High Performance Liquid Chromatographic Determination of Ascorbic Acid in Brassica Oleracea L. Var. Italica Plenck

M. Shekarchi¹, F. Khalili², Y. Mostofi²and H. Hajimehdipoor³*

¹Food and Drug Laboratory Research Center and Food and Drug Control Laboratories, MOH & ME, Tehran Iran.

²Department of Horticultural Sciences, College of Agriculture and Natural Resources, Tehran University, Karaj Iran.

³Traditional Medicne & Materia Medica Research Centre, Department of Traditional Pharmacy, Faculty of Traditional Medicine, Shahid Beheshti University of Medical Sciences, Tehran Iran.

Corresponding Author E-mail:hajimehd@tums.ac.ir

ABSTRACT: Numerous epidemiological studies indicate that broccoli protects human being against cancer. It is rich source of antioxidant materials specially flavonoids and vitamin C. In the present investigation, a reversed-phase high-performance liquid chromatographic procedure with UV detection at 245 nm has been developed and validated for the determination of vitamin C in broccoli. The freeze-dried tissues were extracted with mixture of acetic acid, meta-phosphoric acid, sulfuric acid and EDTA. Samples were centrifuged at 12000 rpm for 20 min at 4°C and 20 μl of this extract injected directly into the HPLC system. The HPLC system consisted of Lichrocart column C8 (250ª4.6 mm, 5 μm) and a gradient mode of potassium dihydrogen phosphate 50 mM : methanol which was kept at a flow rate of 0.5 ml/min. The method was validated for selectivity, linearity (r2>0.999), intra-day precision (£ 2.7), inter-day precision (£ 3.0) and accuracy measured by determination of recovery (96-106%). The LOD and LOQ were found 3.0 and 10.1 μg/ml, respectively. Because of speed and accuracy, it may be the UV-HPLC choice for analysis of vitamin C in Brassica oleracea L. var. italica Plenck

KEYWORDS: broccoli; HPLC; validation; vitamin C

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