eng Oriental Scientific Publishing Company Biosciences Biotechnology Research Asia 0973-1245 2016-05-05 8 1 95 100 9247 article Shekarchi 1 F. Khalili 2 Y. Mostof 2 H. Hajimehdipoor 3 Food and Drug Laboratory Research Center and Food and Drug Control Laboratories, MOH & ME, Tehran (Iran). Department of Horticultural Sciences, College of Agriculture and Natural Resources, Tehran University, Karaj (Iran). Traditional Medicne & Materia Medica Research Centre, Department of Traditional Pharmacy, Faculty of Traditional Medicine, Shahid Beheshti University of Medical Sciences, Tehran (Iran). 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 https://www.biotech-asia.org/vol8no1/high-performance-liquid-chromatographic-determination-of-ascorbic-acid-in-brassica-oleracea-l-var-italica-plenck/ broccoli; HPLC; validation; vitamin C