving area. All labels are
removed from the shipping box and the provided airbill is attached for return shipment.
10 of 14
Phosphorous
NHANES 2017-2018
Specimen Ordering/Labeling:
Electronic files for all NHANES specimens are sent via email from Westat, Inc to the
NHANES contact person shortly before they are to be received. These files include the
Sample ID, Analyte Type, Slot No, Sample Collection Date, Sample Comment, Age
Grouping, Astro ID, Receipt Date, Analysis Date, Run Number, Tech ID, Analyte Result,
Result Comment, Adjusted Result, QC Repeat, LOD, Change Reason, and Change
Reason Other. The first seven columns are protected and cannot be altered. The files
are saved on the laboratory’s common S drive in the NHANES Biochem folder. After
analysis, the contact person returns the completed files via their website to Westat, Inc.
11 of 14
Phosphorous
NHANES 2017-2018
The NHANES spreadsheets are used to set up pending batches for batch accession
upload in the Laboratory Information system (STARLIMs). New labels are generated out
of the Laboratory Information System (STARLIMs). The new bar-coded labels are
attached to a carrier tube. The Cobas analyzer reads the bar-coded label for the sample
ID and test information.
Specimen Storage:
The temperatures for all freezers and refrigerators are monitored 24 hours a day/ 7 days
a week. If the temperature for any unit falls outside the allowable range, action is taken
to resolve the problem. If the temperature cannot be corrected, the contents are moved
to a different unit.
Prior to analysis, the specimens are stored in the designated 2-8°C refrigerator. On the
day of analysis, the specimens are selected and thawed by the technician operating the
COBAS. After analysis and the QC repeats have been run, the specimens are refrozen.
After 1 year, the specimen vials that have at least 0.2ml of sample remaining will be
shipped to SriSai Biopharmaceuticals in Frederick, MD. These specimens will be
shipped on dry ice via Federal Express.
12 of 14
Phosphorous
NHANES 2017-2018
13 of 14
Phosphorous
NHANES 2017-2018
REFERENCES
1. Cobas 6000 Analyzer Operator’s Manual. Roche Diagnostics Inc., 9115 Hague Road
Indianapolis, IN 46250-0457. Version 5.0.
2. Roche Calibrator for Automated Systems (C.F.A.S.) package insert. Version 2011-
08, V6. Roche Diagnostics Inc., 9115 Hague Road Indianapolis, IN 46250-0457.
3. Roche PHOS2 reagent package insert. Version 2010-10, V5. Roche Diagnostics
Inc., 9115 Hague Road Indianapolis, IN 46250-0457.
4. Roche Precipath U Plus package insert, Version 2011-04, V4. Roche Diagnostics
Inc., 9115 Hague Road Indianapolis, IN 46250-0457.
14 of 14