Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials
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...
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2020-09-01
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doaj-00a30fc7f858491d8578bc155e4f79442020-11-25T02:11:37ZengMDPI AGFoods2304-81582020-09-0191378137810.3390/foods9101378Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference MaterialsLukáš Kolarič0Peter Šimko1Faculty of Chemical and Food Technology, Institute of Food Science and Nutrition, Slovak University of Technology in Bratislava, Radlinského 9, 812 37 Bratislava, SlovakiaFaculty of Chemical and Food Technology, Institute of Food Science and Nutrition, Slovak University of Technology in Bratislava, Radlinského 9, 812 37 Bratislava, SlovakiaThis 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, <i>v/v</i>) double extraction. HPLC separation consisted of isocratic elution with flow rate of 0.5 mL/min mobile phase composed of acetonitrile/methanol 60:40 (<i>v/v</i>) 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.https://www.mdpi.com/2304-8158/9/10/1378cholesterolbuttersaponificationextractionHPLCoptimization |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Lukáš Kolarič Peter Šimko |
spellingShingle |
Lukáš Kolarič Peter Šimko Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials Foods cholesterol butter saponification extraction HPLC optimization |
author_facet |
Lukáš Kolarič Peter Šimko |
author_sort |
Lukáš Kolarič |
title |
Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials |
title_short |
Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials |
title_full |
Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials |
title_fullStr |
Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials |
title_full_unstemmed |
Determination of Cholesterol Content in Butter by HPLC: Up-to-Date Optimization, and In-House Validation Using Reference Materials |
title_sort |
determination of cholesterol content in butter by hplc: up-to-date optimization, and in-house validation using reference materials |
publisher |
MDPI AG |
series |
Foods |
issn |
2304-8158 |
publishDate |
2020-09-01 |
description |
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, <i>v/v</i>) double extraction. HPLC separation consisted of isocratic elution with flow rate of 0.5 mL/min mobile phase composed of acetonitrile/methanol 60:40 (<i>v/v</i>) 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. |
topic |
cholesterol butter saponification extraction HPLC optimization |
url |
https://www.mdpi.com/2304-8158/9/10/1378 |
work_keys_str_mv |
AT lukaskolaric determinationofcholesterolcontentinbutterbyhplcuptodateoptimizationandinhousevalidationusingreferencematerials AT petersimko determinationofcholesterolcontentinbutterbyhplcuptodateoptimizationandinhousevalidationusingreferencematerials |
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