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Download Download Available online: www.notulaebotanicae.ro Print ISSN 0255-965X; Electronic 1842-4309 Not Bot Horti Agrobo, 2017, 45(1):179-184. DOI:10.15835/nbha45110678 Original Article Bioactive Compounds, Antioxidant Activity and Nutritional Quality of Different Culinary Aromatic Herbs Violeta NOUR1*, Ion TRANDAFIR2, Sina COSMULESCU1 1University of Craiova, Department of Horticulture and Food Science, 13 A.I. Cuza Street, 200585 Craiova, Romania; [email protected] (*corresponding author); [email protected] 2University of Craiova, Department of Chemistry, 107 Calea Bucuresti Street, 200529 Craiova, Romania; [email protected] Abstract In the present work, four of the most widely used culinary herbs (parsley, dill, lovage and celery leaves) in the Romanian traditional cuisine were assessed for some nutritional quality parameters (moisture content, titratable acidity, ascorbic acid content and mineral content), total phenolics content, total flavonoids content and antioxidant activity by 1,1-diphenyl-2- picrylhydrazyl (DPPH) radical scavenging assay. In addition, high-performance liquid chromatography (HPLC) coupled with diode-array detection was applied for the identification and quantification of some individual phenolic compounds in the selected aromatic herbs. Although parsley showed the highest ascorbic acid content (206.32 mg/100 g fw), it registered the lowest antioxidant activity. Fresh lovage recorded the highest total phenolic content (577.04 mg GAE/100 g), total flavonoids content (298.38 mg QE/100 g) and antioxidant activity, followed by dill and celery leaves. Among the identified phenolic acids, ellagic acid was predominant in lovage and celery leaves whereas sinapic and vanillic acids were prevalent in dill. Among flavonoids, myricetin was predominant in parsley and celery leaves whereas rutin was predominant in dill and lovage. Keywords: antioxidant activity, ascorbic acid, celery leaves, dill, flavonoids, lovage, parsley, phenolic acids The bioactive compounds, which are the key natural flavour and fragrance ingredients of various spices and other aromatic plants, play a vital role in healthcare (Gunasekar et al., Introduction 2012). Herbs and spices are a dietary source of bioactive polyphenols (Hinneburg et al., 2006), which are now known Spices are essentially used to improve the appeal and to possess beneficial effects for human health, including sensory characteristics of food. Extensive experiments have digestive stimulant, anti-inflammatory, antimicrobial, proved the health benefits of spices without any deleterious antioxidant and anticarcinogenic activities (Shobana and side-effects (Gunasekar et al., 2012). The aromatic herbs used Akhilender Naidu, 2000; Zheng and Wang, 2001). In addition in flavouring food and beverages are also sometimes included as to phenolic compounds, these aromatic herbs contain spices. They have been grown and used for hundreds of years, numerous phytochemicals such as nitrogen compounds, carotenoids, and ascorbic acid. Many of these phytochemicals and they are becoming increasingly popular for their ability to enhance and complement the flavors of a wide variety of foods possess significant antioxidant capacities that are associated (Zheng and Wang, 2001). Culinary aromatic herbs are good with lower incidence and lower mortality rates of cancer in sources of essential nutrients such as vitamins and minerals as several human cohorts (Zheng and Wang, 2001). Moreover, well as antioxidant compounds. Supplementation of diet with the volatile constituents (essential oils) that are the main cause for use of these plants can significantly contribute to biological various herbs is recommended among individual consumers, both for its healing properties and nutritive value (Yen et al., activity (Inouye et al., 2001). 2002). Numerous studies have shown that a high intake of Recently, there has been growing awareness of the vegetables that are sources of antioxidant compounds is importance of a high dietary content of phenolic compounds, associated with a lower incidence of cardiovascular disease, such as flavonoids and hydroxycinnamic acids (Vallverdu- cancer and other chronic diseases. The relatively low intake Queralt et al., 2014), because of their apparent multiple levels of culinary herbs and spices do not necessarily mean that biological effects, including metal chelation, free-radical they are of little value as their high polyphenol content, and scavenging, inhibition of cellular proliferation, modulation of thus ultimately the potential biological impact of this content, enzymatic activity and signal transduction pathways (Del Rio et cannot be ignored (Opara and Chohan, 2014). al., 2013). Received: 04 Jan 2017. Received in revised form: 06 Apr 2017. Accepted: 07 Apr 2017. Published online: 13 Apr 2017. Nour V et al / Not Bot Horti Agrobo, 2017, 45(1):179-184 180 In recent years, the search of natural and safe antioxidants, system equipped with a Diode Array Detector (DAD) and a reverse-phase C18 column (Hypersil Gold aQ, 25 cm × 4.6 mm, especially of plant origin has increased considerably, mainly because synthetic antioxidants such as butylated hydroxyl- 5 μm) maintained at 10 °C. Elution (flow rate of 0.7 mL/min) toluene (BHT) or butylated hydroxyanysol (BHA) have been was performed in isocratic conditions. The mobile phase confirmed or suspected to be associated with potential health consisted of a phosphate buffer solution (50 mM KH2PO4 in hazards (Hinneburg et al., 2006). Therefore, searching for water) adjusted to a pH value of 2.8. The absorbance was natural antioxidants from herbs or plants, to attenuate measured at 254 nm and the ascorbic acid peak area was oxidative and nitrosative stress, is of growing interest (Wu et al., quantified with the Chrom Quest 4.2 software on the basis of an 2011). external standard calibration curve (0-50 mg/L). Ascorbic acid In this study there were selected four widely used aromatic content was expressed as mg/100 g fresh weight (fw). herbs in the traditional Romanian cuisine, which are known to possess strong pharmacological activity and high antioxidant Minerals content content: dill, parsley, lovage and celery leaves. They were tested Calcium (Ca), magnesium (Mg), sodium (Na) and for their content of biologically active compounds (ascorbic potassium (K) were determined by microwave digestion acid, phenolic compounds, and flavonoids) and their followed by inductively coupled plasma mass spectrometry (ICP- antioxidant activity. Another purpose of this study was to MS). Samples of approximately 2.0 g were weighed in TFM indentify and quantify some individual phenolic compounds in vessels, over that 5 mL nitric acid 65% and 2 mL hydrogen these aromatic herbs using high performance liquid peroxide 30% were added. The vessels were then closed and chromatography (HPLC) coupled with diode-array detection. heated at 180 °C for 20 min in the Milestone digestion microwave system. Reagent blanks were included in each series of digestions. After digestion was completed, the clear, colourless Materials and Methods solution was transferred into a 50 mL volumetric flask and diluted to the mark with ultrapure water. The sample solutions Chemicals and reagents were stored in polyethylene vials at 6 °C until analysis. An Elan Folin-Ciocalteu reagent (2 N), 6-hydroxy-2,5,7,8- 9000 inductively coupled plasma mass spectrometer (Perkin tetramethylchroman-2-carboxylic acid (Trolox), HPLC grade Elmer Sciex, Canada) equipped with Meinhard nebulizer and acetonitrile, acetone, n-hexane and methanol were purchased Scott-type double-pass spray chamber was used throughout. from Merk (Germany). Gallic acid (99% purity), anhydrous Calibration standards were prepared by serial dilution of sodium carbonate (99% purity), aluminium nitrate, potassium commercially prepared analytical standard solutions acetate, acetic acid, quercetin and 2,2-diphenyl-1-picrylhydrazyl (PerkinElmer, Shelton, CT) and linear calibration curves were (DPPH, 90% purity) were from Sigma-Aldrich (Germany). established for concentration vs. absorbance. The results were Standards of phenolic acids (gallic, vanillic, chlorogenic, caffeic, expressed in mg per 100 g of fresh weight. syringic, p-coumaric, ferulic, sinapic, salycilic, ellagic and trans- cinnamic) and flavonoids (catechin, epicatechin, rutin, myricetin Extraction of phenolic compounds and quercetin) were also purchased from Sigma-Aldrich. For the analysis of total phenolics, total flavonoids and Ultrapure water was obtained from a Milli-Q water purification antioxidant activity, samples (1.5 g) were extracted with 10 mL system (TGI Pure Water Systems, USA). methanol in a Bandelin Sonorex Digital 10P ultrasonic bath (Bandelin Electronic GmbH, Germany) for 60 min at room Plant material temperature. Supernatants were collected, filtered through 0.45 Fresh plant materials of dill (Anethum graveolens), parsley μm polyamide membranes and used for the assays. (Petroselinum crispum), lovage (Levisticum officinale) and celery (Apium graveolens L.) leaves were purchased from a local market Total phenolics content in Craiova, Romania. Samples were washed under tap water and The total phenolics content was assessed by using the Folin- inedible parts were removed. Samples were frozen at −20 °C Ciocalteu phenol reagent method according to the method of until analysis. For quality parameters analyses (moisture content, Singleton and Rossi (1965) with minor modifications titratable acidity, ascorbic acid content), sub-samples were stored (Cosmulescu
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