USE OF COMMERCIALLY AVAILABLE HEMOGLOBIN STANDARDS TO QUANTITATIVELY CALIBRATE A HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY METHOD

High performance liquid chromatography (HPLC) has proven to be an extr emely useful analytical technique to separate and identify different t ypes of hemoglobins, particularly A, C, F and S in blood samples, and compute their relative percentages. Such data provide useful informati on in the diagnosis of hemoglobinopathies including sickle cell anemia , P-thalassemia, hemoglobin C disease,etc. In the present investigatio n, we have explored the determination of absolute concentrations of in dividual hemoglobins in g/dL and recommend it as an additional paramet er which could be included as part of clinical data. Possible correlat ions between g/dL hemoglobin and the severity of a specific hemoglobin opathy could be established and aid in the diagnosis and/or treatment of such diseases. Several commercially available hemoglobin standards were analyzed and evaluated for use in creating g/dL calibration graph s for the HPLC. Appropriate calibration procedures have been developed and are presented. Also, linear regression graphs and sample chromato grams are included. Preliminary results obtained demonstrate the feasi bility of using commercially available hemoglobin standards to calibra te an HPLC method for the estimation of absolute hemoglobin concentrat ions.