Biological antioxidant assays can be a more useful approach compared to chemical assays as they can provide details on the ability of antioxidants to prevent oxidation of biological materials such as lipids, DNA and proteins. In this study, isolated serum, LDL and haemoglobin were used as the biological models to measure the ability of the water extracts of the leaves and stems of B. racemosa to prevent their oxidation. We hope

this current study can shed more light on the ability of B. racemosa extracts to act as anti-oxidative agents based on biological materials. This study is a continuation of our previous work, focusing on the aqueous extracts of B. racemosa which, when compared to the ethanol, ethyl acetate and hexane extracts, possess the highest phenolic content and antioxidant VE-821 ic50 activities, ( Kong et al., 2012). We quantified the polyphenols, both in the free and bound forms, to better understand the nature of their structure in the plant, which would shed some light on CX-5461 order their bioavailability and subsequent bioactivity. In addition, we tested the effects of two drying methods (freeze drying vs. air drying) on the polyphenolic content of the plant. An optimal drying method

is important to prevent losses of polyphenols during processing. HPLC-grade and other analytical grade chemicals and reagents were obtained from general suppliers. Diethyldithiocarbamic acid (DETC) sodium salt and all polyphenolic standards were of HPLC grade (purity > 95%) and were obtained from Sigma–Aldrich Chemical Co. (St. Louis, MO). The shoots of B. racemosa were collected from the state of Kedah, in northern Peninsular Malaysia. The leaves and stems of the shoots were separated Anacetrapib and dried using two methods: freeze drying and air drying. For freeze drying, samples were allowed to freeze at −80 °C for 3 days before lyophilisation in a freeze

dryer. Air drying was performed under a running fan at room temperature for 7 days. The dried stems and leaves were subsequently ground into powder and sieved via a 1-mm mesh. All prepared samples were stored at −20 °C until further analyses. Two grams of either freeze-dried or air-dried samples were extracted with 40 ml of water. Each extraction was performed three times, in an incubator shaker (Innova 4300; New Brunswick Scientific, New Jersey, USA) at 200 rpm and 30 °C for 24 h. The extract was later centrifuged (Jouan CR3i multifunction centrifuge; Thermo Scientific, Waltham, NJ) at 1389g for 5 min at 4 °C. The supernatant was filtered through Whatman No. 4. paper and the filtrate was lyophilised. The dried powder was weighed and pooled together and stored at −20 °C until further analyses. Free polyphenols (X) were estimated by dissolving the dried powder in water containing 20 mM DETC sodium salt prior to UHPLC analysis.