Nutritional Components and Antioxidant Activities of Sea Buckthron (Hippophae rhamnoides L.) Leaf and Berry Extracts

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1 J East Asian Soc Diet Life 28(: (2018) , 2 Nutritional Components and Antioxidant Activities of Sea Buckthron (Hippophae rhamnoides L.) Leaf and Berry Extracts So-Young Lee 1, Jong-Yun Shon 2 and Kun-Og Kang 1 1 Dept. of Nutrition and Culinary Science, Hankyong National University, Ansung 17579, Korea 2 Dept. of Food Biotechnology & Institute of Food Industry and Biotechnology, Hankyong National University, Ansung 17579, Korea ABSTRACT Nutritional components and antioxidant activities of sea buckthron (Hippophae rhamnoides L.) leaf and berry extracts were evaluated. The fructose contents of leaves (6.64 g/100 g) and berries (2.92 g/100 g) were higher than those of other free sugars, such as glucose, and sucrose. Organic acids (malic acid, lactic acid, acetic acid) were detected, with malic acid ( mg/g) being most abundant. Nine fatty acids were detected, with linolenic acid (42.38%) being most abundant. The total phenol contents of leaves extracted with water, 50% ethanol, and 80% ethanol were mg/g, mg/g, and mg/g, respectively. Therefore, extractions with ethanol were more effective than those with water. Moreover, the total flavonoid content of leaves (32.39 mg/g) extracted by 50% ethanol was higher than that of the other extracts. The order of nitrite-scavenging abilities at ph 1.2 was ascorbic acid (92.62%)>50% ethanol (93.07%)>80% ethanol (90.66%)>water (84.34%). Electron donating ability was highest for leaves (87.62 mg/ml) extracted with 50% ethanol. Overall, the results of this study suggest that sea buckthron has nutritional effects and antioxidant activities, making it a great potential functional food. Key words : Sea buckthron, nutritional components, antioxidant activity,,., (Oh KJ 2015; Kim SH & Jeong YJ 2012). (Hippophae rhamnoides L.),. C, E,, (Tiffany TYG 2005). Corresponding author : Kun Og Kang, Tel: , Fax: , cocco-9522@hanmail.net,, (Park YH 2010a), (Park YH 2010b), (Ko MS 2012), (Kim EJ 201, (Kim MW 2013), (Choi SJ 2016).,, 50% 80%. 1. (,

2 ) (FD-8515, Ilsin, Korea) (M20, IKA, Germany) mesh, AOAC (A.O.A.C. 1995)., 105, Soxhlet, semi micro Kjeldahl (N 6.25), %,,,. 2) Lee JW (2014) 5 g 25 ml, acetonitrile 50 ml sonicator (Bransonic, 8510E-DTH, CT, USA) ,000 rpm (Combi 514R, Hanil, Gangwon-do, Korea) 15, 0.45 μm membrane filter. fructose, glucose, sucrose, lactose, maltose(sigma-aldrich Chemical Co. St. Louis, MO, USA) 0.2 mg/g, 0.5 mg/g, 1 mg/g, 2 mg/g, 5 mg/g, 10 mg/g, Table 1 HPLC. 3) Bang CM (2016). 1 g 50 ml (3,000 rpm, 15 min), 0.45 μm membrane filter. Column KC-LG KC-811 Shodex RS pak KC-LG guard. citric acid, tartaric acid, malic acid, succinic acid, lactic acid, acetic acid (Sigma-Aldrich Chemical Co. St. Louis, MO, USA) 0.2 mg/g, 0.4 mg/g, 0.6 mg/g, 0.8 mg/g 1 mg/g, Table 2 HPLC. 4) AOCS (A.O.C.S. 1990)., 250 ml g 0.5 N NaOH/methanol 4 ml, % BF 3 /methanol 5 ml 2 hexane 5 ml 1., test tube, hexane Table 3. 5) ( (MCH600SI, Tongyang Magic Co., LTD., Seoul, Korea) 40 mesh 20 g 20, 50% 80% (Whatman No. 2), 3. 40±1 (rotary Table 2. HPLC analysis conditions for organic acid Table 1. HPLC analysis conditions for free sugar Items Instrument Waters 2690 Detector Column Column temp. Solvent Conditions Waters 410(Differential refractometer) Shodex Asahipak NH 2P 50 4E(250 mm 4.6 mm ID) 35 CH 3 CN/H 2 O=75/25 Items Instrument Detector Column Column temp. Solvent Conditions Shiseido nanospace SI-2 UV/VIS detector 440 nm(shiseido nanospace si ) Shodex RS pak KC-LG(8.0 mm 50 mm ID) +KC-811(8.0 mm 300 mm ID) 2 70 A solvent: 3 mm perchloric acid B solvent: 0.2 mm bromothymolblue, 15 mm Na 2 HPO 4, 2 mm NaOH Flow rate 1.0 ml/min Flow rate A: 0.7 ml/min, B: 0.7 ml/min Injection volume 20 ul Injection volume 10 μl

3 28(: (2018) 33 Items Table 3. GC analysis conditions for of fatty acid Instrument Column Carrier gas Injection port Detection port Oven temp. program Conditions Youngin M600D Supelco 2560(100 m 0.25 mm 0.2 μm) He(1.0 ml/min) Initial temp. 100 at 4 min Temp. gradient 3 /min Final temp. 240 at 15 min evaporator N-1000, Eyela, Tokyo, Japan) (freeze dryer, FD, DF8517, ilshinbiobase Co Ltd, Dongducheon, Korea). 40 (MDF- U50V, Sanyo, Tokyo, Japan). (2) Folin O & Denis W(1912) (500 μg/ml) 0.2 ml 2% sodium carbonate 2 ml 3. 1 N Folin-Ciocalteu's phenol reagent(sigma-aldrich Chemical Co. St. Louis, MO, USA) 0.1 ml nm. gallic acid(sigma-aldrich Chemical Co.), gallic acid 0 μg/ml, 50 μg/ ml, 150 μg/ml 200 μg/ml. g mg gallic acid equivalent. (3) (Kang YH 1996) (2 mg/ml) 1 ml diethylene glycol 10 ml, 1 N NaOH 1 ml 37 1, 420 nm. quercetin(sigma-aldrich Chemical Co.), quercetin 0 μg/ml, 25 μg/ml, 50 μg/ml, 75 μg/ml 100 μg/ml. g mg quercetin. (4) (NaNO 2 ) Gray JI & Dugan LR(1975)., 1 mm NaNO 2 2 ml (2 mg/ml) 1 ml 0.1 N HCl(pH 1.2), 0.2 M (ph 3.0 ph 6.0) ph 1.2, ml ml 2% 5 ml Griess (30% 1% sulfanilic acid 1% naphthylamine 1:1 ) 0.4 ml nm 3. Griess 0.4 ml., ascorbic acid(2 mg/ml). N(%) = 1 A C 100 B N: A: 1 mm NaNO 2 1 B: 1 mm NaNO 2 1 C: (5) DPPH DPPH (electron donating ability; EDA) Blois MS(1958)., ( mg/ml) 1.0 ml, M DPPH(1,1-diphenyl-2-picrylhydrazyl; Sigma Co.) 4.0 ml nm 3. DPPH (%), DPPH 50% (mg/ml) IC 50. ascorbic acid(sigma-aldrich Chemical Co.) (0.01 mg/ml, 0.02 mg/ml, 0.03 mg/ml 0.04 mg/ml). EDA(%) = (1 A/B) 100 A: B: 3. SAS package(release 8.01, SAS Institute., Cary, NC, USA) ±,

4 34 p<0.05 Duncan's multiple range test. 1. Table %, 10.49%, 17.15%, 10.48%, 6.12%, 12.27%, 3.99%, 2.77% 61.15%, 63.99%.. 2. Table 5 fructose, glucose, sucrose, sucrose, 9.15 g/100 g, 4.78 g/100 g 2. fructose 6.64 g/100 g, sucrose 1.54 g/100 g, glucose 0.97 g/100 g, fructose 2.92 g/100 g, glucose 1.86 g/100 g. Kim JH (2003) fructose, sucrose, glucose 7.31 g/100 g, 2.48 g/100 g, 0.08 g/100 g. Jo AK(2015) fructose, glucose, sucrose 1.14 g/100 g, 0.73 g/100 g, 0.55 g/100 g,. Shin SR(1999) fructose, glucose, sucrose 3, sucrose Table 4. Approximate compositions of sea buckthron (Hippophae rhamnoides L.) leaf and berry (dry base %) Leaf Berry t Moisture 11.59±0.17 a 10.49±0.01 b *** Crude protein 17.15±0.74 a 10.48±0.18 b *** Crude lipid 6.12±0.16 b 12.27±0.16 a 2, *** Crude ash 3.99±0.03 a 2.77±0.15 b *** Carbohydrate 61.15±0.28 b 63.99±0.13 a *** a,b Values with different superscript within a same row are significantly different(p<0.05) by Duncan's multiple test. *** p< Table 5. Contents of free sugar in sea buckthron (Hippo phae rhamnoides L.) leaf and berry (g/100 g) Free sugar Leaf Berry t Fructose 6.64±0.09 a 2.92±0.61 b *** Glucose 0.97±0.03 b 1.86±0.01 a 2, *** Sucrose 1.54± Total 9.15±0.13 a 4.78±0.62 b *** a,b Values with different superscript within a same row are significantly different(p<0.05) by Duncan's multiple test. *** p< g/100 g, 1.38 g/100 g, Jung GT (2013) sucrose glucose fructose,. 3. Table 6 3, mg/g mg/g 8. malic acid mg/g. Acetic acid 6.76 mg/g 6.19 mg/g lactic acid (6.50 mg/g) (4.89 mg/g).,,, (Jeong CH 2005) oxalic acid, tartaric acid, malic acid, citric acid, succinic acid 5, succinic acid mg/g, (Jung GT 2013) malic acid succinic acid. 4. Table 7, Fig. 1 8, 6., palmitic acid (C16:0) stearic acid(c18:0) 2, mysristic acid(14:0), arachidic acid(20:0), behenic acid(22:0). oleic acid(18:, linoleic acid(18:2), linolenic acid(18:3) 3, palmitoleic acid(16:

5 28(: (2018) 35 Table 6. Contents of organic acid in sea buckthron leaf and berry (mg/g) Organic acid Leaf Berry t Malic acid 2.83±0.03 b ±0.29 a 4, *** Lactic acid 6.50±0.02 a 4.89±0.07 b 1, *** Acetic acid 6.19±0.04 b 6.76±0.09 a *** Total 15.52±0.09 b ±0.45 a 1, *** a,b Values with different superscript within a same row are significantly different(p<0.05) by Duncan's multiple test. *** p< linolenic acid 42.38%, behenic acid 11.93%, palmitic acid 11.51%, linoleic acid 10.35%. palmitic acid 30.44%, palmitoleic acid 28.44%, linoleic acid 15.86%, stearic acid 11.26%, oleic acid 8.74% %, 58.28%, 31.21%, 41.70%. Jeong CH (2005) 9, linoleic acid 21.15%, oleic acid 20.92%, myristic acid 20.18%, palmitic acid 17.59%, linolenic acid 14.28%. Jo AK(2015),, oleic acid, linoleic acid, linolenic acid 18.88%, 40.79%, 17.78%, Table 7. Fatty acid composition of sea buckthron leaf and berry (%) Fatty acid Leaf Berry t Myristic acid(14:0) 2.07± Palmitic acid(16:0) 11.51±0.11 b 30.44±0.16 a *** Palmitoleic acid(16: ± Stearic acid(18:0) 2.72±0.04 b 11.26±0.08 a *** Oleic acid(18: 6.13±0.05 b 8.74±0.06 a *** Linoleic acid(18:2) 10.35±0.09 b 15.86±0.08 a *** Linolenic acid(18:3) 42.38±0.17 a 5.24±0.03 b 1, *** Arachidic acid(20:0) 2.98± Behenic acid(22:0) 11.93± ΣSFA 31.21±0.18 b 41.70±0.24 a *** ΣUSFA 2) 58.86±0.31 a 58.28±0.29 a 5.60 * Sum of saturated fatty acids. 2) Sum of unsaturated fatty acids. a,b Values with different superscript within a same row are significantly different(p<0.05) by Duncan's multiple test. * p<0.05, *** p< linolenic acid. 5. phytochemical Leaf Berry Fig. 1. Fatty acid composition of sea buckthron leaf and berry.

6 36, 2. (Jeong CH, 201. Fig. 2 80% ( mg/g)>50% ( mg/g)> ( mg/g) 80%. 50% (68.11 mg/g)>80% (62.87 mg/g)> (56.59 mg/g), 2.5.,. Kim JH & Kang KO(2016) 47.9 mg/g, 53.4 mg/g, Lim JH(2009) 58.5 mg/g, mg/g. 2) Fig. 3 50% (32.39 mg/g)>80% (26.07 mg/g)> (24.04 mg/g) 50%. 50% (5.87 mg/g)>80% (5.50 mg/g)> (4.48 mg/g), Fig. 3. Total flavonoid contents of sea buckthron leaf and berry extracts. a c Means with the different letters above the bars are significantly different(p<0.05) by Duncan s multiple test %, 9.4% (Kim SG 2012), (Kang KO 2013) (9.2%) (7.8%),. Fig. 2. Total phenol contents of sea buckthron leaf and berry extracts. a e Means with the different letters above the bars are significantly different(p<0.05) by Duncan s multiple test. 3) nitroso nitrosamine (Kyun SH 2007). amine nitrosamine., 50%, 80% ascorbic acid ph 1.2, ph 3.0 ph 6.0 Table 8. ph ph,. ph % (93.97%)>ascorbic acid(92.62%)>80% (90.66%)> (84.34%) 50%

7 28(: (2018) 37 Table 8. Nitrite-scavenging ability of sea buckthron leaf, berry extracts and ascorbic acid Leaf extracts Berry extracts Sample ph condition ph 1.2 ph 3.0 ph 6.0 F Water 84.34±3.32 ab 26.93±3.56 bd 9.10±0.07 cd *** 50% EtOH 93.07±4.56 aa 75.41±6.71 ba 11.13±0.23 cc *** 80% EtOH 90.66±3.12 aa 80.39±6.41 ba 13.15±0.02 cb *** F 4.40 * ** *** - Water 11.60±0.07 ad 7.46±0.78 be 3.18±0.56 cf *** 50% EtOH 73.04±4.41 ac 67.68±3.46 ab 10.55±0.78 cc *** 80% EtOH 71.99±2.31 ac 67.13±0.03 bb 7.23±0.98 ce 1, *** Ascorbic acid 92.62±0.94 aa 53.03±0.63 bc 19.61±1.78 ca 2, *** F *** *** *** - a c Values with different superscript within a same row are significantly different(p<0.05) by Duncan s multiple test. A E Values with different superscript within a same column are significantly different(p<0.05) by Duncan s multiple test. * p<0.05, ** p<0.01, *** p< ascorbic acid. 50% (73.04%)>80% (71.99%)> (11.60%), 50% ph 1.2. Lee SJ (2000),, ph %, ph %, ph % ph,. Ju MJ (2009) ph, Park CS (2002) ph %, 27%. 4) DPPH DPPH free radical radical (Blois MS, 1958). DPPH, free radical (Ko MS 2012). Fig. 4 50% (87.62%)>80% (83.12%)> (77.33%) ascorbic acid 91.11%. 50% (28.47%)> (22.73%)>80% (19.63%),. Ko MS (2012) Fig. 4. DPPH radical scavenging activity of sea buckthron leaf, berry(0.1 mg/ml) extracts and ascorbic acid(0.04 mg/ml). a e Means with the different letters above the bars are significantly different(p<0.05) by Duncan s multiple test.

8 38, ascorbic acid, Choi SJ (2016), g/100 g, 4.78 g/100 g 2. fructose 6.64 g/100 g, sucrose 1.54 g/100 g, glucose 0.97 g/100 g, fructose 2.92 g/100 g, glucose 1.86 g/100 g. Malic, lactic, acetic acid mg/g mg/g 8, malic acid mg/g. palmitic acid(c16:0) stearic acid(c18:0) 2, oleic acid(18:, linoleic acid(18:2), linolenic acid(18:3) 3. linolenic acid 42.38%, palmitic acid 30.44%, 58.86%, 58.28%, 31.21%, 41.70%. 80% ( mg/g)>50% ( mg/g)> ( mg/g), 50% (68.11 mg/g)>80% (62.87 mg/g)> (56.59 mg/g) % (32.39 mg/g)>80% (26.07 mg/g)> (24.04 mg/g), 50% (5.87 mg/g)>80% (5.50 mg/g)> (4.48 mg/g). ph, ph % (93.97%)>ascorbic acid(92.62%)> 80% (90.66%)> (84.34%) 50% ascorbic acid, 50% (87.62%)>80% (83.12%)> (77.33%).,. REFERENCES AOAC (1995) Official Methods of Analysis. 16th ed. Association of Official Analytical Chemists, Washington, DC, USA. pp AOCS (1990) Official Methods for the Analysis of Fats, Oils and Related Materials. 4th ed. American Oil Chemists' Society, Cd Bang CM, Moon JK, Hong SK (2016) Characteristics of organic acid of Makgeolli by yeast strains type. Korean J Food Cook Sci 32(: Blois MS (1958) Antioxidant determination by the use of a stable free radical. Nature 181(10): Choi SJ, Kim NR, Shin JY, Lee KM, Kyong KY (2016) Evaluation of anti Wrinkle efficacy in sea buckthorn extracts, oil and nanoemulsion containing them. J Korean Soc Cosmetics & Cosmetology 6(2): Folin O, Denis W (1912) On phosphotungstic-phosphomolybdic compounds as color reagents. J Biol Chem 12(2): Gray JI, Dugan Jr LR (1975) Inhibition of N-nitrosamine formation in model food system. J Food Sci 40(4): Jeong CH, Nam EK, Shim KH (2005) Chemical components in different parts of Erigeron annuus. J Korean Soc Food Sci Nutr 34(6): Jeong CH, Kim IH, Shim KH, Bae YI (201 Nutritional components and antioxidant activities of commercial loquat leaf tea. J Agri Life Sci 45(4): Jo AK (2015) The quality characteristics and antioxidant activities of Yakju and liquor containing rose, camellia and cockscomb flower. Ph D. Dissertation. Hankyong Natioanl University, Anseong. pp Jung GT, Ju IO, Choi SR, You DH, Noh JJ (2013) Food nutritional characteristics of fruit of Cudrania tricuspidata in its various maturation stages. Korean J Food Preserv 20(3): Ju MJ, Kwon JH, Kim HK (2009) Physiological activities of mulberry leaf and fruit extracts with different extraction conditions. Korean J Food Preserv 16(4): Kang KO (2013) Analysis of antioxidant effects and antimicrobial activity of extracts from yacon(polymnia sonchifolia) powder. J East Asian Soc Dietary Life 23(3): Kang YH, Park YK, Lee GD (1996) The nitrite scavenging and electron donating ability of phenolic compounds.

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