Comparison between high-performance and ultra-high performance liquid chromatography methods for cholesterol determination in foods

Costa, H.S.; Oliveira, M.B.; Sanches-Silva, A.; Albuquerque, T.G.

http://hdl.handle.net/10400.18/1926

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Background and Objectives: Gas chromatography is still the most widely used technique for cholesterol determination, however liquid chromatography was shown to provide the most sensitive and accurate results. The purpose of this study was to compare high-performance and ultra-high performance liquid chromatography methods (HPLC and UHPLC) for cholesterol determination in foods. Methods: For HPLC method, a Supelcosil LC-18-DB (15 cm x 4.6 mm x 3.0 µm) column was used, whereas in UHPLC it was an Acquity BEH C18 (2.1 mm x 50 mm x 1.7 µm). For both methods, a UV detector (210 nm) and a mobile phase (acetonitrile/isopropanol, 70:30, v/v) were used. Methods were validated with respect to: linearity, limit of detection (LOD), limit of quantification (LOQ), recovery, accuracy and precision. Results: Both methods were suitable for cholesterol determination in foods, with linearity ranging from 0.005 to 0.40 mg/mL (HPLC - r2 = 0.9991; UHPLC – r2 =0.9992). However with UHPLC method, a lower LOQ (2.4 µg/mL) and LOD (0.73 µg/mL) and best recovery rates were achieved. Moreover, run time decreases from 4.5 min to 2 min. Also, it was possible to significantly reduce the amount of solvents used, since for UHPLC the flow rate is 0.3 mL/min instead of 1.2 mL/min. Conclusions: In this work, a new UHPLC method has been developed for quantification of cholesterol in foodstuffs. HPLC and UHPLC analysis have been compared. Analysis time, solvent consumption, and analysis cost are very important in many analytical laboratories. Overall, UHPLC can offer significant improvements compared with conventional HPLC.