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Order Code PEWE Porphyrins Evaluation, Washed Erythrocytes

Reporting Name

Porphyrin Evaluation, RBC

Useful For

Preferred test for analysis of erythrocyte porphyrins

 

Establishing a biochemical diagnosis of erythropoietic protoporphyria, and X-linked dominant protoporphyria

Reflex Tests

Test ID Reporting Name Available Separately Always Performed
PPFWE Protoporphyrins, Fractionation, RBC Yes No

Testing Algorithm

This test is recommended for screening patients for possible erythropoietic protoporphyria and X-linked dominant protoporphyria. In addition, it can be used for evaluation of iron-deficiency anemia and chronic lead intoxication. Testing begins with total erythrocyte porphyrins. If the result is below 80 mcg/dL, it is normal, and testing is complete.

 

If the total erythrocyte porphyrin value is 80 mcg/dL or above, the protoporphyrin fractionation assay will automatically be performed at an additional charge. The fractionation test results include noncomplexed (free) protoporphyrin and zinc-complexed protoporphyrin.

 

The following algorithms are available:

-Porphyria (Acute) Testing Algorithm

-Porphyria (Cutaneous) Testing Algorithm

Performing Laboratory

Mayo Clinic Laboratories in Rochester

Specimen Type

Washed RBC


Ordering Guidance


This is the preferred test for assessment for protoporphyria. The preferred test for assessing lead toxicity in children is blood lead. For more information see PBDV / Lead, Venous, with Demographics, Blood or PBDC / Lead, Capillary, with Demographics, Blood. The preferred screening test for suspicion of a hepatic porphyria is urine porphyrins. For more information see PQNRU / Porphyrins, Quantitative, Random, Urine.



Necessary Information


1. Volume of packed cells and total volume of specimen (red cells + saline) are required and must be sent with specimen.

2. Include a list of medications the patient is currently taking.



Specimen Required


All porphyrin tests on erythrocytes can be performed on 1 draw tube.

 

Patient Preparation: Patient must not consume any alcohol for 24 hours before specimen collection.

Collection Container/Tube:

Preferred: Green top (sodium heparin)

Acceptable: Dark blue top (metal free heparin), green top (lithium heparin), or lavender top (EDTA)

Submission Container/Tube: Plastic vial

Specimen Volume: Entire washed erythrocyte suspension

Collection Instructions: Collect and process whole blood specimen as follows:

1. Transfer entire specimen to a 12-mL graduated centrifuge tube.

2. Centrifuge specimen at 4° C for 10 minutes at 2000 rpm.

3. Record volume of packed cells and the total volume of the specimen.

4. Discard supernatant plasma.

5. Wash packed erythrocytes 2 times by resuspension of at least an equal amount of cold 0.9% saline, mix, and centrifuge for 5 minutes at 2000 rpm, discarding supernatant after each washing.

6. Resuspend packed cells to the original total volume with 0.9% saline. Invert specimen gently to mix.

7. Transfer to a plastic tube and freeze.


Specimen Minimum Volume

1 mL of washed and resuspended erythrocytes

Specimen Stability Information

Specimen Type Temperature Time Special Container
Washed RBC Frozen (preferred) 14 days
  Refrigerated  14 days

Reference Values

PORPHYRINS, TOTAL, RBC

<80 mcg/dL

Day(s) Performed

Monday through Friday

CPT Code Information

84311-Spectrophotometry, analyte not elsewhere specified

82542-Chromatography (if appropriate)

Method Description

This evaluation is performed as a 2-step analysis. First, the total red blood cells (RBC) porphyrin concentration is determined by extracting the porphyrins from washed, resuspended RBCs using a mixture of ethyl acetate and acetic acid. The porphyrins are then back extracted into dilute hydrochloric acid. Total porphyrins are quantified using this extract via spectrofluorometry.(Piomelli S. Free erythrocyte porphyrins in the detection of undue absorption of Pb and Fe deficiency. Clin Chem. 1977;23:264-269; Gou EE, Balwani M, Bissell DM, et al. Pitfalls in erythrocyte protoporphyrin measurement for diagnosis and monitoring of protoporphyrias. Clin Chem. 2015;61[12]:1453-1456. doi:10.1373/clinchem.2015.245456)

 

If the total porphyrin concentration is elevated, the RBCs are re-extracted to separate and quantify the zinc-complexed and noncomplexed (free) protoporphyrin via high-performance liquid chromatography.(Smith RM, Doran D, Mazur M, Bush B. High-performance liquid chromatographic determination of protoporphyrin and zinc protoporphyrin in blood. J Chromatogr. 1980 Mar;181[3-4]:319-327; Gou EE, Balwani M, Bissell DM, et al. Pitfalls in erythrocyte protoporphyrin measurement for diagnosis and monitoring of protoporphyrias. Clin Chem. 2015;61[12]:1453-1456. doi:10.1373/clinchem.2015.245456)

Reject Due To

Cell suspension not available Reject

Method Name

Spectrofluorometric