Primus Corporation

Ching-Nan Ou

Jimmie K Noffsinger

Improved HPLC methods for the identification and characterization of hemoglobin variants in blood samples are disclosed. These methods are performed manually or with the aid of a computer. In preferred embodiments, a blood sample is injected into an HPLC unit and hemoglobin species from the blood sample are separated using high column temperatures and high eluant flow rates. Blood samples are initially screened using a low-resolution HPLC method to determine if an eluted hemoglobin species has a characteristic indicative of a hemoglobin variant, i.e., if a chromatograph peak of the eluted species has one of a number of parameter values falling within a predetermined range of values. If the initial screening indicates that a blood sample may have a hemoglobin variant, the blood sample is subsequently reanalyzed using a high-resolution HPLC method to determine if an eluted hemoglobin species corresponds to a known hemoglobin variant by comparing the retention time of an eluted species to the retention times of known hemoglobin variants. Alternatively, both the low-resolution and high-resolution HPLC methods can be used independently of one another. Thus, the methods of the invention are useful in putatively identifying abnormal hemoglobins.

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Ching-Nan Ou