Influence of acid hydrolysis, saponification and sample clean‐up on the measurement of phytosterols in dairy cattle feed using GC–MS and GC with flame ionization detection

The fortification of processed foods including dairy products is increasingly commonplace with phytosterols among many compounds used to improve the nutritional value of food products. It is also increasingly common practice for some dairy cattle feeds to be fortified for their potential to increase phytosterol levels in milk. In this study, a combined, streamlined protocol using acid hydrolysis, saponification and sample clean-up was developed to enable the rapid and reliable measurement of phytosterols. The method was developed with focus on streamlining the overall technique to make it suitable for commercial laboratories, to reduce labor and consumable costs, while maintaining accuracy. A total of 12 different feed types commonly used in the dairy industry were analyzed with the highest and lowest sterol contents found in cotton seed oil and tannin with average phytosterol contents of 256 and <30 mg per 100 g, respectively. With a limit of reporting of 30 mg/kg for individual sterols and a correlation coefficient > 0.99, the method was validated for milk to enable feed comparison studies with respect to the total phytosterol content in raw milk.