Sugars, Ascorbic Acid, Total Phenolic Content and Total Antioxidant Activity
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Research Article Received: 16 May 2012 Revised: 3 August 2012 Accepted article published: 20 August 2012 Published online in Wiley Online Library: 2 October 2012 (wileyonlinelibrary.com) DOI 10.1002/jsfa.5876 Sugars, ascorbic acid, total phenolic content and total antioxidant activity in passion fruit (Passiflora) cultivars Shiamala Devi Ramaiya,a∗ Japar Sidik Bujang,a Muta Harah Zakaria,b Wong Sing Kinga and Muhd Arif Shaffiq Sahrira Abstract BACKGROUND: The levels of sugars, ascorbic acid, total phenolic content (TPC) and total antioxidant activity (TAA) were determined in fruit juices from seven passion fruit (Passiflora spp.) cultivars: P. edulis cultivars Purple, Frederick, Yellow, Pink, P. edulis f. flavicarpa, P. maliformis and P. quadrangularis (we also tested this cultivar’s mesocarp). RESULTS: Purple and Yellow P. edulis had significantly higher total sugar, 142.85 ± 0.17 g kg−1 and 139.69 ± 0.12 g kg−1, respectively, than other cultivars. Glucose and fructose content were higher in juice from vine-ripened fruits of Purple, Frederick and Yellow P. edulis, P. quadrangularis and P. maliformis. Sucrose content was significantly higher in juice of non-vine-ripened fruits of P. edulis (Pink) and P. edulis f. flavicarpa. Ascorbic acid, TPC and TAA were significantly higher in vine-ripened Purple and Yellow P. edulis; ranges were 0.22–0.33 g kg−1, 342.80–382.00 mg gallic acid equivalent L−1 and 409.13–586.70 µmol Trolox ◦ L−1, respectively. Based on principal component analysis (PCA) and cluster analysis, the main variables – Brix, total sugar, glucose, fructose, ascorbic acid, TPC and TAA – formed the characteristics for the group comprising Purple and Yellow P. edulis. CONCLUSIONS: Glucose, fructose, sucrose, ascorbic acid, TAA and TPC were quantified in passion fruit juices. Variation of the above variables in juices of Passiflora depends on the cultivar and ripeness. c 2012 Society of Chemical Industry Keywords: ascorbic acid; HPLC; passion fruit cultivars; sugars; total antioxidant activity; total phenolic content INTRODUCTION nutrients content including micronutrients such as minerals, fiber, 11 The passion fruit is an exotic fruit that is popular for the startling phenolic compounds and ascorbic acid content. A number of beauty of its flower and fruity aroma.1 It enters the international studies on other plant species (e.g. cashew apple, sweet cherry, trade mainly in the form of juice. The world production of passion strawberries, oranges) indicate the importance of micronutrients 12–16 fruit increased from approximately 1.05 million Mt in 2005 to for human health. 1.27 million Mt in 2010, with Brazil as the larger producer and The sugar content in fruit can influence the physiochemical consumer of passion fruit in the world.2 Other important passion properties(e.g.pH,totalacidity,microbialstability)andcanprovide 17 fruit producing countries are Ecuador, Colombia, Peru, Australia, valuable information on food wholesomeness. The predominant New Zealand, South Africa and the state of Hawaii.3 In Peninsular sugars in passion fruit juices are sucrose, glucose and fructose, Malaysia, passion fruit has been grown in Johor and the Cameron which are of economic importance. In passion fruit the ascorbic Highlands since pre-war days and fruit production in these regions acid content is lower compared to the sugar content and it 9 has been affected by a viral disease which discouraged further affects the characteristics of fruit quality. Phenolic compounds expansion in commercial planting.4 However, this fruit is still are secondary metabolites naturally constituent in many fruits and cultivated in Peninsular Malaysia owing to the prevalence of well known for their antioxidant potential, food quality and their suitable growing conditions and increase in demand. role in prevention of human diseases, and reducing the risk of The physical and chemical composition of passion fruit can vary according to cultivar, environmental factors including climate and soil condition, and agricultural practices.5 Harvesting time ∗ Correspondence to: Shiamala Devi Ramaiya, Department of Animal and postharvest are also factors that may affect the fruit’s Science and Fisheries, Faculty of Agriculture and Food Sciences, Universiti 5,6 Putra Malaysia Bintulu Sarawak Campus, 97008 Bintulu, Sarawak. E-mail: sensory and nutritional quality. Quantitative information [email protected] concerning the fruit’s content of sugars, ascorbic acid, organic acids and phenolics is the most important factor in verifying a Department of Animal Science and Fisheries, Faculty of Agriculture and Food the authenticity and quality index for fruits used in food Sciences, Universiti Putra Malaysia, 97008, Bintulu, Sarawak 1198 7–10 manufacturing. Demands for passion fruit juice are increasing b Department of Aquaculture, Faculty of Agriculture, Universiti Putra Malaysia, not only because of the juice’s exotic flavor but also its essential 43400, UPM Serdang, Selangor Darul Ehsan, Malaysia J Sci Food Agric 2013; 93: 1198–1205 www.soci.org c 2012 Society of Chemical Industry Chemical composition and antioxidant activity of Passiflora cultivars www.soci.org cancer and cardiovascular disease.18 The antioxidant activity in analysis. Prior to injection all juices were filtered through a 0.45 passion fruit is notable since the fruit is rich in compounds that µm pore size membrane filter (Waters-Millipore Corp., Milford, MA, function in free radical scavenging activity.19 USA) and 10 µL were used per injection. Thesugarsinpassionfruithavebeeninvestigatedbyresearchers from various geographical locations using different approaches on Calibration standards the most important commercial passion fruit cultivars: Passiflora For quantitative examination of sugar content in the juices, 1,11,20,21 22,23 edulisf.flavicarpa andP.edulisSims. Eventhoughawide standard solutions of HPLC-grade ≥99.5% (Sigma, Steinheim, range of studies have been published on the sugar and organic Germany) sucrose, glucose and fructose were prepared in mobile acid contents, few have made use of the high-performance liquid phase. Calibration standards ranging from 0.05 to 1.0 g L−1 were 17,24,25 chromatography (HPLC) technique for fruit juice analysis. made for each sugar (sucrose: R2 = 0.9998, limit of detection (LOD) More attention has also been drawn to ascorbic acid, phenolic = 0.0218 mg kg−1, limit of quantitation (LOQ) = 0.0727 mg kg−1; 1,19,21,26 and antioxidant activity in passion fruit. Several studies glucose: R2 = 0.9999, LOD = 0.0095 mg kg−1,LOQ= 0.0316 mg have highlighted a strong correlation between total antioxidant kg−1;fructose:R2 = 0.9999, LOD = 0.0085 mg kg−1,LOQ= 0.0283 21,26,27 activity (TAA) and total phenolic content (TPC). However, mg kg−1) to determining the concentrations. very little information on sugars, ascorbic acid, phenolic content and antioxidant activity is available in the literature for various cultivars of passion fruit. HPLC components and operating conditions The aim of this study was to determine the sugars, ascorbic The complete HPLC system (Waters Corp., Milford, MA, USA) acid, total phenolic content and total antioxidant activity of fruit was equipped with a Waters Delta 600 pump controller, an juices from passion fruit cultivars to account for taste differences. in-line Waters degasser AF connected with a Waters 2414 Therefore, evaluating their chemical constituents and antioxidant refractive index detector. A Waters 2707 autosampler was used to activity can help in understanding the worth of these fruits in introduce a sample automatically into the inlets. An ion exclusion processing them as juice and other processed products. chromatography Sugar-Pak I column (Waters Corp.) was used to separated the soluble sugars. The 6.5 × 300 mm Sugar Pak I column was packed with a microparticulate cation-exchange gel EXPERIMENTAL in calcium form. The juices were isocratically separated through the column at a flow rate of 0.4 mL min−1. The temperature of Fruit harvest and storage ◦ Two to four kilograms (∼30–60 fruits) of vine-ripened fruits the detector and column was set at to 40 and 70 C, respectively. (based on the fruit’s skin color and firmness) of P. edulis (Purple), Sugar compounds in juices were identified by comparing their P. edulis (Frederick), P. maliformis and about 25 kg (16 fruits) retention times with standard sugars and quantified by peak area P. quadrangularis were harvested randomly from 10 plants for measurement using Waters Empower Pro software. each cultivar at the passion fruit farm, Universiti Putra Malaysia Bintulu Sarawak Campus (UPMKB) Bintulu, Sarawak, during the Determination of ascorbic acid content fruit-bearing season in April–June of 2011. Vine-ripened fruits of Juices were extracted from fresh passion fruit pulp, filtered through P. edulis (Yellow) were collected from an orchard at Ba’kelalan, Whatman no. 2 filter paper (Camlab, Cambridge, UK) and used for Sarawak, while non-vine-ripened fruits of P. edulis (Pink) and P. ascorbic acid analysis. Ascorbic acid was determined according edulis f. flavicarpa were picked from a farm at Sarikei, Sarawak. The to Suntornsuk et al.29 and Silva et al.30 with some modification fruits were brought to the laboratory and immediately inspected using a direct titration method. Ascorbic acid (Merck, Whitehouse and cleaned. Fruits were cut in half and the pulps were scooped Station, NJ, USA) was used as standard. The filtered juice (25 mL) out with a spoon. The pulps were separated from the seeds using a was titrated against iodine solution until the endpoint blue-black ◦ 2.0 mm pore size sieve. The pulps, either fresh or stored at −20 C color developed. The volume of iodine solution required to reach were subsequently used for the various analyses described below. the endpoint was recorded. Titrations were performed in triplicate for standards and filtered juice of passion fruit. General juice content ◦ Total soluble solid ( Brix), pH and total titratable acidity (TTA) Extraction for TPC and TAA The extracted juices from fruit pulps were analyzed for soluble Passion fruit pulp (4 mL) was extracted with 40 mL solution solids content by the index of refraction, pH and TTA.