7-Ketocholesterol and 7-ketositosterol were chosen as reliable markers of the oxidation of cholesterol and phytosterols in infant milk formulas and infant milk cereals. A reversed-phase HPLC method was developed to measure them simultaneously in infant formulas. This method was then tested on a wide range of infant milk formulas and milk cereals on sale in Italy whose lipid composition is representative of the most common commercial formulas. The analytical results revealed no significant differences in the extent of oxidation of cholesterol and sitosterol. As the level of 7-ketocholesterol often followed the cholesterol level, a cholesterol content similar to that of human milk produced amounts of cholesterol oxides with possible negative effects on infant health. In contrast, the low cholesterol content of milk cereals never produced amounts of cholesterol oxides high enough to cause concern. The contents of phytosterols and hence their oxides were always low.

The development of the Maillard reaction in pasteurized, UHT and in-bottle sterilized dietetic milks was studied. In these products damage caused by heat treatments could increase as a result either of the addition of various ingredients or of manufacturing processes that alter their content of reducing carbohydrates. Protein damage was evaluated by measuring furosine by reversed-phase ion-paired HPLC. The levels of furosine detected made it possible to assess the amounts of biologically unavailable lysine. In all milks analysed blocked lysine values were <340–350 mg/g total lysine, the level at which lysine becomes the limiting amino acid in milk. Pasteurized dietetic milks had levels of blocked lysine similar to that in ordinary pasteurized cows' milk. In some UHT and in-bottle sterilized dietetic milks their different composition resulted in an increase in the blocked lysine content. In some in-bottle sterilized milks, protein damage greatly reduces the beneficial effects of milk as a dietary supplement. Lactose-free milks, which are more susceptible to protein deterioration because of their higher content of reducing carbohydrates, were also analysed after storage at 20°C and at [les ]4°C. At the end of their recommended storage times, they contained limited amounts of blocked lysine only if they had been stored at [les ]4°C.

The quantity of blocked lysine in spray-dried infant formulas in Italy was calculated by the furosine method. This quantity was then correlated with the composition of the carbohydrate fraction. In formulas containing only lactose as carbohydrate its presence in large quantities (6.7-7.5 g/l00 mL of formula as fed), with a lysine-rich protein fraction, may lead to amino acid blocking of >20%. In formulas in which lactose was substituted with low-DE maltodextrin a reduction of lysine blocking was possible. Addition of glucose may result in high levels of blocked lysine.

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Cholesterol oxidation in commercial meat-based homogenized and freeze-dried baby foods was examined. The 7 major products of this reaction were determined by gas chromatography coupled with mass spectrometry (GC-MS). As far as single cholesterol oxidation products (COP) are concerned, 7-ketocholesterol was the major product of direct cholesterol oxidation in the 2 groups of analyzed samples, and this study confirmed that it is a useful marker of the whole cholesterol oxidation process. Nevertheless, the amounts of cholesterol-5β,6β-epoxide were often similar to and sometimes higher than the amounts of 7-ketocholesterol, thus showing a strong development of both direct and indirect cholesterol oxidation pathways. Total COP content was significantly higher in freeze-dried than in homogenized products. Moreover, in freeze-dried samples, the COP content per serving was quite variable and, in 2 samples, it was close to or even higher than 500 μg. The greater development of cholesterol oxidation in the freeze-dried samples was confirmed by their highest total COP/cholesterol percent ratios. A constant correlation between the fatty acid composition and the development of cholesterol oxidation was not found, although a positive correlation between unsaturated fatty acid content and total COP content occasionally exists in samples of the same brand.