Content and composition of lipid classes, fatty acid from Sargassum seaweed collected at Con Dao and Van Phong bay

Nội dung text: Content and composition of lipid classes, fatty acid from Sargassum seaweed collected at Con Dao and Van Phong bay

1. Vietnam Journal of Marine Science and Technology; Vol. 21, No. 2; 2021: 311–318 DOI: Content and composition of lipid classes, fatty acid from Sargassum seaweed collected at Con Dao and Van Phong bay Thu Hue Pham1,2, Van Tuyen Anh Nguyen3, Thi Hoang Yen Kieu3, Le My Anh Nguyen, Hai Nam Hoang3, Quoc Long Pham3, Duc Tien Dam4,5, Thi Thanh Van Tran6, Tat Thanh Le1,3,* 1Graduate University of Science and Technology, VAST, Vietnam 2Vietnam Naval Academy, Nha Trang, Vietnam 3Institute of Nature Products Chemistry, VAST, Vietnam 4Institute of Marine Environment and Resources, VAST, Vietnam 5Haiphong University of Medicine and Pharmacy 6Nha Trang Institute of Technology Research and Application, VAST, Vietnam *E-mail: thanh.biotech@gmail.com Received: 26 November 2020; Accepted: 1 February 2021 â2021 Vietnam Academy of Science and Technology (VAST) Abtract This study studied the content and composition of the total lipid, lipid classes and fatty acids in 13 brown seaweed Sargassum species collected from Con Dao and Van Phong, Vietnam. The total lipid has a low content and varies among species from 0.10–1.70% of the fresh weight. From 13 species, seven lipid classes including polar lipid (Pol), free fatty acids (FFA), sterol (ST), hydrocarbon and wax (HW), triacylglycerol (TG), diacylglycerol (DG), and monoalkydiacylglycerol (MADG). Using the GC-FID technique, we have identified 29 fatty acids classified into 3 groups of saturated fatty acid, monounsaturated fatty acids, polyunsaturated fatty acids with an average content of 44.93%, 24.57% and 27.44%, respectively. Among those, many value fatty acids have been detected with high content such as C18:3n-3, C20:4n-6, 20:5n-3, and 22:6n-3. The lipid of 13 brown seaweed Sargassum species also fully contains omega-3,6,9 fatty acids with the content of 9.28%, 16.28% and 16.63%, respectively. Keywords: Omega-3,6,9, Sargassum, Con Dao. Citation: Thu Hue Pham, Van Tuyen Anh Nguyen, Thi Hoang Yen Kieu, Le My Anh Nguyen, Hai Nam Hoang, Quoc Long Pham, Duc Tien Dam, Thi Thanh Van Tran, Tat Thanh Le, 2021. Content and composition of lipid classes, fatty acid from Sargassum seaweed collected at Con Dao and Van Phong bay. Vietnam Journal of Marine Science and Technology, 21(2), 311–318. 311
2. Thu Hue Pham et al. INTRODUCTION organic phase was separated on the funnel, and The genus Sargassum (Phaeophyta) species the total lipid was obtained after evaporation. are found primarily in tropical and subtropical Lipid classes analysis shallow marine waters [1]. Due to their high Thin-layer chromatography (TLC) for the concentration in polysaccharides, natural main lipid classes was carried out on the 6 cm x richness in minerals, polyunsaturated fatty 6 cm plates covered with silica gel (Sorbfil, acids, and vitamins with their bioactivities are Krasnodar, Russia). The mobile phase was n- known to be a good source of healthy food [2]. In Vietnam, there are more than 1,000 hexane/di-ethyl ether/acetic acid 85:25:5 seaweed species, in which 143 species belong (v/v/v). Each sample was tested in triplicate on to the Phaeophyta phylum, including 35 TLC. Then the 10% H2SO4/MeOH reagent was sprayed to cover plates. Then the layer was species of Sargassum genus with 22 species in o the North and 13 species in the South [3]. heated at 210 C for 20 mins. Lipid classes were Their bioactive compounds that are most identified by comparison with standards. The extensively studied include sulfated chromatograms were scanned by Epson polysaccharides, phlorotannins, and diterpenes. Perfection 2400 PHOTO (Nagano, Japan) in a These compounds have been reported to grayscale mode with standard conditions. The possess anti-viral, anti-tumor, and anti-cancer content of the lipid class was determined based properties [2]. However, studies on lipids and on the area and color intensity using the image fatty acids on Sargassum species are rarely analysis program Sorbfil TLC conducted in Vietnam. Videodensitometer, Krasnodar, Russia [8]. The lipid content and fatty acid Fatty acids analysis composition of seaweed are related to FAMEs were prepared following the environmental factors, the difference between ISO/DIS 5509:1998, Germany [9]. The lipid species, and collected place [4–6]. This study was treated with the mixture of 2% H2SO4 in investigates and compares the fatty acid o CH3OH and incubated at 80 C for 2 hours. composition of thirteen seaweed samples Then the mixture of 2 mL n-hexane and 1 mL belonging to the genus Sargassum from Con H2O was added and vortexed for 3 minutes at Dao and Van Phong Bay, Vietnam. room temperature. After that, the hexane phase containing FAMEs was recovered, and the MATERIALS AND METHOD crude FAMEs was purified by TLC using Material hexane/diethyl ether 95:5 (v/v) as mobile phase Thirteen seaweeds belong to the Sargassum and analyzed by gas chromatography Shimadzu genus were collected from Con Dao and Van GC-2010 (Kyoto, Japan). This instrument was Phong, Vietnam, and identified by Dr. Dam equipped with a flame ionization detector (FID) Duc Tien in 2017 (table 1). The collected on an Equity 5 capillary GC column (Merck, samples were cleaned and stored under o length ì internal diameter = 30 m ì 0.25 mm, standard conditions at -20 C. film thickness = 0.25 àm). Methods Lipid analysis RESULTS The total lipid was extracted according to Total lipid the modified method of Bligh and Dyer (1959) The total lipid contents of 13 seaweed [7]. 100 g seaweed was chopped down, samples are ranged from 0.10–1.70% of the fresh extracted with 200 mL MeOH and 100 mL weight (table 1). In detail, only 29KT sample has CHCl3, then put into the ultrasonic bath for 60 a total lipid content of more than 1%, while minutes, at 37 kHz. Then, solid-phase was others have those less than 1%. This result is subjected for the second extraction using similar with previous reports such as from 0.11– 100 mL CHCl3 and ultrasonic assisting for 0.99% in Vietnamese brown seaweeds [10], from 1 hour. The two extractions were combined, 0.1–0.6% in Vietnamese red seaweeds [11, 12], then added 80 mL H2O, and shaken; the lipid from 0.6–1.0% in Indian seaweeds [13]. 312
3. Content and composition of lipid classes Table 1. Total lipid content (% of fresh weight) Name of species Lable Place Time Total lipid (% FW) Sargassum binderi Sond 6KT Con Dao 07/2017 0.90 Sargassum echinocarpum J. Ag. 9KT Con Dao 07/2017 0.70 Sargassum kuetzing Setchell 10KT Con Dao 07/2017 0.32 Sargassum polycystum C. A 20KT Con Dao 07/2017 0.43 Sargassum duplicatum J.Ag. 25KT Con Dao 07/2017 0.31 Sargassum heterocystum Mont 26KT Con Dao 07/2017 0.41 Sargassum mcclurei Seychell 28KT Con Dao 07/2017 0.48 Sargassum paniculatum J. Ag. 29KT Con Dao 07/2017 1.70 Sargassum feldmanii Pham-Hoangho 30KT Con Dao 07/2017 0.10 Sargassum angustyfolium Grunow 6B Van Phong 08/2017 0.62 Sargassum incassum Grunow 7B Van Phong 08/2017 0.24 Sargassum longifructum Tseng et Lu 8B Van Phong 08/2017 0.27 Sargassum crassifolium J. Ag. 11B Van Phong 08/2017 0.40 Lipid class content and composition respectively. The HW and ST classes have an Seven lipid classes have been determined in average content of 11.14% and 10.27%. Finally, 13 species belonging to the Sargassum genus the DG and MADG classes showed in 4 or 5 (table 2), in which five classes, including Pol, samples with a low content of 0.97% and 3.69%, FFA, ST, TG, HW, are found in all species, respectively. The content of the unknown class while diacylglycerol (DG) and was 2.79% and appeared in 4 samples. This monoalkydiacylglycerol (MADG) have appeared result has demonstrated that the number of lipid in several species. In particular, the FFA, TG, classes from the Sargassum species in this and Pol classes possess high content with an research is higher than those in other Sargassum average of 25.52%, 22.95% and 22.42%, species reported by Pham et al., [10]. Table 2. The content and composition of lipid classes Content and composition (% total lipid) Samples Pol ST DG FFA TG MADG Unknow HW 6KT 22.8 11.6 nd 16.0 31.6 11.5 nd 6.5 9KT 16.2 9.9 nd 11.0 41.0 12.7 nd 9.4 10KT 33.3 14.5 nd 7.2 31.2 9.1 nd 4.7 20KT 29.2 9.0 nd 38.1 14.1 nd 9.2 0.5 25KT 17.6 11.2 2.2 37.5 16.0 nd 14.4 1.1 26KT 23.6 10.9 2.0 27.0 18.9 nd nd 17.6 28KT 15.0 9.0 nd 34.2 19.8 nd 6.9 15.1 29KT 16.4 7.5 nd 40.8 15.5 nd 5.8 14.0 30KT 19.6 10.2 2.2 27.4 20.4 nd nd 20.2 6B 22.9 10.8 1.9 27.7 16.7 nd nd 20.1 7B 22.0 9.0 nd 28.6 21.8 nd nd 18.6 8B 33.5 15.6 nd 36.3 7.5 nd nd 7.1 11B 19.4 4.3 4.3 - 43.8 14.7 nd 9.9 Average 22.42 10.27 0.97 25.52 22.95 3.69 2.79 11.14 Notes: nd: Not detected, Pol: Polar lipids, ST: Sterol, DG: Diacyglycerol, FFA: Free fatty acids, TG: Triacyglycerol, MADG: Monoalkydiacylglycerol, HW: Hydrocarbons and wax. 313
4. Thu Hue Pham et al. The fatty acid content and composition 10.00% (9KT) to 35.92% (25KT), with the 6B There are 29 fatty acids identified from 14 sample having the lowest content of 3.4%. to 22 carbon atoms (C14 to C22), in which six The studied samples have remarkedly high popular FAs in seaweeds are C14:0, C16:0, PUFAs content compared to the Northern C18:0, C18:1n-7, C18:1n-9 and C18:3n-3 Sargassum species from 1.28–23.97% [10], appeared in 13 samples. The fatty acids the green seaweeds from 14.8–29.2% [15] and composition of 13 Sargassum species is more from 12.2–39.0% [16]. The benefits of PUFA diverse than that of Sargassum species are well documented for preventing of several collected from Northern Vietnam with 17 fatty types of cancer and various biological acids [10] but less than that of the Hypnea and activities [17]. It has been confirmed that Gracilaria genus in Vietnam with 53 fatty seaweeds contain higher levels of PUFAs than acids found in seven Hypnea samples and 56 vegetables [18]. fatty acids in 48 Gracilaria samples [11]. Among PUFAs, the C20 long-chain fatty acids, including C20:3n-3, C20:4n-6 Saturated fatty acids (SFA) (Arachidonic acid - AA), and C20:5n-3 In the Sargassum genus, the saturated fatty (Eicosapentaenoic acid - EPA) are the most acid (SFAs) content is averaged at 44.93% considered because those fatty acids play (%TL), reaches the highest point at 73.63% critical roles in the immune and inflammatory (9KT), and the lowest point at 19.62% (20KT). responses [19]. Numerous bioactivate lipid C16:0 is the most popular saturated fatty acid mediators have been derived from 20-carbon in the seaweed with the high content reached that produced prostaglandins in the the highest point at 62.12% in the 9KT sample inflammation [17, 20]. Notably, AA has a high and the lowest at 14.96% in the 20KT. content in 10/13 samples (> 10%) and reaches Although the C19:0 fatty acid is rare to be 27.12% in the 20KT sample. Additionally, the identified in seaweeds, it is found in the 6B super long carbon-chain C22:6n-3 (DHA) fatty sample with a content of 0.98%. The fatty acid acid is found in 7 samples with the content contents of the Sargassum seaweeds in this ranged from 0.15–1.05%. study are lower than those of the Sargasssum Fatty acids of the omega-3 group species collected from Thua Thien Hue and All samples contain omega-3 fatty acids Quang Ninh coast reported by Pham et al., [10]. with an average content of 9.28% ranging from Monounsaturated fatty acids (MUFA) 2.9–19.39%. Table 2 showed the omega-3 The MUFA content is from 15.50–28.24%. highest content at 19.39% (20KT) and the C18:1n-9 and C18:1n-7 have been identified in lowest content of 2.9% in 9KT. There are all species with the average content of 15.67% several important fatty acids including 18:3n-3, and 1.67%, respectively, which are important 18:4n-3, C20:3n-3, C20:4n-3, C20:5n-3 (EPA), and C22:6n-3 (DHA). fatty acids belonging to the MUFA group relating to the phytochemicals in plants [11]. Fatty acids of the omega-6 group Interestingly, a very long-chain fatty acid has The omega-6 fatty acids have been been found with the formula of C22:1n-9, observed in all samples with an average content which is rare to appear in seaweeds [10, 14]. of 16.28% varying from 0.43–28.67%. The highest content has shown in 29KT (28.79%), Polyunsaturated fatty acids (PUFAs) followed by 20KT (28.67%), rich in AA The PUFA content is very different among C20:4n-6 with 24.70% and 27.12%, species, ranging from 3.4–53.18% with an respectively. Arcording to a previous study, average of 27.44%. The 20KT and 26KT Pham et al., (2017) has reported that both AA samples, having PUFAs content at 53.18% are very poor in Sargassum species harvested and 40.26%, respectively, are promised for from Co To (Quang Ninh, Vietnam) and Lang applying in the food and the medicine. Ten Co (Thua Thien Hue, Vietnam), with the samples have PUFAs xcontents varying from content ranging from 1.56% to 13.26% [10]. 314
5. Content and composition of lipid classes Table 3. Fatty acid content and composition of samples (% of total fatty acid) Samples 6KT 9KT 10KT 20KT 25KT 26KT 28KT 29KT 30KT 6B 7B 8B 11B 12:0 nd nd 0.30 nd nd nd nd nd nd 1.36 nd nd nd 14:0 10.10 3.88 5.63 3.37 5.34 4.21 3.72 2.86 4.84 7.53 6.87 6.67 3.81 15:0 nd 1.03 0.48 0.22 0.40 0.39 0.62 0.43 0.52 2.47 0.49 0.51 0.45 16:0 40.98 67.12 30.32 14.96 30.33 24.61 30.27 35.62 37.48 52.63 34.64 36.81 42.63 17:0 0.18 0.18 0.35 0.24 0.24 0.24 0.24 nd nd 0.49 0.32 0.29 nd 18:0 1.02 1.29 0.66 0.17 0.52 0.64 2.18 1.07 1.37 1.76 0.81 0.99 1.14 19:0 nd nd nd nd nd nd nd nd nd 0.98 nd nd nd 20:0 2.16 nd 0.21 0.52 0.74 0.55 0.94 0.17 1.14 nd 0.10 0.78 0.60 22:0 0.32 0.15 0.13 0.14 0.12 nd 0.34 0.27 ND 0.51 nd nd nd ΣSFA 54.75 73.63 38.09 19.62 37.67 30.64 38.31 40.41 45.35 67.71 43.23 46.05 48.64 16:1n-7 4.96 2.82 10.26 6.88 7.18 7.55 6.68 6.47 5.40 nd 7.99 6.46 5.15 16:1n-5 1.86 nd 0.23 0.64 0.37 0.47 0.44 0.46 0.98 4.78 0.58 0.19 0.46 18:1n-11 nd nd nd nd nd nd nd 5.29 nd nd nd nd nd 18:1n-9 6.46 10.58 18.16 16.79 16.56 17.80 19.67 11.42 16.88 16.36 18.99 16.72 17.37 18:1n-7 6.21 2.09 1.30 0.31 0.65 1.19 1.23 1.25 0.71 4.40 0.66 1.27 0.48 22:1n-9 nd nd nd nd nd nd 0.22 nd nd nd nd nd nd ΣMUFA 19.04 15.05 29.96 24.62 24.76 27.01 28.24 24.88 23.97 25.55 28.22 24.64 23.46 18:2n-6 0.31 0.10 1.65 0.21 0.12 3.01 nd 1.69 2.28 nd 1.27 1.61 2.68 20:2n-6 0.36 0.11 0.81 0.19 0.17 0.21 0.68 0.48 0.68 nd 0.11 0.12 nd 315
6. Thu Hue Pham et al. 22:2n-6 nd 4.07 nd nd nd nd nd nd nd nd nd nd nd 18:3n-6 0.41 0.11 0.55 0.95 0.70 0.42 0.53 0.65 nd nd 0.47 0.49 0.62 18:3n-3 4.28 1.44 7.77 5.34 5.57 6.09 6.06 0.16 3.80 1.13 6.35 6.58 5.14 20:3n-9 0.54 0.24 1.07 2.19 1.47 1.54 0.79 1.27 0.84 nd 0.64 0.64 0.97 20:3n-6 0.25 nd 0.75 0.21 0.92 1.20 0.99 1.27 0.72 0.43 0.49 0.50 0.97 20:3n-3 0.45 nd nd 0.25 nd nd 0.52 0.53 nd 1.83 nd nd nd 18:4n-3 nd nd 0.26 7.71 4.78 0.25 2.46 0.77 0.33 nd 0.60 0.36 0.59 20:4n-3 0.48 0.21 0.29 2.93 1.52 1.85 0.25 nd 1.11 nd 1.05 1.41 1.12 20:4n-6 3.64 2.26 15.86 27.12 15.34 19.96 12.77 24.70 12.83 nd 12.91 13.95 12.83 20:5n-3 2.46 1.31 1.92 6.09 5.17 4.69 2.65 3.19 2.10 nd 1.40 1.80 2.56 22:5n-3 0.53 nd nd nd nd nd 0.54 nd 0.31 nd Nd nd nd 22:6n-3 nd 0.15 0.11 nd 0.16 1.05 0.61 nd nd nd 0.14 0.30 nd Other 12.03 0.86 0.91 2.58 1.64 2.10 4.58 0.00 5.68 3.34 3.12 1.55 0.41 ΣPUFA 13.72 10.00 31.05 53.18 35.92 40.26 28.86 34.71 24.99 3.40 25.42 27.76 27.49 Omega-3 7.73 2.90 10.07 19.39 15.69 12.07 12.85 4.65 6.54 2.96 8.49 9.04 8.30 Omega-6 4.97 6.66 19.62 28.67 17.25 24.80 14.97 28.79 16.50 0.43 15.24 16.67 17.10 Omega-9 7.00 10.82 19.23 18.98 18.04 19.34 20.68 12.69 17.73 16.36 19.63 17.36 18.33 PUFA/SFA 0.25 0.14 0.82 2.71 0.95 1.31 0.75 0.86 0.55 0.05 0.59 0.60 0.57 n-3/n-6 1.56 0.44 0.51 0.68 0.91 0.49 0.86 0.16 0.40 6.84 0.56 0.54 0.49 Notes: nd: not detected, SFA: Saturated Fatty Acids, MUFA: Monounsaturated Fatty Acids, PUFA: Polyunsaturated Fatty Acids. 316
7. Content and composition of lipid classes Fatty acids of the omega-9 group 1.70% of the fresh weight. The PUFA/SFA and The omega-9 fatty acids have been found in n-3/n-6 index are suitable with the WHO all samples with an average value of 16.63%. recommendation. Thus lipids of these species The 28KT sample has the highest content of belonging to the Sargassum genus are good for 20.68%, while the 6KT has 7.00%. These health. Five lipid classes, including Pol, ST, results are similar to a previous publication [10, FFA, TG, HW, have been identified in almost 11]. The main fatty acid is 18:1n-9, with the all species, while two lipid classes, including content ranged from 6.46% to 19.67%. DG and MADG, are present in 4 to 5 samples. Twenty-nine fatty acids have been detected, in PUFA/SFA index which SFA has an average content of 44.93%; In table 2, 10/13 samples (excepted 6KT, the MUFA has an average content of 24.57%; 9KT and 6B) have the PUFA/SFA index being PUFA has a moderate content of 27.44%. higher than 0.4, which is recommended good Among unsaturated fatty acids, various for health by The World Health Organization valuable fatty acids belong to omega-3,6,9, (WHO). In particular, two samples including such as C18:3n-3, C20:4n-6, 20:5n-3, and 20KT and 26KT have the high index of 2.71 22:6n-3. and 1.31, while others are between 0.55 (30KT) and 0.95 (25KT). REFERENCES n3/n6 index [1] De Wreede, R. E., 1976. The phenology All species have the n-3/n-6 index ranging of three species of Sargassum from 0.16 (29KT) to 6.88 (6B). The WHO (Sargassaceae, Phaeophyta) in Hawaii. currently recommended the n-3/n-6 ratio to be Phycologia, 15(2), 175–183. doi: higher than 0.1 in diet to prevent 10.2216/i0031-8884-15-2-175.1 inflammation, cardiovascular and nervous [2] Gupta, S., and Abu-Ghannam, N., 2011. system disorders [21]. Bioactive potential and possible health From the results of the total lipid and fatty effects of edible brown seaweeds. Trends acids of 13 brown seaweed Sargassum species, in Food Science & Technology, 22(6), we have reconigzed that three species, 315–326. including S. polycystim (20KT), S. heterocyst 2011.03.011 (26KT), and S. paniculatum (29KT), have a [3] high value on the lipid composition. In my/?tc=accept&id=4360; accessed July particular, both of them belong to the 3rd top of 30, 2020. the PUFA content, omega-6 content, and [4] Chen, Z., Xu, Y., Liu, T., Zhang, L., Liu, PUFA/SFA index. The PUFA content of the H., and Guan, H., 2016. Comparative three samples is 4.6 to 7.1 times higher than studies on the characteristic fatty acid those of the species collected from Quang Ninh profiles of four different Chinese to Thua Thien Hue, Vietname [10]. Also, three medicinal Sargassum seaweeds by GC- samples contain AA and EPA with high content MS and chemometrics. Marine drugs, ranging from 19.96–27.12% and from 3.19– 14(4), 68. 6.09%, respectively. Thus, they could be merit md14040068 to further study on the chemicals and [5] Gerasimenko, N., and Logvinov, S., 2016. bioactivities for the application on the food and Seasonal composition of lipids, fatty acids the medicine. pigments in the brown alga Sargassum pallidum: The potential for health. Open CONCLUSIONS Journal of Marine Science, 6(04), 498– In summary, we have determined the total 523. doi: 10.4236/ojms.2016.64041 lipid and content and composition of the lipid [6] Cohen, Z., Vonshak, A., and Richmond, class of 13 species collected from Con Dao and A., 1988. Effect of environmental Van Phong in Vietnam. The total lipid content conditions on fatty acid composition of of Sargassum seaweeds is ranged from 0.1– the red alga Porphyridium cruentum: 317
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