Stabilization of blood homocysteine in an epidemiological setting

The major problem in the determination of homocysteine (Hcy), which is thought to be a risk factor in colorectal cancer, is the rise in its concentration if blood is not centrifuged immediately after collection. We assess the interference of 3-deazaadenosine (which inhibits conversion of S-adenosylhomocysteine into Hcy within the erythrocyte), using the fluorescence polarization immunoassay (FPIA) assay, the stabilizing effect of 3-deazaadenosine and the impact of temperature on Hcy stabilization. To assess interference of 3-deazaadenosine, 12 blood samples were extracted; two aliquots were obtained from each and one of them was added 3-deazaadenosine (50 micromol/l). To assess the stabilizing value of 3-deazaadenosine, as well as the effect of temperature, two blood samples were extracted from 24 volunteers. One of the tubes was immediately placed on ice and centrifuged (reference concentration). To the second tube was immediately added 3-deazaadenosine (50 micromol/l), producing six aliquots, three of which were kept at room temperature (25 degrees C) for 1, 4 and 6 hours, the other three kept at 37 degrees C. The mean values (standard deviation) obtained for methodological interference were: 7.32 (3.58) micromol/l without stabilizer, and 7.11 (3.61) micromol/l with stabilizer. There were no statistically significant differences (P = 0.104) and intraclass correlation coefficient was 0.989, suggesting no methodological interference. We did not find any significant differences regarding our reference value in the samples kept at room temperature during the interval studied. A high Pearson correlation coefficient was obtained. Nevertheless, in those samples kept at 37 degrees C, a slight increase was observed in the 4-hour period (P = 0.009). The addition of 3-deazaadenosine may avoid problems in the critical pre-analytical phase in the Hcy measurement. There is no interference with the FPIA assay, nor any dilution effect, and new reference values are not necessary.