Comparative of detection method of free hemoglobin in ventricular assist device hemolysis test
Objective To evaluate three methods for detecting free hemoglobin in hemolysis tests and determine the most suitable method for ventricular assist devices.Methods By conducting a comparative study,we describe the principles and methods of the Benzidine,Harboe,and Cripps methods for hemoglobin content testing.Standard hemoglobin samples of different concentrations were tested to plot standard curves depicting the relationship between hemoglobin concentration and absorbance.The advantages and disadvantages of the three methods were analyzed regarding experimental operation and result variability.Additionally,the impact of 1％ and 0.2％ ortho-tolidine solutions on the Benzidine method and the effect of different diluent concentrations (0.01％ sodium carbonate solution and 0.1％ sodium carbonate solution) in the Harboe method were compared.Results The Cripps method showed poor repeatability with a low fitting coefficient (R2=0.8019) and failed to produce a high-quality standard curve.The Benzidine and Harboe methods exhibited good repeatability and could yield high-fitting standard curves (both with R2>0.99) .The Benzidine method is recommended to use a 1％ ortho-tolidine solution,and the Harboe method is more suitable with a 0.01％ sodium carbonate solution.However,the Benzidine method requires strict reagent preparation and immediate use,with time-dependent measurement results.Conclusions Compared to other testing methods,the Harboe method can overcome the shortcomings of the other two methods.It is suitable for rapid automated analysis in laboratories and is more appropriate for hemolysis testing in ventricular assist devices.
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