HPLC DETECTION OF FETAL BLOOD IN MECONIUM - IMPROVED SENSITIVITY COMPARED WITH QUALITATIVE METHODS

We describe an HPLC-based method for the detection and quantification of fetal hemoglobin in stools of newborns. The new procedure is an alt ernative to the classic qualitative test for adult hemoglobin in mecon ium based on the differential stability of hemoglobin species in dilut e base (Apt test). The HPLC method, based on a commercial device for h emoglobin characterization (Bio-Rad Variant), readily separates fetal and adult hemoglobin from non-hemoglobin components of meconium. To va lidate the method, blood and meconium were mixed in various proportion s and then prepared for analysis with extraction in saline. The HPLC m ethod accurately identified hemoglobin species even when the blood con stituted only 5 mt per 100 g of the meconium specimen, and nearly quan titative recovery of hemoglobin was obtained at a blood content of 20 mt per 100 g of the meconium. Analysis time was 6.5 min, and preparati on of sample was simple. HPLC detection of fetal blood in stools or ot her specimens markedly improves detection/characterization of blood in meconium.