HPLC separation and determination of 12 cholesterol oxidation products in fish: comparative study of RI, UV, and APCI-MS detectors

A simple, fast, and sensitive method for the extraction through direct saponification, separation, quantification, and identification of 12 cholesterol oxidation products (COPs) and cholesterol in a single isocratic, normal-phase, high-performance liquid chromatography (HPLC) was developed. Three detectors were compared for determination of COPs and cholesterol in fish samples: refractive index (RI), ultraviolet (UV), and atmospheric pressure chemical ionization mass spectrometry (APCI-MS). The results did not show significant differences (p > 0.05) between the concentration of the cholesterol oxides and cholesterol obtained with these detectors. The present study demonstrated the presence of 19-hydroxycholesterol, 22"R"-hydroxycholesterol, 22"S"-hydroxycholesterol, 24"S"-hydroxycholesterol, and 25"R"-hydroxycholesterol for the first time in fish samples.     

Cholesterol oxides,cholesterol, total lipid,and fatty acid composition in turkey meat

The contents of cholesterol oxides, cholesterol, and total lipid, and the fatty acid composition were determined in frozen turkey meat. The 7-ketocholesterol content varied from 33 microg/100 g in the breast to 765 microg/100 g in the skin, and the levels of 7 beta-hydroxycholesterol varied from not detected in the leg, breast, and skin to 370 microg/100 g in the skin. The values for total lipid (g/100 g) in the wings, legs, breast, and skin were 0.9 +/- 0.4, 1.1 +/- 0.2, 0.5 +/- 0.1, and 12 +/- 3, respectively. The contents for cholesterol (mg/100 g) were 46 +/- 5, 35 +/- 2, 27 +/- 3, and 81 +/- 6 in the wing, legs, breast, and skin, respectively. The main fatty acids identified in all cuts were C18:2n6, C18:1n9, C16:0, C18:0, and C20:4n6.     

Effect of storage on cholesterol oxide formation and fatty acid alterations in egg powder

The effect of the storage of egg powder on cholesterol oxide formation and alterations in the fatty acid composition was studied. Two commercial brands, A and B, were studied at 0 time and then monthly for up to 6 and 12 months, respectively. Five cholesterol oxides were identified and quantified (7-ketocholesterol, 7beta-hydroxycholesterol, 7alpha-hydroxycholesterol, 5,6alpha-epoxycholesterol, and 5,6beta-epoxycholesterol), the amounts of which increased during storage, although the cholesterol contents remained constant. The polyunsaturated fatty acid contents reduced during storage, whereas the levels of saturated and monounsaturated fatty acids and total lipids remained constant. High cholesterol oxide and trans fatty acid contents were found, a fact of concern because these compounds are hazardous to health, and egg powder is used in various food products widely consumed by the population, principally by infants.     

Evaluation of the ratio of omega(6: omega3 fatty acids and vitamin E levels in the diet on the reproductive performance of cockerels.

Three hundred and twenty 30-week old White Leghorn cockerels were housed in individual cages and distributed in a completely randomized factorial design of 5 x 3, with five oil sources (sunflower, soybean, canola, linseed and fish/soybean) and three levels of antioxidant (30, 200 and 400 mg of vitamin E/kg). The aim of this study was to evaluate the effect of the ratio of omega6: omega3 fatty acids by the inclusion of different oil sources and of dietary supplementation with vitamin E on the reproductive performance of cockerels. The use of the fish/soybean combination determined the lowest total antioxidant status of the semen. However, the addition of vitamin E to the fish/soybean-oil-based diet resulted in a linear increase in semen volume, motility and sperm vigour in the 38th week and again in the 52nd week for motility and for sperm vigour and fertility rate in the periods from 50-53 and 41-53 weeks of age. The use of canola oil in the diet resulted in the highest fertility rate during 50-53 and 41-53 weeks of life. Animals receiving the soybean oil based diet showed the smallest fertility rate in the range from 50-53 weeks of age and concomitantly the highest level of cholesterol in the spermatozoa in the range from 47-51 weeks. An interaction between the vitamin E level and soybean oil was verified by the linear increase in motility and sperm vigour at 38 weeks of age. Later, the contrary was shown by a linear reduction in fertility in the periods from 44-46, 47-49 and 41-53 weeks of age. Cockerels that had been fed on the sunflower-oil-based diet showed the highest content of saturated fatty acids in the spermatozoa from 48-51 weeks. An interaction effect was observed between the vitamin E level and sunflower oil shown by a linear increase in the content of saturated fatty acids in the spermatozoa and a linear reduction in the C18: 1omega9, C18 :2 omega6 and PUFA (C18 : 2omega6 + C20 : 4omega6) contents in the spermatozoa at 48-51 weeks and in sperm volume at 52 weeks of age.     

Optimization and validation of analytical conditions for cholesterol and cholesterol oxides extraction in chicken meat using response surface methodology

The analytical conditions for the extraction of cholesterol and cholesterol oxides in chicken meat were optimized by means of response surface methodology. The separation and identification were performed by normal phase HPLC using UV and refractive index (RI) detectors, and the confirmation of the 11 cholesterol oxides identities in the samples was verified by HPLC-APCI-MS. The developed methodology showed good analytical performance, presenting recovery levels from 84 to 103% and detection limits varying from 0.01 to 0.06 microg/g for UV detection and from 1.98 to 2.12 microg/g for RI detection. The present study demonstrated the presence of 22 R-hydroxycholesterol, 24 S-hydroxycholesterol, and 22 S-hydroxycholesterol for the first time in chicken meat.