PROPOSAL BANTUAN PENULISAN KARYA ILMIAH UNTUK DITERBITKAN PADA JURNAL INTERNASIONAL T AHUN ANGG AR AN 2016
JUDUL MANUSKRIP In Vitro Antidiabetic Activity of Two Seaweed from Yogyakarta Beach of Indonesia
Tim Penulis Amir Husni, Tiara Pratiwi, Ustadi, Agung Giri Samudra, dan Agung Endro Nugroho
Fakultas Pertanian UNIVERSITAS GADJAH MADA 2016
PROPOSAL
A. Latar Belakang
Dalam rangka deseminasi hasil penelitian maka data penelitian perlu ditulis dalam bentuk naskah publikasi ilmiah. Salah satu penelitian yang telah selesai dilaksanakan adalah In Vitro Antidiabetic Activity of Two Seaweed from Yogyakarta Beach of Indonesia. Oleh karena itu dalam kesempatan ini penulis akan mempublikasikan hasil penelitian tersebut dalam jurnal ilmiah bereputasi Internasional terindeks Scopus yaitu Pertanika Journal of Tropical Agricultural Science.
B. Tujuan Penulisan
Tujuan penulisan jurnal adalah sebagai sarana mendeseminasikan hasil penelitian dengan judul In Vitro Antidiabetic Activity of Two Seaweed from Yogyakarta Beach of Indonesia.
C. Kontribusi Penelitian
Penulisan naskah publikasi ini didasarkan pada hasil penelitian dari program Penelitian Unggulan Universitas Gadjah Mada dengan dana yang diperoleh dari DIPA Desentralisasi Universitas Gadjah Mada tahun 2013.
D. Manfaat yang diharapkan
Dengan terbitnya publikasi hasil penelitian ini maka akan diperoleh manfaat baik bagi individu penulis maupun Universitas Gadjah Mada serta masyarakat secara luas.
E.Target Pembaca dan Penerbit
Target pembaca untuk naskah publikasi ini adalah para peneliti/pemerhati bidang pertanian dan perikanan khususnya tentang pemanfaatan rumput laut secara umum dan aktivitas bioaktifnya maupun para peneliti/pemerhati masalah bahanaktif anti-diabetes, Naskah publikasi ini diharapkan dapat terbit pada Pertanika Journal of Tropical Agricultural Science..Jurnal ini merupakan jurnal Internasional yang diterbitkan oleh UPM Press dan merupakan jurnal yang open access yang terindeks Scopus.
Lampiran BIODATA PENGUSUL BANTUAN PENULISAN KARYA ILMIAH UNTUK DITERBITKAN PADA JURNAL INTERNASIONAL T AHUN ANGG AR AN 2016 I. IDENTITAS DIRI 1.1 Nama Lengkap (dengan gelar) 1.2 Jabatan Fungsional 1.3 1.4 1.5
NIP/NIK/No. identitas lainnya Tempat dan Tanggal Lahir Alamat Rumah
1.6 1.7 1.8
Nomor Telepon/Faks Nomor HP Alamat Kantor
1.9 1.10 1.11
Nomor Telepon/Faks Alamat e-mail Lulusan yg telah dihasilkan
1.12 Mata Kuliah yg diampu
II. RIWAYAT PENDIDIKAN S-1 2.1 Program: 2.2 Nama PT Universitas Gadjah Mada 2.3 Bidang Ilmu Ilmu Perikanan
2.4 TahunMasuk 2.5. Tahun Lulus
Dr.Sc. Amir Husni, S.Pi., M.P. Lektor Kepala
L
19700921 199803 1 002 Karanganyar, 21 September 1970 Temanggal II 03/01 Purwomartani Kalasan Sleman Yogyakarta 55571 0274-551218 085743255664 Jl. Flora Gedung A4 Bulaksumur Yogyakarta 55281 0274-551218
[email protected] S-1= 35 orang ; S-2= 2 orang; S-3= orang 1. Proses Thermal Hasil Perikanan 2. Refrigerasi Hasil Perikanan 3. Manajemen Limbah Industri Perikanan 4. Mikrobiologi 5. Gizi Ikani 6. Toksikologi dan Higiene 7. Pengendalian Mutu Hasil Perikanan
S-2 Universitas Gadjah Mada Ilmu & Teknologi Pangan
1990 1996
1996 1999
S-3 Gangneung-Wonju National University Applied Marine Biotechnology & Engineering 2006 2009
III PENGALAMAN PENELITIAN (bukan skripsi/tesis/disertasi) No
Tahun
Judul Penelitian
1
2010
2
2011
3
2012
Pengembangan Metode Ekstraksi Alginat dan Formulasinya Sebagai Bahan Pengental pada Tekstile Printing Peningkatan Daya Simpan Filet Nila Merah Menggunakan Ekstrak Rumput laut dari Gunungkidul Pengembangan Pangan Fungsional Berbasis Komoditas Lokal yang Diperkaya dengan Rumput Laut untuk Mendukung
Pendanaan Sumber Jumlah (Rp) DIKTI 73.000.000
Fakultas Pertanian UGM Dikti
10.000.000 96.000.000
4
2012
5
2013
6
2013
7
2013
8
2013
9
2014
10
2014
11
2015
IV
Ketahanan Pangan Mikroemulsi Ekstrak Sargassum sp. Sebagai Penghambat Kerusakan Minyak Ikan Pemanfaatan Rumput Laut dari Pantai Gunungkidul untuk Peningkatan Daya Simpan dan Keamanan Ikan Kembung Pengembangan Pangan Fungsional Berbasis Komoditas Lokal yang Diperkaya dengan Rumput Laut untuk Mendukung Ketahanan Pangan (Lanjutan I) Aktivitas Antistres dan Antioksidan RumputLaut Coklat (Phaeophyceae) Mikroemulsi Ekstrak Sargassum sp. Sebagai Penghambat Kerusakan Minyak Ikan Aktivitas Antistres dan Antioksidan RumputLaut Coklat (Phaeophyceae) (Lanjutan I) Pengembangan Pangan Fungsional Berbasis Komoditas Lokal yang Diperkaya dengan Rumput Laut untuk Mendukung Ketahanan Pangan (Lanjutan II) Aktivitas Antistres dan Antioksidan RumputLaut Coklat (Phaeophyceae) (Lanjutan II)
Dikti
Fakultas Pertanian UGM Dikti
95.000.000
8.000.000 95.000.000
Dikti
100.000.000
Dikti
94.000.000
Dikti
96.000.000
Dikti
92.000.000
Dikti
96.000.000
PENGALAMAN PENULISAN ARTIKEL ILMIAH DALAM JURNAL Volume/ No. Tahun Judul Artikel Ilmiah Nomor 1 2012 Pengembangan Metode Ekstraksi 32(1) Alginat dari Rumput Laut Sargassum sp. Sebagai Bahan Pengental 4(5) 2 2012 Peningkatan daya simpan filet nila merah menggunakan ekstrak rumput laut dari Gunungkidul 3 2013 Ekstrak Sargassum sp. Sebagai 8(2) antioksidan dalam Sistem Emulsi Minyak Ikan Selama Penyimpanan pada Suhu Kamar 4
2013
The Use of Gracilaria sp. Extract on Refrigerated Tilapia Fillet
13(7)
5
2014
34(3)
6
2014
7
2014
Penggunaan ekstrak rumput laut Padina sp. untuk peningkatan daya simpan filet nila merah. Inhibitory Activity of α-Amylase and αGlucosidase by Padina pavonica Extracts Aktivitas Antioksidan Padina sp. pada Berbagai Suhu dan Lama Pengeringan
14(8) 9(2)
Nama Jurnal Agritech Jurnal Penelitian dan Pengembangan Jurnal Pascapanen dan Bioteknologi Kelautan dan Perikanan Journal of Biological Sciences Agritech Journal of Biological Sciences Jurnal Pascapanen dan Bioteknologi
8
2015
9
2015
10
2015
Peningkatan Daya Simpan Ikan Kembung dengan Ekstrak Etanolik Padina sp. selama Penyimpanan Suhu Kamar Potensi Ekstrak Rumput Laut Eucheuma denticulatum Sebagai Agen Antidiabetes Aktivitas Antioksidan dan Tingkat Penerimaan Konsumen pada Yoghurt yang Diperkaya dengan Ekstrak Sargassum polycystum
18(1)
7(1)
18(2)
Kelautan dan Perikanan Jurnal Pengolahan Hasil Perikanan Indonesia Jurnal Penelitian dan Pengembangan Jurnal Pengolahan Hasil Perikanan Indonesia
V. PENGALAMAN MERUMUSKAN KEBIJAKAN PUBLIK/REKAYASA SOSIAL LAINNYA No.
Tahun
1
2011
Judul/Tema/Jenis Rekayasa Sosial Lainnya yang Telah Diterapkan
Tempat Penerapan
Respons Masyarakat
Desain Pengolahan Ikan Secara Ergonomis
Provinsi D.I. Yogyakarta
Sangat baik
VI. PENGALAMAN MENULIS BUKU 2014, Pengendalian Mutu Hasil Perikanan, UGM Press. Semua data yang saya isikan dan tercantum dalam biodata ini adalah benar dan dapat dipertanggungjawabkan secara hukum. Apabila di kemudian hari ternyata ditemukan ketidaksesuaian dengan sebenarnya, saya sanggup menerima resikonya. Demikian biodata ini saya buat dengan sebenarnya untuk memenuhi salah satu persyaratan dalam pengajuan Insentif Jurnal Karya Ilmiah Internasional.
Yogyakarta, Pengusul,
Desember 2015
(Dr. Amir Husni, S.Pi., M.P.)
In Vitro Antidiabetic Activity of Two Seaweed from Yogyakarta Beach of Indonesia
Husni, A.1*, Pratiwi, T.1,2, Ustadi1, Samudra, A. G.3,4, Nugroho, A. E.3 1
Department of Fisheries Faculty of Agriculture Universitas Gadjah Mada, Jalan Flora GedungA4 Bulaksumur Yogyakarta 55281, Indonesia 2 Ministry of Marine and Fisheries, 15 Floor Mina Bahari Building 3, Jalan. Medan Merdeka Timur 16 Jakarta, Indonesia 3 Department of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy Universitas Gadjah Mada, Sekip Utara Yogyakarta 55281 Indonesia 4 Akademi Farmasi Al-Fatah, Jalan Indragiri Gang 3 Serangkai Padang Harapan Bengkulu, Indonesia
ABSTRACT The objective of this research was to determine the potential of polyphenols and phlorotannins extract from Sargassum hystrix and Eucheuma denticulatum in inhibiting α-amylase and α-glucosidase. Polyphenols were extracted using 50% methanol, and phlorotannins was extracted using methanol, and the non-lipid layer was separated by using distilled water, methanol, and chloroform and then partitioned using ethyl acetate twice. The total content of polyphenols and phlorotannins were analyzed. Both types of the compounds were tested to know their ability to inhibit α-amylase and α-glucosidase activity. Total phenols content of S. hystrix and E. denticulatum were obtained 3.17 g GAE/100 g extract and 0.33 g GAE/100 g extract, respectively. Total phlorotannin content of S. hystrix and E. denticulatum were obtained 0.02 g PGE/100 g extract and 0.02 g PGE/100 g extract, respectively. The results showed that polyphenols S. hystrix (IC50 = 0.58±0.01mg/mL) can inhibit α-amylase similar to acarbose (IC50 = 0.53±0.00 mg/mL) and phloroglucinol (IC50 = 0.56±0.01 mg/mL). Inhibitory activity of Polyphenols S. hystrix (IC50 = 0.59±0.02 mg/mL) in inhibiting α-glucosidase also similar to acarbose (IC50 = 0.61±0.01 mg/mL) and phloroglucinol (IC50 = 0.56±0.05 mg/mL). Keywords: α-amylase, α-glucosidase, antidiabetic activity, Eucheuma denticulatum, Sargassum hystrix
INTRODUCTION Diabetes mellitus (DM) is a group of metabolic disorder characterized by hyperglycemia resulting from defects in insulin secretion and/or insulin action (ADA, 2012). Chronic hyperglycemia in diabetes related to long-term damage, dysfunction of several organs of
the body (Elekofehinti et al., 2013). WHO (2012) has been reported that approximately 346 million people in the world suffer DM. The International Diabetes Federation (IDF, 2012) approximated that in 2030, people with diabetes would rise to 438 million. Marine alga is a potential source of bioactive secondary metabolites with high developed as a new pharmaceutical agent. Marine algae have content such as polysaccharides, proteins, peptides, amino acids, lipids, minerals, and some vitamins. Bioactive compounds from brown marine algae are potential as an antitumor, antifungal, antiviral, antioxidant, antihypertensive and antidiabetic (Gamal, 2010; Husni et al., 2014). The polyphenol content of marine algae can be used as cosmetic and has pharmacological effects as antioxidants, antibiotics, anti-inflammatory, hypo-allergenic, antibacterial, and antidiabetic (Holdt & Kraan, 2011). Ye et al. (2009) reported that ethanol extract of brown marine algae Sargassum pallidum has antioxidant activity. Nwosu et al. (2011) stated that phenol extract of seaweed Palmaria and Ascophyllum able to inhibit the activity of αamylase and α-glucosidase that could lower blood glucose levels. Marine algae also have a high content of antioxidants and can be used to ward off free radicals that increase due to the condition of hyperglycemia in diabetic people (Firdaus et al., 2010). Until now, research information on the antidiabetic activity of two marine algae Sargassum hystrix and Eucheuma denticulatum have not been found. The objective of this study was to determine the activity of S. hystrix and E. denticulatum extracts in inhibiting the activity of α-amylase and α-glucosidase as an antidiabetes agent.
MATERIALS AND METHODS Materials The main material was used in this study was seaweed S. hystrix & E. denticulatum obtained from coastal of Gunungkidul Yogyakarta Indonesia. The materials were used for
the extraction of polyphenols and phlorotannin namely: methanol (E. Merck), sodium carbonate (E. Merck), Folin-Ciocalteu's reagent (E. Merck), and acid gallate (E. Merck). The materials were used to analyze the α-amylase inhibition activity: α-amylase from Bacillus sp. type II-A (Sigma-Aldrich), 3,5-dinitrosalisilat (DNS) acid (Sigma-Aldrich), and essential oils (E. Merck). The materials to analyze the α-glucosidase inhibition activity was: α-glucosidase from Saccharomyces cerevisiae type I (Sigma-Aldrich), p-nitrofenil-αd-glucophiranoside (Sigma-Aldrich), and acarbose (Bayer Pharmaceuticals).
Marine Algae Extractions In this study, the extraction of seaweed has been made to obtain an extract containing polyphenols and phlorotannin. The extraction of the seaweed S. hystrix and E. denticulatum to get polyphenols using a modification of the method Zhang et al. (2007). Dry powder weighed 40 g and was put in a closed Erlenmeyer aluminum foil. Then added 300 mL of 50% methanol adjust pH 2 and stirred for 4 h and then allowed to stand for 24 h. The solution was filtered and then centrifuged at a speed of 3500 rpm for 20 min and then separate the supernatant. Supernatant evaporated at a speed of 30 rpm until concentrated and then freeze drying. Results freeze dry were weighed for yield calculation and stored at -20 °C. Phlorotannin extract obtained by partitioning using a modified method of Chowdhury et al. (2011). Polyphenol powder 5 g were weighed and put in a closed Erlenmeyer aluminum foil. Afterward added 40 mL of methanol and stirred for two h and then allowed to stand for 24 h and then added 20 mL of chloroform while stirring for 20 min. The solution was centrifuged at a speed of 3500 rpm for 20 min then separated supernatant. Supernatant plus aqua bidest many as 15 mL while stirring for 10 min and then will form lipid and non-lipid layer. Non-lipid layer that sits at the top is taken and added 25 mL of
ethyl acetate and stirred for 30 min. Subsequently, the mixture was evaporated, freezedried and stored at -20 °C until use in further research.
Measurement of Total Polyphenol Content The content of total phenols in seaweed was analyzed using a modified method of Zhang et al. (2007). Gallic acid as standard made into several concentrations ranging from 0 to 400 mg mL-1. Polyphenol extracts made serial dilution to a concentration of 3.125 to 200 mg mL-1. Then 500 mL of each concentration of 200 mL was taken and put into a test tube. Further plus 1000 mL Folin-Ciocalteu reagent and incubation for 5 min. Then added 800 mL of 20% Na2CO3 and incubated in the dark room and the room temperature (27 °C) for 75 min. Afterward, he supernatant was taken and immediately read the absorbance at 750 nm wavelength.
Measurement of Total Phlorotannin Content The content of total phlorotannin of seaweed was analyzed using a modified method Koivikko et al. (2005). Phlorotannin extract 0.1 g, macerated with 200 mL of 85% ethanol (1: 2) in the dark for eight h, and then made into a dilution series of concentration, 125 to 200 mg mL-1. Then a mixture of 500 mL pipettes and put into a test tube and then added 500 mL of Folin-Ciocalteu reagent and one mL 20% Na2CO3, and then left for 3 min. Furthermore, the solution in a test tube incubated in the dark room temperature of 27 °C for 45 min, then centrifuged for 10 min at a speed of 3,500 rpm. The supernatant was taken and immediately read the absorbance at 730 nm wavelength.
Inhibition of α-Amylase Activity
Inhibitory activity of α-amylase was determined by measuring changes in 3.5dinitrosalicylate acid into nitro-aminosalicylate acid (catalyzed by α-amylase) using spectrophotometry (Apostolidis et al., 2011). Test solution consisting of 25 mL sample extract at various concentrations and 25 mL of sodium phosphate buffer 0.02 M (pH 6.9 with 0.006 M NaCl) containing 13 U mL-1 of α-amylase. The test solution was mixed and incubated at 37 °C for 10 min. After pre-incubation, added 25 mL of soluble starch 1% in sodium phosphate buffer 0.02 M and incubated at 37 °C for 10 min. Then proceed with the addition of 50 mL 96 mM 3,5-dinitrosalisilat acid (DNS) as the color reagent solution and incubated for 5 min in boiling water. The solution was cooled to room temperature and the absorbance of the solution was read at a wavelength of 550 nm. Absorbance values were obtained then used to calculate the percentage inhibition of the enzyme.
where: K= Absorbance of control-blank S1= Absorbance of sample with enzyme S0= Absorbance of sample without enzyme Inhibition of α-Glucosidase Activity Inhibitory activity test of α-glucosidase was taken out according to the method of Mayur et al. (2010). Test solution consisting of 50 mL 0.1 M phosphate buffer (KH2PO4) pH 7, 25 mL substrate 0.5 mM p-nitrophenyl-α-D-glucopyranoside (PNP-G), 10 mL sample extract at various concentrations, and 25 mL α-glucosidase at a concentration of 0.2 U mL-1. The solution was mixed and incubated at 37 °C for 30 min. The reaction was stopped by addition of 100 mL 0.2 M Na2CO3. Inhibition of enzyme activity was measured by the amount of p-nitrophenyl formed by measuring absorbance using a microplate reader at a wavelength of 405 nm. Absorbance values were obtained then used to calculate the percentage inhibition of the enzyme.
where: K= Absorbance of control-blank S1= Absorbance of sample with enzyme S0= Absorbance of sample without enzyme Statistical Analysis The data in this study were extract concentration versus percent inhibition of the enzyme. The data were plotted to obtain the regression equation. The IC50 activity value of S. hystrix and E. denticulatum extracts against α-amylase and α-glucosidase were achieved from the regression equation. The IC50 values were analyzed statistically with one-way analysis of variance using the SPSS (Statistical Package for Social Sciences) at 95%.
RESULTS AND DISCUSSION Total Polyphenol and Phlorotannin Contents The data of polyphenols and phlorotannins extracts along with the content of each compound were shown in Table 1. The total phenols content of S. hystrix and E. denticulatum were obtained 3.17 g GAE 100g-1 extract and 0.33 g GAE 100g-1 extract, respectively.
Damongilala et al. (2013) reported that extraction using 60% methanol
resulted in total phenol 5.87±0:15 mg GAE g-1 extract while methanol 70% and 80% resulted in total phenol content of 4.98±0:02 mg GAE g-1 extract and 5.76±0.11 mg GAE g1
extracts, respectively. The results of the analysis of total phenol are consistent with
research Kumar et al. (2011). The total phenols were biggest with the brown algae (43.75 to 55.23 mg PGE g-1 extract) than red algae (22.79 to 65.60 mg PGE g-1 extract) and green algae (32,57 to 56.22 mg PGE g-1 extract). Research Cox et al. (2010) showed that brown algae (Himanthalia elongata) have a higher total phenol was 151.33±6.75 mg GAE g-1 extract. Zubia et al. (2007) reported that the total phenols content of the Padina
gymnospora was 5.58±0.30%. Damongilala et al. (2013) showed that P. pavonica was extracted using 60% methanol resulted in the total phenols content of 4.98 to 5.87 mg GAE g-1 extract. The differences in total phenols content could be influenced by intrinsic factors including age, type, etc. and extrinsic factors namely tidal cycles, salinity, etc. (Connan et al., 2007). Table 1 showed that total phlorotannin content of S. hystrix and E. denticulatum were obtained 0.02 g PGE 100g-1
extract and 0.02 g PGE 100g-1 extract, respectively.
According to Koivikko et al. (2008), only the brown algae that contain phlorotannin. However, based on this study, detected phlorotannin content of the red alga E. denticulatum although phlorotannin content no higher than S. hystrix. The results showed that phlorotannin content was lower than phlorotannin content in S. echinocarpum (6.75 mg PGE g-1 extract) extracted using methanol (Firdaus, 2011). The phlorotannin content of the brown algae Eisenia bicyclis and E. kurome were 3.1, and 3.0%, respectively (Shibata et al., 2004). Phlorotannin content it is specific to each species. According to Jormalainen & Honkanen (2004), the phlorotannin concentration of the algae influenced by several factors, namely age, species, and environment (location, season, waves, presence or absence of light, salinity, UV radiation, the presence or absence of herbivores, and nutrients).
In Vitro Antidiabetic Activity of Two Seaweeds
TABLE 1 Yield and total content of polyphenols and phlorotannin extracts from Sargassum hystrix and Eucheuma denticulatum Extracts
Sample Sargassum hystrix
Yield Total content
Eucheuma denticulatum
Yield Total content
Polyphenols 15.47% 3.17 g GAE 100g-1
Phlorotannin 2.36% 0.021 g PGE 100g-1
12.40%
8.24%
0.33 g GAE 100g-1
0.018 g PGE 100g-1
Fig. 1: Effect sample concentration (▲: polyphenol Sargassum hystrix, ×: polyphenol Eucheuma denticulatum, ○: Phlorotannin Sargassum hystrix, ●: Phlorotannin Eucheuma denticulatum) and control (◊: acarbose, ■: phloroglucinol) on inhibition activity of αamylase. Inhibition of α-Amilase Activity Based on Fig. 1 showed that the highest inhibitory activity was obtained in polyphenols of S. hystrix (94.09%). Meanwhile, at the same concentration, acarbose, polyphenols of E. denticulatum, and phloroglucinol showed a smaller inhibitory activity (88.81%, 85.84%,
In Vitro Antidiabetic Activity of Two Seaweeds
and 85.92%, respectively) and the activity was not significantly different compare to phlorotannin of S. hystrix (68.39%) and phlorotannin of E. denticulatum (69.02%). Polyphenols of S. hystrix at these concentrations showed higher inhibitory activity compare to polyphenols of E. denticulate, phlorotannin of S. hystrix and phlorotannin of E. denticulatum. According to Kunyanga et al. (2012), phenolic compounds able to bind to the active site of alpha-amylase. Bioactive components group of phenolics such as anthocyanins, flavonols, flavones, flavanones, gallic acid, vanillic acid, quercetin and trans-cinnamic has been reported to have inhibitory activity against the activity of αamylase (Chethan et al., 2008; Hanhineva et al., 2010). Their ability of both seaweed, in inhibiting alpha-amylase was supported by research Firdaus et al. (2010) in the use of Sargassum echinocarpum as antihyperglycemic agents and Lamella et al. (1989) has also been reported that Eucheuma sp. having hypoglycemic activity. Lopes et al. (2012) reported that polyphenols consist of flavonoid or gallic acid polymer in terrestrial plants while the seaweed consisting of units of phloroglucinol (1,3,5-trihydroxybenzene). Table 2 showed the IC50 of inhibition activity of α-amylase by S. hystrix and E. denticulatum extracts, acarbose, and phloroglucinol. The IC50 values from the sixth test samples ranging from the smallest to largest were acarbose, phloroglucinol, polyphenols of S. hystrix, phlorotannin of E. denticulatum, polyphenols of E. denticulatum, and phlorotannin of S. hystrix. There are not significant between IC50 values of acarbose, phloroglucinol, and polyphenol of S. hystrix. The IC50 values showed higher activity compare to another marine algae but also lower than Ascophyllum nodosum. Senthilkumar & Sudha (2012), informed that IC50 value of the water extracts from marine algae S. policystum, R. corticata, and G. lactuca in inhibiting of α-amylase was 60, 67, and 82 µg mL-1, respectively. Nwosu et al. (2011) reported that methanol extract of A.nodosum has a smaller IC50value (0.1 µg mL-1).
In Vitro Antidiabetic Activity of Two Seaweeds
Inhibition of α-Glucosidase Activity The ability of each sample to inhibit the activity of α-glucosidase were presented in Fig. 2. The inhibition activity α-glucosidase showed that acarbose and polyphenols of S. hystrix have similar ability to inhibit α-glucosidase activity, followed by phloroglucinol, polyphenols of E. denticulatum, phlorotannin of E. denticulatum, and phlorotannin of S, hystrix. IC50 value of the six samples that were tested were able to inhibit the activity of αglucosidase from largest to smallest were phloroglucinol, polyphenols S. hystrix, acarbose, phlorotannin S. hystrix, phlorotannin E. denticulatum, and polyphenols E. denticulatum (Table 2). The ability of polyphenols to inhibit α-glucosidase in the digestive tract and activation of glucose uptake will be able to lower blood glucose (Zhang et al., 2007; Apostolidis & Lee, 2011). Anthosianin especially polyphenols, flavonols, proanthosianin, and phenolic acids significantly suppress the increase in blood glucose and reduce the rate of digestion of sucrose and glucose absorption in the intestine (Wilson et al., 2008). The workings of a polyphenol similar to acarbose, which extends the time revamp of carbohydrates and inhibit the absorption of glucose (You & Barnett, 2004).
In Vitro Antidiabetic Activity of Two Seaweeds
Fig. 2: Effect sample concentration (▲: polyphenol Sargassum hystrix, ×: polyphenol Eucheuma denticulatum, ○: Phlorotannin Sargassum hystrix, ●: Phlorotannin Eucheuma denticulatum) and control (◊: acarbose, ■: phloroglucinol) on inhibition activity of αglucosidase
TABLE 2 Inhibitory activity (IC50) of polyphenol and phlorotannin extracts from Sargassum hystrix, Eucheuma denticulatum, acarbose, and phloroglucinol against α-amylase and α-glucosidase Inhibitors Acarbose Phloroglucinol Polyphenols Sargassum hystrix Polyphenols Eucheuma denticulatum Phlorotannin Sargassum hystrix Phlorotannin Eucheuma denticulatum
IC50of α-amylase (mg mL-1) 0.53±0.00a 0.56±0.01a 0.58±0.01a 1.43±0.19
b
3.29±0.12d 1.92±0.14
c
IC50of α-glucosidase (mg mL-1) 0.61±0.01a 0.56±0.05a 0.59±0.02a 1.43±0.19d 0.78±0.04b 0.86±0.06c
Phenols are one of the bioactive components that can inhibit the action of α-amylase and α-glucosidase. Polyphenols can inhibit the enzyme in the breakdown of carbohydrates into glucose. The content of phenol has an inhibitory effect on α-amylase through bond hydroxylation and ring substitution on β. The principle is similar to acarbose inhibition, i.e.,
In Vitro Antidiabetic Activity of Two Seaweeds
by generating delays and disaccharide carbohydrate hydrolysis and absorption of glucose and inhibits the metabolism of sucrose into glucose and fructose (You et al., 2012). Beside that phlorotannin is one of the phenolic components which can also inhibit the work of the αamylase and α-glucosidase. As polyphenols, phlorotannin inhibits enzymes work in the breakdown of carbohydrates into glucose. The principle also is similar to acarbose inhibition; that is to produce a delay hydrolysis and absorption of carbohydrates and disaccharides inhibit the metabolism of glucose and sucrose into glucose and fructose (You et al., 2012).
CONCLUSION Seaweed
S. hystrix have more potential as an antidiabetic substance compare to E.
denticulatum. The results showed that polyphenols S. hystrix (IC50 = 0.58±0.01 mg mL-1) can inhibit α-amylase similar to acarbose (IC50 = 0.53±0.00 mg mL-1) and phloroglucinol (IC50 = 0.56±0.01 mg mL-1). Inhibitory activity of polyphenols S. hystrix (IC50 = 0.59±0.02 mg mL-1) in inhibiting α-glucosidase also similar to acarbose (IC50 = 0.61±0.01 mg mL-1) and phloroglucinol (IC50 = 0.56±0.05 mg mL-1).
ACKNOWLEDGEMENTS This research was supported Institue for Research and Community Services via Research Grants Flagship of Universitas Gadjah Mada through DIPA Universitas Gadjah Mada 2013 Number LPPM-UGM/1406/LIT/2013.
In Vitro Antidiabetic Activity of Two Seaweeds
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