Method Validation for Simultaneous Determination of Cholesterol and Cholesterol Oxides in Milk by RP-HPLC-DAD

Bauer, Luciana Carolina; Santana, Débora de Andrade; Macedo, Marianne dos S.; Torres, Alexandre Guedes; Souza, Nilson Evelázio de; Simionato, Julliana Izabelle

doi:10.5935/0103-5053.20130283

Cited by 24 publications

(28 citation statements)

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“…The most appropriate and frequently used steps for the sample preparation before liquid chromatography include the direct saponification followed by the extraction of the unsaponifiable residue into the nonpolar solvent [ 14 ]. Direct saponification has been preferred due to possibility to convert nonpolar fatty acid esters to polar products with their following effective removal by multiple extraction with n-hexane [ 14 , 15 , 16 , 17 ]. The other options can be single extraction by toluene [ 13 ], or three-stage extraction with diethyl ether [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials

Kolarič

Šimko

2020

Foods

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This work deals with up-to-date optimization of cholesterol content determination when saponification and extraction procedures as well as HPLC conditions were studied. As found, optimal conditions for saponification process were identified by 15 min heating in the presence of 0.015 L of methanolic KOH solution with a concentration 1 mol/L with subsequent 0.015 L n-hexane–chloroform binary mixture (1:1, v/v) double extraction. HPLC separation consisted of isocratic elution with flow rate of 0.5 mL/min mobile phase composed of acetonitrile/methanol 60:40 (v/v) and stationary phase Zorbax Eclipse Plus C18 column 2.1 × 100 mm, 3.5 μm particle size diameters with detector wavelength 205 nm. The method passed through in-house validation criteria and its suitability was verified by analysis of butter reference materials. In final, the average content of cholesterol content in butter was determined at 2271.0 mg/kg. Thus, the method is suitable for the determination of cholesterol content in butter and probably also in other dairy products.

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Section: Introductionmentioning

confidence: 99%

Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials

Kolarič

Šimko

2020

Foods

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“…Milk contains approximately 12 mg cholesterol/100 g (Walte, 1994), however, the cholesterol content in milk samples may vary C 2018 Institute of Food Technologists ® Vol. 17, 2018 r Comprehensive Reviews in Food Science and Food Safety 1475 due to the influence of various factors, such as: milk processing, differences in the breeds, and individual characteristics of the animals, milking intervals, lactation phase, composition of feed, as well as seasonal and climatic interferences (Bauer et al, 2014).…”

Section: Milk and Dairy Productsmentioning

confidence: 99%

Use of Natural Antioxidants in the Inhibition of Cholesterol Oxidation: A Review

Oliveira

Ferreira

Cople

et al. 2018

Comp Rev Food Sci Food Safe

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Synthetic antioxidants are widely used in the food industry. However, the potential toxicity, carcinogenic effects, and possible health damage caused by the ingestion of synthetic compounds, and also consumer concern about the safety of such additives has motivated the food industry to search for natural alternatives. Natural compounds with antioxidant properties are able to retard or prevent lipid oxidation in food. Animal sources like fish, eggs, meats, and dairy products are essential foods for human health due to their lipid fraction with high contents of unsaturated compounds, such as polyunsaturated fatty acids and cholesterol. However, these unsaturated lipids when exposed to favorable factors can become oxidized, which leads to sensory and nutritional losses as well as the formation of oxidized compounds known as cholesterol oxidation products or COPs. COPs are associated with deleterious health effects, such as inflammation, cytotoxicity, atherogenesis, carcinogenesis, and alterations in cell membrane properties, as well as the development of degenerative diseases such as Alzheimer's, Parkinson's, Huntington's, and other chronic diseases. Thus, the use of natural antioxidants can be an alternative to synthetics to prevent the formation of COPs and extend the shelf life of foods susceptible to oxidative deterioration. This review brings together information concerning the use of natural antioxidants as a strategy to control cholesterol oxidation.

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“…In contrast, the latter consist of less preparation steps resulting in faster analysis and a lower probability of artifact generation [23]. HPLC separations of the target analytes from various matrixes, using normal as well as RP chromatography combined with ultra violet (UV) [24][25][26][27], refractive index (RI) [28] or MS detection [29][30][31][32][33][34][35][36] have been previously reported. Application of MS detection not only allows the detection of all COPs, including those that completely lack chromophores like 5,6-epoxy-cholesterol, but is also more sensitive in comparison to the optical detection methods, reaching detection limits for some COPs as low as 10 ng/mL [31].…”

Section: Introductionmentioning

confidence: 99%

Quantification of sterols from carp cell lines by using HPLC–MS

Beck

Jordan

Herlitze

et al. 2018

Separation Science Plus

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Cholesterol is a major constituent of cell membranes and exhibits together with its oxidation products (oxysterols) several essential cellular functions. Additionally, it was recently shown to play a role during virus replication in fish. To investigate the involvement of cholesterol and oxysterols, for example, during a viral infection in fish cells, quantitative analysis of these analytes from cell cultures is necessary. A fast method based on solvent extraction followed by reversed‐phase high‐performance liquid chromatography with mass spectrometry for quantitation of cholesterol and seven oxysterols from cells and culture medium is described in this work. Detection limits were in the low ng/mL range, and the intra‐ and inter‐day precisions were above 87% for all analytes. A cholesterol content of approximately 10 μg per 2 × 106 cells, exceeding the concentration of oxysterols by at least 103‐fold, was determined for common carp brain cells. Furthermore, using the method established here, an uptake of externally supplied 25‐hydroxycholesterol by fish cells and its conversion to 7a,25‐dihydroxycholesterol could be shown. In summary, this is the first report on quantification of cholesterol and oxysterols from fish cell cultures, which can help in exploring their function during viral infections.

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Method Validation for Simultaneous Determination of Cholesterol and Cholesterol Oxides in Milk by RP-HPLC-DAD

Cited by 24 publications

References 0 publications

Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials

Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials

Use of Natural Antioxidants in the Inhibition of Cholesterol Oxidation: A Review

Quantification of sterols from carp cell lines by using HPLC–MS

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