Effect of irradiation and packaging conditions after cooking on the formation of cholesterol and lipid oxidation products in meats during storage

The effect of irradiation and packaging conditions on the content of cholesterol oxidation products (COPs) and lipid oxidation in cooked turkey, beef, and pork during storage was studied. Ground turkey leg, beef, and pork were cooked, packaged either in oxygen-permeable or oxygen-impermeable bags, and irradiated at 0 or 4.5 kGy. Lipid oxidation and COPs were determined after 0 and 7 days of storage at 4°C. Packaging of cooked meat was more important than irradiation in developing COPs and lipid oxidation in cooked meats during storage. 7-Hydroxycholesterol, 7β-hydroxycholesterol, β-epoxide, and 7-ketocholesterol were among the major COPs formed in cooked turkey, beef, and pork after storage, and their amounts increased dramatically during the 7-day storage in aerobic conditions. Irradiation had no significant effect on the amounts of any of the COPs found in cooked turkey and beef, but increased (P<0.05) the amounts of - plus 7β-hydroxycholesterol, β-epoxide, 7-ketocholesterol, and total COPs in aerobically packaged cooked pork. The amounts of COPs and lipid oxidation products (TBARS) closely related to the proportion of polyunsaturated fatty acids in meat. The results indicated that the composition of fats in meat is important on the oxidation rates of lipids and cholesterol, and packaging is far more important than irradiation in the formation of COPs and lipid oxidation in cooked meat.     

Evaluation of cholesterol and lipid oxidation in raw and cooked minced beef stored under oxygen-enriched atmosphere

Oxygen-enriched modified atmosphere packaging (MAP) represents an important means to stabilize meat colour but may lead to an increase in lipid oxidation, influencing the acceptability and safety of the product. In this work, the effect on cholesterol and lipid susceptibility to oxidation was investigated in commercial minced beef held under MAP (80% O(2)/20% CO(2)). Cholesterol oxidation products (COPs), peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) were determined, before and after pan frying, at 1, 8 and 15 days since packaging under refrigerated storage (3-4°C). 7α-Hydroxycholesterol, 7β-hydroxycholesterol and 7-ketocholesterol were the more abundant COPs identified. COPs significantly increased in raw beef during storage: after 1, 8 and 15 days since packaging COPs were at the levels of 10.4, 30.7 and 60.5μg/g of fat, respectively. Cooking did not affect cholesterol oxidation in freshly packaged minced beef but led to a rise in COPs amount with respect to raw muscle after 8 and 15 days of storage. The trend in cholesterol oxidation reflected the progressive increase in lipid peroxidation rate brought by MAP conditions.     

Quantification of Three Cholesterol Oxidation Products in Raw Meat and Chicken

Raw veal, beef, pork, and chicken muscle tissues were extracted by a modified dry column procedure in which silicic acid was incorporated in the trap of the column. This method separated cholesterol derivatives from the bulk of neutral lipids, phospholipids and cholesterol. Isolation of sterols by preparative thin layer chromatography followed by quantification by direct on-column capillary gas chromatography permitted measurement of 7-ketocholesterol, cholesterol 5α, 6α-epoxide, and cholesterol 5β 6β-epoxide at concentrations less than 1 ppm. All muscle tissues contained the three cholesterol products in measurable quantities. 7-Ketocholesterol constituted more than 50% of the oxidation products in all samples.     

Cholesterol oxidation products in irradiated raw meat with different packaging and storage time

The effect of irradiation and packaging conditions on the formation of cholesterol oxidation products (COPs) as well as lipid oxidation products was determined in raw turkey leg, beef, and pork loin meat during 7 days of storage. Ground turkey leg, beef, and pork loin muscles were prepared as patties. The patties were individually packaged either in oxygen-permeable or impermeable bags, irradiated at 0 or 4.5 kGy using a Linear Accelerator, and stored at 4°C. The COPs such as 7-hydroxycholesterol, 7β-hydroxycholesterol, and 7-ketocholesterol were detected in fresh raw meats at 0 day at the level of 10.9 to 49.2 μg/g lipid. After 7 days of storage, other COPs such as epoxides, 20-hyroxycholesterol, and choletanetriol were formed in mainly aerobically packaged and irradiated raw meats. Packaging effect was more crucial on the cholesterol and lipid oxidation than irradiation. In aerobically packaged and irradiated meats, turkey leg muscles had higher COPs value than beef or pork did. COPs and thiobarbituric acid reactive substances (TBARS) values had a strongly positive correlation in turkey leg and pork. But, cholesterol oxidation in beef proceeded in irradiated and aerobically stored samples despite of its low level of TBARS value.     

TBA Values and 7-Ketocholesterol in Refrigerated Raw and Cooked Ground Beef

Thiobarbituric acid (TBA) values and 7-ketocholesterol were determined on raw and cooked ground beef patties after 0, 2, and 4 days of storage at 4°C. The mean 7-ketocholesterol at days 0, 2, and 4 was 9.65, 23.8, and 42.3 μg for 100g raw patties and 6.33, 277.9, and 484.7 kg for 100g cooked patties. The TBA values increased after 2 and 4 days for the raw and cooked patties. TBA values and 7-ketocholesterol were correlated with each other for raw (r²= 0.82) and cooked (r²= 0.98) patties. The peak suspected to be 7-ketocholesterol was confirmed by mass spectral analysis. The 7-ketocholesterol in the cooked patties stored two days represented a 0.3% oxidation of the free cholesterol.     

Wide-Bore Capillary Gas Chromatographic Method for Quantification of Cholesterol Oxidation Products in Egg Yolk Powder

A wide-bore capillary gas chromatographic (GC) method was developed for quantification of cholesterol oxidation products in egg yolk powder. Baseline separations were achieved between 7α- and 7β-hydroxycholesterol, cholesterol, cholesterol-5α,6α-epoxide, cholesterol-5β,6β-epoxide and 7-ketocholesterol chromatographed as the trimethylsilylated sterols. Excellent response linearity was obtained for each cholesterol oxidation product, permitting reliable quantification. Recoveries from freeze-dried egg yolk spiked with various levels of cholesterol oxidation products ranged from 78.2 to 95.1%. The coefficients of variation compared favorably to those for packed column GC methods.     

Cholesterol Oxidation in Baked Foods Containing Fresh and Powdered Eggs

The degree of cholesterol oxidation in commercial sweet baked foods (biscuits and snacks) and in laboratory baked biscuits, all containing fresh or powdered eggs, was determined. 7-Ketocholesterol was used as index of cholesterol oxidation and detected by two analytical methods. The analysis of the biscuits showed higher levels of 7-ketocholesterol and a more marked oxidative instability of cholesterol when prepared with powdered eggs. The significant amounts of 7-ketocholesterol found in some samples of commercial biscuits were attributed to the use of powdered eggs. These data are of importance to industries using eggs in sweet baked products which are mainly consumed by children.     

Antioxidant Inhibition of Cholesterol Oxidation in a Spray-Dried Food System during Accelerated Storage

Spray-dried egg yolk was used to evaluate antioxidant inhibition of cholesterol oxidation during accelerated (CU⁺² and heat) storage. Liquid yolk was treated with equimolar amounts of BHA (0.01%, w/w of lipid), ascorbyl palmitate (0.023%), or a tocopherol blend (0.023%). Yolk batches were spray-dried, stored at 60 ± 2°C for up to 28 days, and analyzed for cholesterol oxidation products (COPS) using gas chroma-tography. COP levels generally increased during storage with 7-ketocholesterol predominant. Significant antioxidant effects were manifest in decreased levels of 7-ketocholesterol, 7α- and 7β-hydroxycholesterol; while cholestane-triol and cholesterol-5,6-epoxide levels were not affected. All antioxidants showed significant inhibitive effects relative to the control, and tocopherols were more effective than ascorbyl palmitate.     

Lipid and cholesterol oxidation in Chinese-style sausage using vacuum and modified atmosphere packaging

The oxidation of lipid and cholesterol in Chinese-style sausage in vacuum packaging (VP) and modified atmosphere packaging (MAP) stored at 4°C and 15°C, respectively, for 5 months was investigated. The 2-thiobarbitaric acid reactive substance (TBARS) and peroxide value (POV) of sausage were variable with packaging treatments during storage. TBARS and POV in sausage stored at 15°C were significantly greater (p < 0·05) than at 4°C, and the MAP treatment was more stable than the VP treatment. In addition, the polyunsaturated fatty acids (PUFA) in sausage decreased with storage for both treatments. The content of cholesterol decreased significantly after 3 months of storage. 7-β-hydroxycholesterol, 7-ketocholesterol and 22-ketocholesterol were the major cholesterol oxidation products (COPs), but there was no detectable (< 1 μg/100 g) 25-hydroxycholesterol or cholestanetriol with either treatment.     

Effect of dietary vitamin E supplementation on cholesterol oxidation in vacuum packaged cooked beef steaks

The effect of dietary vitamin E supplementation on cholesterol oxidation in vacuum packaged, cooked, refrigerated and frozen beef steaks, was investigated. Steers (Friesian×Charolais×Black Hereford) were fed diets providing 20 or 3000 mg α-tocopheryl acetate/head/day for 135 days prior to slaughter. α-Tocopherol concentrations in M. psoas major (PM) and M. longissimus dorsi (LD) were significantly (p<0.05) increased by supplementation and were significantly (p<0.05) higher in PM than LD. Cholesterol oxidation (monitored by measuring 7-ketocholesterol formation) increased during refrigerated and frozen storage in some, but not all, groups, and tended to be higher in PM than LD. Dietary vitamin E did not affect 7-ketocholesterol formation in LD, but significantly (p<0.05) reduced concentrations in PM during refrigerated and frozen storage. Supplementation significantly (p<0.05) reduced TBARS in PM and LD, indicating that vitamin E improved oxidative stability in both muscles. The results show that dietary vitamin E supplementation inhibits cholesterol oxidation in vacuum packaged, cooked beef during refrigerated and frozen storage, but may be influenced by muscle type.     

胆固醇热氧化反应研究

以纯胆固醇为对照，对油脂存在下胆固醇受热氧化情况进行了研究。探讨了温度、加热时间、花生油加入量对胆固醇氧化的影响。采用自制微型固相萃取柱样品预处理结合毛细管气相色谱法分离检测胆固醇氧化产物，共有6种氧化物被检出。在一定范围内，氧化物的组成和生成量随加热温度升高和时间延长明显增加，而随加油量的增加有明显减少的趋势。     

DIETARY OILS AND TOCOPHEROL SUPPLEMENTATION ON CHOLESTEROL OXIDE FORMATION IN FREEZE-DRIED CHICKEN MEAT DURING STORAGE

The influence of dietary oils [fish (FO), flax (LO), palm (PO) and sunflower (SO) oil] and tocohperol supplementation on the oxidation of cholesterol and lipids of chicken meat was investigated. Lipid oxidation in chicken meat powders was determined by thiobarbituric acid reactive substances (TBARS) and cholesterol oxidation products were estimated by capillary gas chromatography (GC) after trimethylsilylation. Peak identities from GC were confirmed by mass spectrometry. Freeze-dried chicken meat powder, stored in contact with air at room temperature for four months showed the presence of 7α- and 7β-hydroxycholesterol, 7-ketocholesterol, α and β-epoxide, and cholestane-triol. Total cholesterol oxides contents significantly (P < 0.05) increased with storage. The amount of cholesterol oxides in the meat powder was the highest in FO and SO group, and lowest in PO group. Lipid and cholesterol oxidation were accelerated by the presence of long-chain PUFA. Lipid and cholesterol stability of chicken meat were improved by the dietary supplementation of tocopherol.     

Influence of carcass weight on instrumental and sensory lamb meat quality in intensive production systems

The effects of cooking viz. pressure-cooking and broiling and storage at 4 °C for six days and -10 °C for 90 days on lipid oxidation and development of cholesterol oxidation products in mutton were studied. Results revealed that cooking of meat significantly increased the total lipids, phospholipids, cholesterol, glycolipids, free fatty acids and glycerides, but they did not change during refrigerated and frozen storage. The TBA values increased on cooking and during storage. However, the values were below the threshold level for rancidity development. The following cholesterol oxidation products were separated by thin layer chromatography cholestanetriol, 7-α-hydroxy cholesterol, 19-hydroxycholesterol, 7-ketocholesterol, cholesterol-α-epoxide, cholesterol-β-epoxide and an unidentified fraction. All these fractions except the unidentified fraction increased on cooking. On refrigerated and even on frozen storage all these fractions increased except the unidentified fraction, which showed a concomitant reduction. The changes in broiled meat were more pronounced compared to pressure-cooked meat. Results clearly indicated that even frozen storage of cooked meat did not prevent the development of cholesterol oxidation products.     

Effect of different cooking methods on some lipid and protein components of hamburgers

The effects of different cooking methods on the lipid and protein fractions of hamburger were evaluated. The lipid component was subjected to the following analyses: peroxide value; p-anisidine; total and free fatty acids; cholesterol and its oxidation products (quantified as 7-ketocholesterol). Lysinoalanine (LAL), free amino acids and D-amino acids (D-AA) were also determined in the protein fraction. All results were compared with a raw control. No significant differences were found among the cooking treatments with respect to D-AA and LAL. The degree of proteolysis, lipolysis and lipid oxidation varied depending on the treatment conditions. Regarding cholesterol oxidation, the combination of roasting and microwave heating caused more oxidation than the other treatments. The raw meat, however, showed an advanced degree of oxidation (25.2 ppm of total 7-ketocholesterol/120 g ground meat).     

THE EFFECTS OF DIETARY α-TOCOPHEROL AND SURFACE APPLICATION OF OLEORESIN ROSEMARY ON LIPID OXIDATION AND CHOLESTEROL OXIDE FORMATION IN COOKED RAINBOW TROUT (Oncorhynchus mykiss) MUSCLE

The inhibitory effect of dietary α-tocopherol supplementation (100 and 500 mg α-tocopheryl acetate/kg diet) and an oleoresin rosemary dip on lipid and cholesterol oxidation in cooked rainbow trout ( Oncorhynchus mykiss ) during storage at 4C for 48 h was investigated. The 2-thiobarbituric acid-reactive substances (TBARS) values of cooked fish muscle increased during storage for all treatments. Dietary α-tocopherol supplementation partially inhibited lipid oxidation. Surface application of oleoresin rosemary further enhanced this protective effect. Cholesterol oxides were formed in all the samples following cooking and storage. The major cholesterol oxidation products (COPS) were identified as 7 β-hydroxycholesterol, α- and β-epoxides, and 7-ketocholesterol. Formation of COPS was reduced by dietary supplementation with α-tocopherol and surface application of oleoresin rosemary. Statistical analysis revealed a highly significant (p < 0.01) inhibitory effect of oleoresin rosemary on COPS formation.     

Effect of cooking and storage on lipid oxidation and development of cholesterol oxidation products in water buffalo meat

Buffalo meat was subjected to two cooking methods viz. broiling and pressure cooking and two storage procedures viz. refrigerated (4 °C) storage for six days and frozen (-10 °C) storage for 90 days. Changes in lipid oxidation and development of cholesterol oxidation products were studied in raw as well as cooked meat samples. Total lipid, phospholipid, cholesterol, free fatty acid, glycolipid and glyceride contents increased significantly on cooking of meat but did not show any significant changes during either refrigerated or frozen storage except for free fatty acid content which showed an increase. The TBA values also increased during storage but not to the extent of indicating rancidity. Cholesterol oxidation products separated by thin layer chromatography were: cholestanetriol, 7-α-hydroxycholesterol, 19-hydroxycholesterol, 7-ketocholesterol, cholesterol-α-epoxide, cholesterol-β-epoxide and an unidentified fraction. All these fractions, except for the unidentified fraction, increased on cooking and storage. The cholesterol-β-epoxide fraction was resistant to changes. Changes in broiled meat were more pronounced compared to pressure cooked meat. Frozen storage did not prevent the development of cholesterol oxidation products in buffalo meat.     

Enzymatic Determination of Cholesterol Oxides

For measurement of the concentration of oxidised cholesterols in foodstuffs, a method was developed by adapting the well-known enzymatic determination of cholesterol. A linear correlation was found between the absorbance measured after enzymatic reaction and the concentrations of 7α-hydroxycholesterol, 7β-hydroxycholesterol, 7-ketocholesterol, cholesterol-5α, 6α-epoxide in the range 1–12 μg. The usefulness of this method was demonstrated by determination of the four cholesterol oxidation products separated by TLC from the non-saponifiable lipid fraction of 5-month-old whole-egg powder (stored under different conditions) and of biscuit enriched with egg powder. The main advantages of the combination of TLC and the enzymatic method, beyond its rapidity and sensitivity, are that the determination of cholesterol oxidation products can be carried out in parallel with many samples containing small amounts of oxidised cholesterols even in the presence of a large quantity of cholesterol.     

Cholesterol Autoxidation Inhibition Varies Among Several Natural Antioxidants in an Aqueous Model System

Inhibition of cholesterol autoxidation by several natural materials was examined in an aqueous meat model system at pH 5.50 and 80°C. Antioxidant effectiveness was measured using the induction period for 7-ketocholesterol. An industrial rosemary oleoresin, quercetin, myr-icetin and BHA (included for comparison) had no antioxidant properties for cholesterol. All tocopherol treatments delayed cholesterol oxidation. The γ- and δ- treatments were most effective and α-toco-pherol was least effective. No synergistic effects were observed with tocopherol blends. Antioxidants effective against unsaturated fatty acid autoxidation may not inhibit cholesterol autoxidation.     

Effect of Ionizing Radiation on Cholesterol in Aqueous Dispersion

Aqueous sodium stearate dispersions of cholesterol were irradiated at 0-2°C with absorbed doses ranging from 2.5 to 50 kGy. The resulting mixture of cholesterol derivatives was isolated and examined for 7-ketocholesterol and cholesterol 5α,6α-epoxide and 5β,6β-epoxide content. Concentrations of all three compounds increased with dose, while the ratio of 7-ketocholesterol to total epoxides decreased with increasing dose. The ratio of 7-ketocholesterol to the epoxides was approximately 1 or below at all dose levels while the same ratio in autoxidations of cholesterol in dispersions was normally 6 or greater. The change in the keto/epoxide ratio may be a means for determining whether meat or other foods containing cholesterol have been subjected to ionizing radiation.     

Influence of packaging atmosphere on the formation of cholesterol oxides in γ-irradiated egg powder

In a previous work the effect of ionising radiation on formation of cholesterol oxidation products (COPs) under aerobic conditions was studied. In the present work the influence of aerobic and anoxic conditions have been comparatively investigated to study the chemical changes of cholesterol in γ-irradiated egg powder. The irradiation treatment was carried out with powdered egg packed under air and also under vacuum in polyethylene (PE) bags and in laminated, oxygen impermeable three-layer (polyester-aluminium-polyethylene) foil to dosage levels 2 and 4 kGy at room temperature. The cholesterol oxidation is demonstrated by thin-layer chromatograms. In the egg powder wrapped in PE bags the predominant cholesterol derivatives?7-hydroxycholesterol isomers (α and β)—accumulated in significant amounts (increasing with dose) while vacuum-packaging in laminated foil considerably suppressed the quantity of these products and prevented the formation of cholesterol 5α, 6α-epoxide as well as 7-ketocholesterol. Little or no change was observed in non-irradiated (control) vacuum-packed egg powder stored at approximately 22°C for up to 5 months. Peroxide values showed changes parallel to the formation of COPs.