Development of a high-pressure liquid chromatography method for diagnosis of heparin-induced thrombocytopenia.
Owing to the disadvantage of radioactivity of the carbon 14 serotonin release assay and the time-consuming procedure of the enzyme immunoassay, we developed a high-pressure liquid chromatography (HPLC) method to detect serotonin released from donor platelets in the presence of heparins and serum samples from patients with heparin-induced thrombocytopenia (HIT). Samples were analyzed from 60 healthy control subjects, 19 patients with HIT, and 20 patients without HIT after incubation with heparin, low-molecular-weight heparin (LMWH), and danaparoid. Serotonin release was measured from platelets, 300 x 10(3)/microL, by HPLC. Serotonin eluted as a single peak from the HPLC column. Serum samples from patients with HIT released 5.5 to 352.5 and 6.6 to 1,533.3 ng/mL of serotonin from platelets in the presence of 0.2 IU/mL of heparin and LMWH, respectively. In the presence of 0 IU/mL of heparin, LMWH, danaparoid, and control samples, less than 2.5 ng/mL of serotonin were released. The HPLC method permits a rapid, sensitive, and quantitative determination of serotonin released from donor platelets for laboratory confirmation of HIT.
Koch
Sandra
S
IV Department of Medicine, University Hospital, Faculty of Clinical Medicine Mannheim of the Ruprecht-Karls-University of Heidelberg, Heidelberg, Germany.
Harenberg
Job
J
Odel
Michael
M
Schmidt-Gayk
Heinrich
H
Walch
Stefan
S
Budde
Uwe
U
D000925:Anticoagulants / Q000009:adverse effects*; D001792:Blood Platelets / Q000378:metabolism; D002809:Chondroitin Sulfates / Q000009:adverse effects; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003871:Dermatan Sulfate / Q000009:adverse effects; D004338:Drug Combinations; D006495:Heparin, Low-Molecular-Weight / Q000009:adverse effects*; D006497:Heparan Sulfate / Q000009:adverse effects; D006801:Humans; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D012701:Serotonin / Q000378:metabolism; D016037:Single-Blind Method; D013921:Thrombocytopenia / Q000139:chemically induced* / Q000175:diagnosis* / Q000378:metabolism
D000925:Anticoagulants; D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D004338:Drug Combinations; D006495:Heparin, Low-Molecular-Weight; D006497:Heparan Sulfate; D012701:Serotonin; C035838:danaparoid