Serum vitamins A and E deficiencies in patients with inflammatory bowel disease.

C inflammatory bowel diseases (IBD) include ulcerative colitis (UC) and Crohn’s disease (CD), the etiologies of which are not certainly defined, and are hypothesized to be an interaction between environmental, genetic, and immunologic factors.1 Recent studies show a gradually increasing rate of IBD in developing countries in Africa, South America, and Asia.1 There has been a reported increase in the incidence and prevalence of IBD in the West during the past 50 years, 120-200/100000 for ulcerative colitis and 50-200/100000 for Crohn’s disease (CD).1 Inflammatory bowel disease patients usually suffer from mal-absorption and mal-digestion, especially in the flare-up phase of the disease when increased requirement and decreased food intake are reported. Nutritional deficiency and malnutrition have been documented in IBD patients notably in CD, which may lead to adverse outcomes.2 Vitamins B12, A, E, and D need to be monitored and supplemented. When a large amount of the small bowel is resected, all nutrients will be absorbed poorly.3 This case-control study was designed to measure and compare serum levels of vitamins A and E between IBD patients and a healthy normal population in northeastern Iran. Due to the large differences in feeding habits in different areas, the aim of this study was to compare the levels of these vitamins in the patients and healthy cases. This case-control study was carried out from November 2011 to April 2012 in Golestan Province, Northern Iran. In this study, 94 pathologically confirmed IBD cases, and 94 healthy age and gendermatched controls with no gastrointestinal problems over the previous 2 years were recruited. The inclusion criteria for IBD patients were based on clinical diagnosis and pathology, and the exclusion criteria were based on those who did not receive vitamin supplements in the last 6 months. A fasting blood sample was taken. Measurement of vitamins A and E was carried out by the highperformance liquid chromatography (HPLC) technique (KNAUER V 7057-3; Smartline pump 1000 V 7603; Smartline UV Detector 2500 V 7604 and Smartline manager 5000 V 7602, (Berlin, Germany). Values of less than 30 μg/dl for vitamin A and 0.5μg/ dl for vitamin E were considered as deficiencies. Retinol acetate (Retinyl palmitate [all-trance-retinol palmitate]; Sigma LN 68F0645, St. Louis, MO, USA) was used as the internal standard with a final concentration of 2 μg/ml. To extract vitamins, 200 μl methanol (Merck & Co., Inc., Rahway, NJ, USA), 200 μl Ethanol (Merck & Co., Inc., Rahway, NJ, USA, 11727), 50 μl internal standard (Retinol acetate) and 500 μl n-Hexan (Merck & Co., Inc., Rahway, NJ, USA, 04391) were added to 200 μl of plasma. The surface phase was gathered after centrifuge and 500 μl n-Hexan added, and then vaporized by nitrogen gas. The remaining was dissolved in 200 μl methanol, and 50 μl of sample was injected for HPLC (flow rate: 1.5 ml/min, time: 20 min, sample: 50μl, wavelength [0 min–7 min]: 325 nm, wavelength [7 min -15 min]: 292 nm)]. The ethical committee of Golestan University of Medical Sciences (No=1453) approved this study, and the patient consents were obtained. Fifty percent of both case and control groups were male. The mean (standard deviation [SD]) age of patients was 38 (15), and of controls was 38 (14) years (p=0.8).