Cholesterol Content in Human Milk during Lactation: A Comparative Study of Enzymatic and Chromatographic Methods

This study validates a gas chromatography (GC) method for determining the sterol profile of human milk (HM) and compares it with an enzymatic-spectrophotometric (E-S) method. Good linearity ( r > 0.97) and low limits of detection and quantification were obtained with the GC method (<1.8 and <6 μg/100 g of HM, respectively). Suitable intra- and interassay precisions (all <18%) and satisfactory recovery percentages (80-109%) were obtained for both methods. In addition, both methodologies were used to assess cholesterol evolution in HM during lactation, showing a 50% decrease at 6 months versus colostrum. The E-S method overestimated cholesterol content by <20% versus the GC method. The results indicate that both methods may be used by the industry and in research to better understand the differences between the sterol profiles of infant formulas and HM.