Clinical Chemistry
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Clinical Chemistry 44: 2277-2280, 1998;
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(Clinical Chemistry. 1998;44:2277-2280.)
© 1998 American Association for Clinical Chemistry, Inc.

Enzymes and Protein Markers

HPLC detection of fetal blood in meconium: improved sensitivity compared with qualitative methods

Dan Chen1, Timothy R. Wilhite2, Carl H. Smith1,2, Morey A. Blinder1, and Michael Landt1,2,a

Departments of
1 Pathology and
2 Pediatrics, Washington University School of Medicine, St. Louis, MO 63110.
a Address correspondence to this author at: Department of Pediatrics, Washington University School of Medicine, One Children's Place, St. Louis, MO 63110. Fax 314-454-2274; e-mail landt{at}kids.wustl.edu.

We describe an HPLC-based method for the detection and quantification of fetal hemoglobin in stools of newborns. The new procedure is an alternative to the classic qualitative test for adult hemoglobin in meconium based on the differential stability of hemoglobin species in dilute base (Apt test). The HPLC method, based on a commercial device for hemoglobin characterization (Bio-Rad Variant), readily separates fetal and adult hemoglobin from non-hemoglobin components of meconium. To validate the method, blood and meconium were mixed in various proportions and then prepared for analysis with extraction in saline. The HPLC method accurately identified hemoglobin species even when the blood constituted only 5 mL per 100 g of the meconium specimen, and nearly quantitative recovery of hemoglobin was obtained at a blood content of 20 mL per 100 g of the meconium. Analysis time was 6.5 min, and preparation of sample was simple. HPLC detection of fetal blood in stools or other specimens markedly improves detection/characterization of blood in meconium.






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Copyright © 1998 by the American Association for Clinical Chemistry.