Prawarun Agr. J. Volume 15 (SUPPL. 1) 2018, Pages 40-47

Research Article

Nutritional Compositions and Antioxidant Activities of Makamong ( xylocarpa (Kurz) Craib), Flower Fence (Caesalpinia pulchrrima (L.) Sw.) and Tamarind (Tamarindus indica L.) Kernels

Sarin Thongthummachat1, Pornpimol Ponkham1, Suchana Wanich1, Butsayamat Rattanadon1, Metta Kengchuwong2 and Kwanyuen Leamsamrong1*

1Department of Chemistry, Faculty of Science and Technology, Rajabhat Maha Sarakham University, Maha Sarakham, 44000, 2Department of Environment, Faculty of Science and Technology, Rajabhat Maha Sarakham University, Maha Sarakham, 44000, Thailand

Abstract

This study was aimed to investigate the nutritional composition and antioxidant activity of three kernels including makamong (Afzelia xylocarpa (Kurz) Craib), flower fence (Caesalpinia pulchrrima (L.), tamarind (Tamarindus indica L.). It was found that the kernel of makamong shown the highest nutritional compositions, mineral contents and vitamin. Furthermore, the kind of makamong consist of values for energy, ash, total fat, B-carotene and vitamin B2 were 439.42±0.72 kcal, 2.49±0.01%, 10.64±0.07%, 92.59±1.17 µg g-1 DW and 0.11±0.01 mg g-1 DW, respectively. The makamong kernel (MK), flower fence kernel (FW) and tamarind kernel (TM) were extracted with 50% ethanol (50:50; water: ethanol), 80% ethanol (20: 80;water: ethanol) and ethanol. The antioxidant activities were evaluated using DPPH (1,1-diphenyl-s-picrylhydrazyl), ABTS+ (2,2’-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) and FRAP (ferric reducing antioxidant power) assay. The total polyphenol and flavonoid contents were analyzed. The results revealed that 50% ethanolic extract of MK showed the highest DPPH radical scavenging activity and total flavonoid contents (IC50; 8.80±0.03 mg ml-1, 37.95±0.38 mg QE g-1), respectively. The results suggest that makamong kernel could be used as a good source for natural antioxidant food and dietary supplement.

Keywords: Makamong, Flower fence, Tamarind, Kernel, and Antioxidant activity

*Corresponding author: E-mail: [email protected] 40 วารสารเกษตรพระวรุณ ปีที่ 15 (ฉบับพิเศษ 1) พฤษภาคม 2561 Volume 15 (SUPPL. 1) May 2018 Prawarun Agr. J. Volume 15 (SUPPL. 1) 2018, Pages 40-47

องค์ประกอบทางโภชนาการและฤทธิ์ต้านอนุมูลอิสระจากเนื้อในเมล็ดมะค่าโมง (Afzelia xylocarpa (Kurz) Craib) หางนกยูง (Caesalpinia pulchrrima (L.) Sw.) และ มะขาม (Tamarindus indica L.)

ศรินทร์ ทองธรรมชาติ1, พรพิมล พลค า1, สุชนา วินิช1, บุษยมาส รัตนดอน1 เมตตา เก่งชูวงศ์2 และ ขวัญยืน เลี่ยมส าโรง*

1สาขาวิชาเคมี คณะวิทยาศาสตร์และเทคโนโลยี มหาวิทยาลัยราชภัฏมหาสารคาม อ าเภอเมออ จั หวัดมหาสารคาม 44000 2สาขาวิชาสิ่ แวดล้อม คณะวิทยาศาสตร์และเทคโนโลยี มหาวิทยาลัยราชภัฏมหาสารคาม อ าเภอเมออ จั หวัดมหาสารคาม 44000

บทคัดย่อ

วัตถุประสงค์ของการศึกษาครั้งนี้ เพื่อศึกษาองค์ประกอบทางโภชนาการ และทดสอบฤทธิ์ต้านอนุมูลอิสระของเนื้อในเมล็ด 3 ชนิดประกอบด้วย มะค่าโมง (Afzelia xylocarpa (Kurz)Craib) หางนกยูง (Caesalpinia pulchrrima (L.) และมะขาม (Tamarindus indica L.) จากการทดลองพบว่า เนื้อในเมล็ดมะค่าโมงมีองค์ประกอบทางโภชนาการ แร่ธาตุ และวิตามิน นอกจากนี้ เนื้อในเมล็ดมะค่าโมงยังประกอบด้วย พลังงานทั้งหมด เถ้า ไขมันทั้งหมด เบต้าแคโรทีน และ วิตามินบี 12 สูงที่สุด เท่ากับ 439.42±0.72 kcal, 2.49±0.01%, 10.64±0.07%, 92.59±1.17 µg g-1 DW and 0.11±0.01 mg g-1 DW ตามล าดับ เมื่อน าเนื้อเมล็ดมะค่าโมง (MK) หางนกยูง (FW) และมะขาม (TM) มาสกัดด้วยตัวท าละลาย 50% เอธานอล (50 : 50 ; น้ า: เอธานอล), 80% เอธานอล(20: 80;น้ า: เอธานอล) และเอธานอล ส าหรับการทดสอบฤทธิ์ต้านอนุมูลอิสระได้ทดสอบด้วยวิธี DPPH(1,1-diphenyl-s-picrylhydrazyl), ABTS+ (2,2’-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid), FRAP (ferric reducing antioxidant power) และวิเคราะห์หาปริมาณทั้งหมดของโพลิฟีนอลและฟลาโวนอยด์ พบว่า สารสกัดด้วย 50% เอธานอลของมะค่าโมง (MK) มีฤทธิ์ต้านอนุมูลอิสระด้วยวิธี DPPH และมีปริมาณทั้งหมดของฟลาโวนอยด์สูงที่สุด -1 -1 (IC50;8.80±0.03 mg ml , 37.95±0.38 mg QE g ) จากผลการทดลองเนื้อในเมล็ดมะค่าโมงนั้นเป็นพืชที่มีคุณค่าสูง ควรค่า น าไปใช้เป็นแหล่งอาหารต้านอนุมูลอิสระจากธรรมชาติ และน าไปต่อยอดในการผลิตอาหารเสริมสุขภาพได้

ค าส าคัญ: มะค่าโมง หางนกยูง มะขาม เนื้อในเมล็ด และ ฤทธิ์ต้านอนุมูลอิสระ

*ผู้เขียนให้ติดต่อ: E-mail: [email protected] ปีที่ 15 (ฉบับพิเศษ 1) พฤษภาคม 2561 วารสารเกษตรพระวรุณ 41 Volume 15 (SUPPL. 1) May 2018 Prawarun Agr. J. Volume 15 (SUPPL. 1) 2018, Pages 40-47

Introduction activities of MK and FK are not previously

reported. Thus, the aim of this study was to Makamong (Afzelia xylocarpa(Kurz) Craib) is determine the nutritional composition (moisture, a tree from Asia. It was cultivated in Thailand, ash, crude protein, crude fat and carbohydrate), , , , and Burmain deciduous mineral, vitamin and antioxidant activities (DPPH, + forests. The seeds are harvested for medicinal ABTS , FRAP, TPC and TFC) of several these purposes. In Thailand, A. xylocarpa (locally known kernels. as makamong) are planted as shading trees due to its wide-ranging branches. Makamong has used the Materials and Methods different part of roadsides and waterways the tree provides a good wind barrier and protection from 1.Materials, chemicals and reagents water-driven soil erosion. Seed: large, shiny black Makamong, flower fences and tamarind or dark brown, seed coat thickness; is a large, seeds were obtained from local market Muang situated at radicle end, yellow when dry. There Kalasin, Kalasin Province, Thailand. Folin-Ciocalteu are 110-160 seeds per kg (Talumatsawa, 2006). reagent was purchased from Fisher Chemical Flower fence (Caesalpinia pulcherrima (L.) Sw.) is (Leicestershire, UK).6-Hydroxy-2,5,7,8- the belonging to the family Caesalpiniaceae tetramethylchromane-2-carboxylic acid (Trolox), which had the wide range of medicinal uses. 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis Vernacularly was known as flower fence. In India, ( 3-ethylben-zothiazoline-6-sulphonicacid) (ABTS) , several parts of flower fence such as leaves, 2,4,6-Tripyridyl-s-Triazine (TPTZ) , quercetin, gallic flowers, plates, barks and seeds have been used in acid, ascorbic acid were purchased from Sigma (St. Indian medicine (Sastri, 1998). The barks are used Louis, MO, USA). Ferrous chloride was obtained as an abortifacient agent was prepared by from Fluka (Germany). Ethanol was purchased macerating coarse powder for 24 hours by leaves from BDH (UK).The chemicals were used of are used as cathartic (Chakraborthy et al., 2009). analytical grade. Tamarind (Tamarindus indicaL.) is a crucial food and common trees in the tropics. Tamarind is 2. The preparation of dried kernels indigenous to tropical Africa but it has been Seeds from three were washed three introduced and naturalized worldwide in over 50 times in tab water and soaked in 12 h. Then, the countries (Mishra and Malhotra, 2009; De Caluwe seeds were cultivated in burning hush soil with et al., 2010; Takawale et al., 2011). Africa on the daily watering for 7 days. Whole seeds were whole does not produce tamarind on a harvested and removed the seed coat to obtain commercial scale, though it is widely used by the the kernel seed. The makamong kernel (MK), local people. The kernel powder of tamarind is flower fence kernel (FK) and tamarind kernel (TK) used various industry such as sizing material in seeds were dried in a hot air oven at 60°C, gently textile, paper, mining, cosmetic, tobacco and jute milled, sealed in aluminium foil bag to protect industries (Takawale et al., 2011). However, the moisture absorption and stored in freezer at -20 °C nutritional composition and the antioxidant until used for chemical composition and

42 วารสารเกษตรพระวรุณ ปีที่ 15 (ฉบับพิเศษ 1) พฤษภาคม 2561 Volume 15 (SUPPL. 1) May 2018 Prawarun Agr. J. Volume 15 (SUPPL. 1) 2018, Pages 40-47 antioxidant analysis. The kernel powders of MK, FK evaporator at 45 °C and stored at -20 °C until and TK were shown in Fig. 1. further study. 2) DPPH radical scavenging activity The DPPH free radical scavenging activity of kernel extract was determined according to the method of Xu (2003). Briefly, 100 µl of different concentrations of kernel extracts were mixed with 900 µl of 0.1 mM DPPH solution stored in dark for 30 min. The absorbance was measured at 515 nm. The method was used trolox and ascorbic acid as

standard solution. IC50 was the concentration of Fig. 1 The kernel powders of makamong (MK), standard or extracts solution with 50% DPPH flower fence (FK) and tamarind (TK) radical scavenging activity. The radical scavenging

activity was calculated using following equation. 3. Nutritional composition, mineral and vitamin

Proximate analysis of kernels was % determined using the Official Method of Association of Official Analysis Chemical (AOAC, 1995) including moisture, ash, crude fat, crude Abs was the absorbance value fibre and crude protein (Nx5.3). The carbohydrate contents were calculated by different 3) ABTS+ radical cation scavenging (carbohydrate content = 100 - summation of activity moisture, ash, crude fat, crude fibre and crude The scavenging activity of kernel extract was protein). Mineral element (Ca and K) were determined according to the modified method of determined by the wet digestion method of AOAC Long and Halliwell (2001). Briefly, 100 µl of (1995) and vitamin B1, B2 and β-carotene were different concentration of kernel extracts were determined using AOAC (2005). mixed with 900 µl of ABTS+ solution (0.7 mM of + ABTS mixed with 2.45 mM of K2S2O8 dissolved by 4. Antioxidant activities deionized water and stored in dark for 16 h; the 1) Preparation of ethanolic extract + absorbance of generated ABTS solution at 734 All kernel samples of three plants (MK, FK nm in range from 0.7-0.9 and stored in dark for 6 and TK) were extracted with 150 ml of 50% min. The absorbance was measured at 734 nm. ethanolic (ethanol: water, 50 : 50), 80% ethanolic Trolox and ascorbic acid were used as standard ethanol : water, 80 : 20) and ethanol using shaker solution. IC50 was the concentration of standard or at 200 rpm for 3 h. The solvent extracts were extracts solution with 50% ABTS+ radical separated and filtered by Whatman filter paper scavenging activity. The radical scavenging activity No.1. The solid residues were re-extracted twice was calculated using following equation. and pooled. The solvent was removed by rotary

ปีที่ 15 (ฉบับพิเศษ 1) พฤษภาคม 2561 วารสารเกษตรพระวรุณ 43 Volume 15 (SUPPL. 1) May 2018 Prawarun Agr. J. Volume 15 (SUPPL. 1) 2018, Pages 40-47

deviation (SD) (n=3). All data were analyzed by % one way analysis of variance (ANOVA) and

considered statistically significant (p < 0.05). Abs was the absorbance value

4) FRAP method Results and Discussion Ferric reducing antioxidant power (FRAP) of kernel extract was determined according to the method of Benzie and Strain (1996). Briefly, 100 µl 1. Nutritional compositions of kernel extracts were mixed with 900 µl of FRAP The nutritional composition of various reagent and incubated in water bath at 37°C for 15 kernel powder including makamong (MK), flower min. The absorbance was measured at 595 nm. fence (FK) and tamarind (TK) were shown in Table The results were expressed in terms of trolox 1. The makamong kernel powder was shown the equivalent antioxidant capacity (TEAC) in mg trolox highest quantity of energy, crude fat and ash per dry weight. (439.42±0.72 kcal, 10.64±0.07 and 2.49±0.01%) 5) Total polyphenol content than FK and TK (energy; 414.15± 0.39 and 422.8± The total polyphenol content (TPC) of 0.47, crude fat; 7.2±0.07 and 6.78±0.04, ash; kernel extract was determined according to the 2.38±0.01 and 1.77±0.01, respectively). The method of Singleton et al. (1999). Briefly, 100 µl of crudeprotein and moisture content of FK were kernel extract was mixed with 500 µl of 0.2 N revealed that significantly highest (25.29±0.21 and 3.1 ± 0.01) than MK and TK. Folic-Ciocalteu and 400 µl of 7.5% w/v Na2CO3. The mixture was incubated at room temperature for 15 min. The absorbance was measured at 765 Table 1 Nutritional composition of kernel powder nm. The results were presented in terms of gallic of makamong (MK), flower fence (FK) acid equivalent antioxidant activity (GAE) in mg and tamarind (TK) gallic acid per dry weight. 6) Total flavonoid content Composition (%) MK FK TK Energy (Kcal) 439.42 ±0.72c 414.15± 0.39a 422.8± 0.47b The total flavonoid content (TFC) of kernel a b a Moisture (g) 0.96 ±0.08 3.1 ± 0.01 1.01 ± 0.07 extracts was determined according to the method Ash (g) 2.49±0.01c 2.38±0.01b 1.77±0.01a of Chang et al. (2002). Briefly, 100 µl of kernel Crude protein 12.27±0.01a 25.29±0.21c 14.4±0.01b extract was mixed with 500 µl of 5% w/v NaNO2 (NX5.3) Crude fat (g) 10.64±0.07c 7.2±0.07b 6.78±0.04a and 400 µl of 10% w/v AlCl3 The mixture was b a c incubated at room temperature for 15 min. The Carbohydrate (g) 73.65±0.01 62.04±0.13 76.05±0.04 absorbance was measured at 415 nm. The results Values were mean ± SD followed by the capital letter, within a row (p < 0.05). were presented in terms of quercetin equivalent

(QE) in mg quercetin per dry weight. 2. Vitamin and mineral contents

The vitamin and mineral contents of various 5. Statistical analysis kernel powder including makamong (MK), flower The assay was performed in triplicate. The fence (FK) and tamarind (TK) were shown in Table results were expressed as mean± standard

44 วารสารเกษตรพระวรุณ ปีที่ 15 (ฉบับพิเศษ 1) พฤษภาคม 2561 Volume 15 (SUPPL. 1) May 2018 Prawarun Agr. J. Volume 15 (SUPPL. 1) 2018, Pages 40-47

2. The MK kernel powder was exhibited the TE g-1 DW). The 50% ethanol of all kernel extract significantly highest of quantities of -carotene was shown highest total flavonoid content and the (92.59±1.17 µgg-1) than FK and TK. The vitamin B1, strongest antioxidant activities than difference calcium and magnesium content of FK were solvent with may be correlation between polarity shown significantly highest (0.68±0.01 mg g-, of phenolic compounds in kernel extracts. Korekar 82.22±0.11mg g-1and 1.48± 0.07mg g-1) than MK et al. (2011) who revealed that total polyphenol and TK, respectively. content and free radical scavenging activity (DPPH) of apricot kernel which showed a good free radical Table 2 Vitamins and minerals content of kernel scavenging activity and enriched total polyphenol -1 powder of makamong (MK), flower contents (TPC; 0.92 -1.62 mg GAE g -1 fence (FK) and tamarind (TK) DW,DPPH;43.8-123.4 mg ml and FRAP; 154.1-243.6 -1 FeSO4.7H2O µg ml ). The ethanolic extract of Vitamin/Mineral MK FK TK mango kernel was shown higher TPC (89.9±0.23 c b a -1 -carotene 92.59±1.17 54.7±0.71 37.14±0.59 mg TAE g ) than aqueous extract (68.1±0.13 mg (µg g-1) TAE g-1) (Maisuthisakul and Gordon, 2009). b c a Vitamin B1 0.05±0.00 0.68±0.02 0.01±0.00 (mg g-1) Conclusion Vitamin B2 0.11±0.00 0.04±0.00 0.05±0.00 (mg g-1)ns Ca (mg g-1) 38.96±1.45b 82.22±0.11c 14.81±0.13a In conclusion, The MK was shown significant -1 a b a K (mg g ) 0.77±0.02 1.48± 0.07 0.82±0.02 energy, ash, vitamin B1 and free radical scavenging Values weremean ± SD followed by the sameletter, within activity. Additionally, FK exhibited significant crude a row (p < 0.05). protein and had highest the total polyphenol

contents. All kernels have shown a good source of 3. Antioxidant activities protein, fat, vitamin, mineral and nature The antioxidant activities of different antioxidant. solvent of ethanolic kernel extract (MK, FK and

TM), 50% ethanol (ethanol : water; 50:50), 80% ethanol (ethanol : water; 80 : 20) and ethanol Acknowledgements were extracted the all kernel powder and these results were shown in Table 3. The strongest free The financial was supported from Research radical scavenging activity (DPPH and ABTS+) of and development Institute, Maha Sarakham -1 Rajabhat University and Department of Chemistry, MK50 were IC50 8.80±0.03 mg ml and 3.12±0.02 mg ml-1. The FK80 was shown the significantly Faculty of Science and Technology, Maha highest total polyphenol (69.60±0.45 mg GAE g-1 Sarakham Rajabhat University for partially DW). The MK80 was shown the significantly highest supporting this research. ferric reducing antioxidant activity (4.80±0.01 mg

ปีที่ 15 (ฉบับพิเศษ 1) พฤษภาคม 2561 วารสารเกษตรพระวรุณ 45 Volume 15 (SUPPL. 1) May 2018 Prawarun Agr. J. Volume 15 (SUPPL. 1) 2018, Pages 40-47

Table 3 Total polyphenol, total flavonoid contents and antioxidant activities of kernel powder of makamong (MK), flower fence (FK) and tamarind (TK)

+ TPC TFC IC50 DPPH IC50 ABTS FRAP (mg GAE g-1 DW) (mg QE g-1 DW) (mg ml-1) (mg ml-1) (mg TE g-1 DW) MK50 26.63±0.32 37.95±0.38 8.80±0.03 3.12±0.02 4.54±0.03 MK80 20.49±0.31 22.14±0.51 14.29±0.03 3.63±0.02 4.80±0.01 MK100 29.11±0.82 2.36±0.31 60.37±0.51 13.53±0.12 1.63±0.01 FK50 25.51±0.23 9.27±0.58 26.41±0.24 2.22±0.03 2.74±0.01 FK80 69.60±0.45 1.85±0.14 34.15±0.14 6.85±0.02 2.16±0.00 FK100 20.89±0.07 0.67±0.14 80.50±0.76 23.22±0.12 1.58±0.01 TK50 65.06±0.30 12.33±0.30 18.32±0.14 6.80±0.02 2.24±0.02 TK80 64.20±0.15 6.75±0.45 30.08±0.24 7.70±0.01 1.86±0.02 TK100 20.81±0.07 0.51±0.25 90.00±0.52 24.69±0.12 1.42±0.02 MK50 = 50% ethanolic kernel extract of makamong; MK80 = 80% ethanolic kernel extract of makamong; MK100 = ethanolic kernel extract of makamong; FK50 = 50% ethanolic kernel extract of flower fence; FK80 = 80% ethanolic kernel extract of flower fence; FK100 = ethanolic kernel extract of flower fence; TK = 50% ethanolic kernel extract of tamarind; MK80 = 80% ethanolic kernel extract of tamarind; TK100 = ethanolic kernel extract of tamarind.

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by product. Food Chemi. 117(2): 332-341. Mishra, A. and Malhotra, A.V. 2009. Tamarind xyloglucan: a polysaccharide with versatile application potential. J. Mater. Chem. 19(45): 8528-8536. Sastri, B.N. 1998. The wealth of India, raw materials. New Delhi: Publication and information directorate, CSIR, 3: 13-14. Singleton, V.L., Orthofer, R. And Lamuela-Raventós, R.M. 1999. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods Enzymol. 299: 152-178. Takawale, H., Mute, V., Awari, D., Hukkeri, V.I., Mehta, P. and Vawhal, P. 2011. Screening of antiulcer activity of Caesalpinia pulcherrima L. Bark. against aspirin induced ulcer in rats. World J. Med. Sci. 6(4): 168-172. Talumatsawat, M. 2006. HuaySai trees Phetchaburi Province. Forest Herbarium. Xu, J., Yang, F., Chen, L., Hu, Y. and Hu, Q. 2003. Effect of selenium on increasing the antioxidant activity of tea leaves harvested during the early spring tea producing season. J. Agric. Food Chem. 51(4): 1081- 1084.

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