Biocatalysis and Agricultural Biotechnology 17 (2019) 416–421 Contents lists available at ScienceDirect Biocatalysis and Agricultural Biotechnology journal homepage: www.elsevier.com/locate/bab Phytochemical composition and antioxidant characteristics of traditional T cold pressed seed oils in South India ⁎ Devanesan Arul Anantha, , Garlapati Deviramb, Vijayaraghavan Mahalakshmic,d, ⁎ Thilagar Sivasudhad, Zipora Tietela, a Department of Food Quality and Safety, Gilat Research Center, Agricultural Research Organization, M.P. Negev 85280, Israel b National Centre for Coastal Research (NCCR), Ministry of Earth Sciences (MoES), Pallikaranai, Chennai 600100, Tamil Nadu, India c AU-KBC Research Centre, MIT campus, Anna University, Chennai, India d Department of Environmental Biotechnology, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India ARTICLE INFO ABSTRACT Keywords: Cold press extraction is the ultimate method to extract essential phytonutrients from oil seeds. Other extractions Seed oil such as chemical and hot press extraction include processes like deodorizing and refining which affect the Tocopherols quality of phytocompounds (essential fatty acids, tocopherols and phytosterols). In this study, antioxidant ca- Fatty acids pacity, tocopherol and fatty acid profiles of six cold-pressed oils (sesame, coconut, peanut, neem, castor and UPLC iluppai) were determined. Lipophilic extracts showed excellent DPPH-free radical scavenging activity compared GC-MS to hydrophilic extracts. Lipophilic extract of sesame oil showed the highest antioxidant capacity in both DPPH (0.61 ± 0.02 TEAC µM/g dw) and ABTS (0.85 ± 0.05 TEAC µM/g dw) free radical scavenging assays. The presence of alpha (α), gamma (γ) and delta (δ) tocopherols were detected and quantified in all the six seed oil samples through UPLC analysis. From the results it was confirmed that γ- tocopherol content is predominant in castor (395.32 ± 2.12 mg/kg), sesame (278.72 ± 2.80 mg/kg) and peanut (104.11 ± 2.86 mg/kg) oils. GC- MS analysis of all the six seed oil samples exhibited fatty acids such as myristic, palmitic, stearic, oleic and linoleic acid in various compositions. The highest levels of total saturated (SFA) in coconut oil, monounsaturated (MUFA) in castor oil and polyunsaturated (PUFA) in sesame oil were quantified at the range of 85.51%, 64.09%, and 31.72% respectively. In conclusion these phytoconstituents from cold pressed oil play a promising role in improving human health without side effects. 1. Introduction classified into ω−3, ω−6 and ω−9 fatty acids. Omega-3 &6(ω−3 and ω−6) are called essential fatty acids, because they are acquired Plant edible oil is a routine source for food processing to improve only through foods consumption and cannot be synthesized by mam- the food taste, flavor and texture in addition its other medicinal ap- mals on their own. A pilot study on major depressive disorder (MDD) plications. Commonly utilized oil seed crops for extraction are sesame, and cardiovascular diseases patients showed strong association with mahua, olive, soybean, corn, peanut, cotton, palm nuts, sunflower, polyunsaturated fatty acids, and the authors concluded that it confirms neem, to name a few. In general, these edible oils are acquired from the its preventive role in these diseases (Assiesa et al., 2004). On the other inner part (endosperm) of seed and fruit (pericarp). A report showed an hand, unsaturated fatty acids like linoleic and linolenic acids sig- upsurge in global consumption of vegetable oil, with 177 million metric nificantly increases HDL-cholesterol (good cholesterol) and decrease tons having been produced in 2015–2016. Moreover, sunflower seed oil LDL-cholesterol (bad cholesterol). Similarly, oleic acid decreases LDL- consumption in particular amounted to 14.91 million metric tons cholesterol without affecting HDL cholesterol levels (Lichtenstein et al., (USDA, 2017). Edible seed oil contains saturated (SFA), mono- 2006). According to the American Heart Association report, the "bad" unsaturated (MUFA) and polyunsaturated (PUFA) fatty acids, and other cholesterol levels (low-density lipoprotein) significantly decreased due valuable bioactive substances such as tocopherols and tocotrienols, to the intake of nutrient-rich edible oil. Other unsaturated fatty acids, polyphenols, phytosterols, lignans, triterpene, carotenoids and chlor- which are naturally present in seed oil have excellent antioxidant ca- ophylls (Ghanbari et al., 2012). Polyunsaturated fatty acids are further pacity and protect the organism from free radical damage and act ⁎ Corresponding authors. E-mail address: [email protected] (Z. Tietel). https://doi.org/10.1016/j.bcab.2018.12.018 Received 23 October 2018; Received in revised form 15 December 2018; Accepted 16 December 2018 Available online 18 December 2018 1878-8181/ © 2018 Elsevier Ltd. All rights reserved. D.A. Ananth et al. Biocatalysis and Agricultural Biotechnology 17 (2019) 416–421 against cardiovascular and degenerative diseases (Gamazo-Vázquez 2.2. Oil samples et al., 2003). Thereupon edible oil unsaturated fatty acids play a pivotal role in lipid metabolism and essential in human diet. Sesame, coconut, neem, peanut, castor and iluppai seeds were col- Oil seed extraction can be carried out either by hot or cold oil lected from rural markets in Usilambatti, Tamil Nadu, India. Seeds were pressing methods. The method of extraction depends on the source and dried in the sun and extracted using traditional cold pressing stones the type of material to be recovered in the final oil yield. During hot oil called “Chekku”. Samples were kept in airtight screw cap containers at extraction, essential nutrients and bioactive compounds are degraded room temperature for further analysis. (Garcia et al., 2006). Essential fatty acids and nutrients like vitamin E are present in high level in cold pressed edible oils (Butinar et al., 2.3. Hydrophilic and lipophilic extraction 2011), which also preserve bioactive phytochemicals, essential nu- trients and antioxidants. In recent years, cold pressing has become the Oil samples (0.3 g) were diluted with 300 µl of 80% methanol (1:1, more desirable method for oil extraction from oil seeds and fruits. This v-v) and 326 µl of n-hexane. After centrifuging at 17,000xg for 6 min, process is also very user friendly and inexpensive when compared to hydrophilic and lipophilic fractions were separated and used for further other commercial extractions. On the other hand, even though the su- analysis (Miniotia, Georgioua, 2010). These extracts were used only for percritical carbon dioxide (CO2) extraction method is one of the most antioxidant analysis of seed oil. ecofriendly methods known for oil extraction, the cost of the equipment and other oil pressing materials is very expensive as compared to cold 2.4. ABTS free radical scavenging activity press oil extraction (Nam and Nam, 2016). Cold oil-pressing of seeds and fruits such as sunflower, olive, sesame, canola, coconut, and ra- ABTS free radical scavenging activity was carried out by the method peseed oil is now common, and these can all be extracted without described in Araki et al. (1999) with slight modification. ABTS radical solvents and hence retain the oil's quality as well as its natural flavor cations were generated in acidified ethanol medium to allow the per- and taste. Nowadays consumers prefer healthy beneficial foods, rich in formance of both hydro-lipophilic antioxidants. The reaction mixture natural and nutritionally safe phytonutrients. to maintain the immune contained 0.15 mM ABTS and 1.77 mM of K2S2O8 in acidified ethanol. system by preventing disease. Due to ageing, environment, changes in The reaction mixture was incubated at 45 °C for 60 min for ABTS+ food consumption (alcohol, artificial additives), emotional and physical radical generation in a water bath. One ml of ABTS+ solution was stress, free radicals are generated and irreversibly accumulated. Free added to 10 µl of oil sample and incubated for 30 min at 25 °C. Radical radicals induce cell and tissue damage by inactivating biomolecules like scavenging activity was spectrophotometrically measured at 734 nm. proteins, lipids and DNA, which in turn lead to many degenerative Trolox was used as a standard and the radical scavenging activity of diseases. Antioxidants have the capacity to neutralize free radicals and both hydrophilic and lipophilic fractions were expressed as Trolox prevent the cell from oxidative damage. Cold pressed seed oil extracts equivalent antioxidant capacity per gram of oil dry weight of the have the potential to protect the human immune system as they have sample (TEAC/g dw). abundant natural antioxidant compounds (Yu et al., 2005; Teh, Morlock, 2015; Topkafa, 2016). These oils are a rich source of vitamin 2.5. DPPH free radical scavenging activity E, unsaturated fatty acids (oleic, linoleic and α-linoleic acid) and li- pophilic antioxidants such as α, γ and δ tocopherols which primarily The method described in Tuberoso et al. (2007) was adopted for the contribute the total antioxidant capacity of the cold pressed oils (Carlo determination of DPPH free radicals scavenging activity of oil samples. et al., 2007). In comparison to hot pressed, cold pressed extract showed One ml of DPPH (0.04 mM) solution was prepared in methanol and superior antioxidative properties (lipophilic antioxidants) (Lerma- mixed with hydrophilic and lipophilic oil fractions (10 µl). After one Garcia et al., 2009). Subsequently, cold pressed