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Simultaneous HPLC quantification of total cholesterol,tocopherols and β-carotene in Barrosã-PDO veal
Authors:José A Mestre Prates  Mário A Gonçalves Quaresma  Rui J Branquinho Bessa  Carlos MG Andrade Fontes  Cristina MP Mateus Alfaia
Affiliation:1. Department of Biochemistry, Faculty of Veterinary Medicine – CIISA, Rua Professor Cid dos Santos, Pólo Universitário do Alto da Ajuda, 1300-477 Lisboa, Portugal;2. Estação Zootécnica Nacional, Instituto Nacional de Investigação Agrária e das Pescas, Fonte Boa, 2005-048 Vale de Santarém, Portugal
Abstract:A simple, rapid and sensitive procedure for the simultaneous determination of total cholesterol, tocopherols and β-carotene in meat is described. The method involves a direct saponification of the meat, a single n-hexane extraction and the analysis of the extracted compounds by normal-phase HPLC, using fluorescence (tocopherols) and UV–Vis photodiode array (cholesterol and β-carotene) detections in tandem. Rates of recovery of spiked meat samples were 93% for cholesterol, 83–86% for (α-, β- and -γ) tocopherols and 89% for β-carotene. Repeatabilities were high (CV < 6%) for all determined compounds, except for δ-tocopherol. This tocopherol, which is not usually present in meat, showed a much lower recovery percentage (73%) and repeatability (12.8%). This methodology was applied for the quantification of total cholesterol, tocopherols and β-carotene in three muscles (longissimus thoracis, longissimus lumborum and semitendinosus) of the Portuguese traditional Barrosã-PDO veal, obtained from autochthonous calves fed extensively during summer (with the least abundant green pastures) and slaughtered in early autumn (October). Barrosã-PDO veal showed median contents of total cholesterol (0.50–0.56 mg/g) and, depending on the analysed muscle, moderate to high contents of α-tocopherol (3.3–3.9 μg/g) and β-carotene (0.07–0.09 μg/g), suggesting an high sensorial and hygienic quality.
Keywords:ANOVA  analysis of variance  BHT  2  4-di-tert-butyl hydroxytoluene  CHR  cholesterol  β-CT  β-carotene  CV  coefficient of variation  HPLC  high-performance liquid chromatography  LL  longissimus lumborum  LT  longissimus thoracis  LSD  least significant difference  PDO  protected designation of origin  ST  semitendinosus  TF  tocopherols  UV  ultraviolet
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