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Method for Recovery of Progesterone Added to Milk¹

Department of Dairy Science and Department of Food Science and Biochemistry, Clemson University, Clemson, South Carolina 29631

ABSTRACT

A relatively simple technique for extraction of progesterone from milk was developed. Approximately 0.4 µg (3,500 count/min) of progesterone-4-¹⁴C was added to each milk sample before extraction. The milk sample was subjected to total lipid extraction and two solvent partitioning procedures, with the final residue redissolved in 95% ethanol. Scintillation counting of aliquots of the final extract indicated a recovery of 72.9 ± 3.8% (mean ± SD) of the added hormone. The major portion (97.3 ± 1.6%) (mean ± SD) of the loss of radioactivity occurred during the two solvent partitioning phases. The total recovery of isotope, including both the experimental extract and waste products, was 98.7 ± 4.8% (mean ± SD). The recovery rate for this method compares well with methods reported for hormone assay of animal tissue. Spectrophotometric determinations of authentic progesterone added to milk confirmed that the recovered radioactive substance was progesterone. The method is efficient enough to extract small quantities of progesterone for investigating progestins in milk.

¹ Technical contribution number 776, South Carolina Agricultural Experiment Station, Clemson, South Carolina. Published with the approval of the Director.