Total lipids were extracted from the mammary glands and milk obtained on LD10 according to the methodology of Folch et al. (1957) . The lipid extracts were fractionated into phospholipids and neutral lipid fractions by the micro-column chromatography method (Hurtado de Catalfo et al., 2009) and quantified gravimetrically (Marra and de Alaniz, 1990) . After the saponification of all milk samples at 80 °C for 45 min under a nitrogen atmosphere in 10% KOH in ethanol, the free FAs were extracted with hexane and evaporated to dryness. Then, they were esterified (de Alaniz and Marra, 1992) , and the FA methyl esters (FAMEs) obtained were extracted twice with hexane. FAMEs were quantified by gas chromatography using a capillary-GLC-Hewlett Packard HP 6890 GC Series System Plus (Avondale, PA, USA) and were identified by comparisons of their relative retention times with authentic standards. The oven temperature was programmed to increase from 185 to 230 °C in increments of 3 °C/min. Ultra-dry helium was used as a carrier gas at an operating pressure of 3.0 kg/cm 2 . FAMEs were detected with a flame ionization detector operating at 280 °C. In some cases, the identities of the FAs were confirmed by mass spectrometry analysis. Each FA was expressed as μmol per mg of lipids in the sample.
Hp 6890 gc series system plus
Manufactured by Hewlett-Packard
Sourced in United States
The HP 6890 GC Series System Plus is a gas chromatography system designed for analytical and research applications. It features a programmable oven, multiple injection ports, and advanced data processing capabilities. The system is capable of separating, identifying, and quantifying components in complex mixtures.
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2 protocols using hp 6890 gc series system plus
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Quantitative Lipid Analysis in Mammary Tissue
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Quantitative Lipid Analysis in Mammary Tissue
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Total lipids were extracted from the mammary glands and milk obtained on LD10 according to the methodology of Folch et al. (1957) . The lipid extracts were fractionated into phospholipids and neutral lipid fractions by the micro-column chromatography method (Hurtado de Catalfo et al., 2009) and quantified gravimetrically (Marra and de Alaniz, 1990) . After the saponification of all milk samples at 80 °C for 45 min under a nitrogen atmosphere in 10% KOH in ethanol, the free FAs were extracted with hexane and evaporated to dryness. Then, they were esterified (de Alaniz and Marra, 1992) , and the FA methyl esters (FAMEs) obtained were extracted twice with hexane. FAMEs were quantified by gas chromatography using a capillary-GLC-Hewlett Packard HP 6890 GC Series System Plus (Avondale, PA, USA) and were identified by comparisons of their relative retention times with authentic standards. The oven temperature was programmed to increase from 185 to 230 °C in increments of 3 °C/min. Ultra-dry helium was used as a carrier gas at an operating pressure of 3.0 kg/cm 2 . FAMEs were detected with a flame ionization detector operating at 280 °C. In some cases, the identities of the FAs were confirmed by mass spectrometry analysis. Each FA was expressed as μmol per mg of lipids in the sample.
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