Effect of in vitro hemolysis on chemical values for serum.

Author(s): Frank JJ, Bermes EW, Bickel MJ, Watkins BF

Publication: Clin Chem, 1978, Vol. 24, Page 1966-70

PubMed ID: 709831 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of hemolysate concentration on measured clinical chemistry analytes in serum measured by several methods. Erythrocytes obtained from EDTA-whole blood and lysed by the addition of de-ionized water and sodium saponin were added to thawed pools of serum. Serum containing hemolysate was preserved with disodium citrate monohydrate and stored at -20 degrees C for less than 24 h before analysis for all analytes except acid phosphatases, which were assayed within 48 h.

   Summary of Findings:

   For all measurement methods, increasing hemolysate concentration resulted in increases in aspartate aminotransferase (66-84%), LDH (137-155%), alpha-hydroxybutyrate dehydrogenase (153%), CK (35-130%), acid phosphatase (15-315%), potassium (16-19%) and iron (10-24%) and decreases in bilirubin (-28%) and bicarbonate (-7.5%) levels . However, uric acid, urea nitrogen, albumin, alkaline phosphatase, cholesterol, total protein, sodium, magnesium, calcium, inorganic phosphorous and chloride were unaffected by hemolysate concentration. With the exception of CK,acid phosphatase and iron, the magnitude of changes observed was similar among methods. The magnitude of the increase in CK with the addition of 2800 mg/L hemoglobin was much greater when measured using the DuPont aca (130%) than when measured using the Gilford 3500 with Worthington reagents (35%). The magnitude of the increase in acid phosphatase levels with the addition of 2800 mg/L hemoglobin was much greater when measured using Bessey, Lowry and Brock method (315%) than when measured using the Bodansky method (15%) or the DuPont aca (22%). Similarly, while only a 10% increase in iron with the addition of 2800 mg/L hemoglobin was identified by the DuPont aca method a 24% increase was identified using the Henry method.

   Biospecimens

   • Bodily Fluid - Serum

   Preservative Types

   • Frozen

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   Carbohydrate	Clinical chemistry/auto analyzer
   Protein	Clinical chemistry/auto analyzer
   Small molecule	Clinical chemistry/auto analyzer
   Steroid	Clinical chemistry/auto analyzer
   Electrolyte/Metal	Ion selective electrode
   Electrolyte/Metal	Flame emission photometry
   Electrolyte/Metal	Atomic absorption spectroscopy
   Electrolyte/Metal	Clinical chemistry/auto analyzer

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Biospecimen components	Less than 90 mg hemoglobin/L 320 mg hemoglobin/L 1440 mg hemoglobin/L More than 2800 mg hemoglobin/L
   Clinical chemistry/auto analyzer Specific	Technology platform	DuPont aca Henry method Gilford 3500 using Dade reagents Gilford 3500 using Worthington reagents Bodansky Bessey, Lowry and Brock method Continuous flow analysis on Technicon SMA 12/60 Atomic absorption spectrophotometer Flame photometer Ion selective electrode
   Biospecimen Aliquots and Components	Hemolysis	Chemically-induced Hemolysate added No hemolysate added

   View more