Various solvent effects on phytochemical constituent profiles, analysis of antioxidant and antidiabetic activities of Hopea parviflora

Journal Pre-proof Various solvent effects on phytochemical constituent profiles, analysis of antioxidant and antidiabetic activities of Hopea parviflora Ranjitha Venkatachalam, Kandasamy Kalimuthu, Vajjiram Chinnadurai, Mythili Saravanan, Ramalingam Radhakrishnan, Rajasree Shanmuganathan, Arivalagan Pugazhendhi

PII:

S1359-5113(19)31126-2

DOI:

https://doi.org/10.1016/j.procbio.2019.10.025

Reference:

PRBI 11813

To appear in:

Process Biochemistry

Received Date:

30 July 2019

Revised Date:

6 October 2019

Accepted Date:

22 October 2019

Please cite this article as: Venkatachalam R, Kalimuthu K, Chinnadurai V, Saravanan M, Radhakrishnan R, Shanmuganathan R, Pugazhendhi A, Various solvent effects on phytochemical constituent profiles, analysis of antioxidant and antidiabetic activities of Hopea parviflora, Process Biochemistry (2019), doi: https://doi.org/10.1016/j.procbio.2019.10.025

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Various solvent effects on phytochemical constituent profiles, analysis of antioxidant and antidiabetic activities of Hopea parviflora. Ranjitha Venkatachalam a, Kandasamy Kalimuthu a,*, Vajjiram Chinnadurai b, Mythili Saravanan c, Ramalingam Radhakrishnan d, Rajasree Shanmuganathan e, Arivalagan Pugazhendhi f a

PG and Research Department of Botany, Government Arts College (Autonomous),

Coimbatore – 641 018, Tamil Nadu, India. Department of Botany, Sri Vidya Mandir Arts and Science College, Katteri, Uthangarai,

Krishnakiri – 636 902. Tamil Nadu, India c

PG and Research Department of Biotechnology, Hindusthan College of Arts and Science,

Nava India, Coimbatore – 641 028, Tamil Nadu, India d

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b

Department of Microbiology, Karpagam Academy of Higher Education, Coimbatore, Tamil

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Nadu, India

Institute of Research and Development, Duy Tan University, Da Nang 550000, Viet Nam

f

Innovative Green Product Synthesis and Renewable Environment Development Research

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e

Group, Faculty of Environment and Labour Safety, Ton Duc Thang University, Ho Chi Minh

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City, Viet Nam. Email id: [email protected]

*Corresponding Author Address

Assistant Professor

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Dr. Kandasamy Kalimuthu

PG and Research Department of Botany

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Government Arts College

Affiliated to Bharathiar University

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Coimbatore, Tamil Nadu India.

Email id: [email protected]

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Graphical abstract

Highlights 

In vitro antidiabetic study of Hopea parviflora.



Hopea parviflora has the highest level of polyphenol and free radical scavenging activity.



Methanol extracts showed the highest percentage reduction in blood glucose levels.

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

Hopea parviflora - potential candidate for cost effective antidiabetic with less side effects.

Abstract The current research has been designed to assess the phytochemical composition, antioxidant and antidiabetic properties of Hopea parviflora, sequentially extracted with petroleum ether, chloroform, ethyl acetate, ethanol and methanol. All the five extracts were tested for qualitative and quantitative phytochemicals. DPPH, Superoxide, FRAP, ABTS and metal chelating antioxidant activities were evaluated. Antidiabetic potentials of all the five extracts

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were tested using standard in vitro α- amylase and α - glycosidase inhibition assays. Qualitative phytochemical screening showed the presence of alkaloids in all the extracts except petroleum ether and ethyl acetate. Steroids were present in the petroleum ether, ethyl acetate and chloroform extracts whereas glycosides were present in all the extracts, except

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ethanol. The total phenol, flavonoid, tannin and saponins contents varied from solvent to solvent, with the highest values being 18.9, 18.2, 0.98 and 39.9 mg/mL, respectively.

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Methanolic extract showed the highest antioxidant activities in DPPH, FRAP and superoxide assays. Moreover, effective results were observed for the ethanol and ethyl acetate extracts in the ABTS and metal chelating assays. The methanolic extract showed potential antidiabetic

Abbreviations -

FRAP

-

ABTS

Ferric reducing antioxidant power

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2, 2-azino-bis (ethylbenzene-thiazoline-6-sulfonic acid)

-

Ethylene diamine tetra acetic acid

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EDTA

1, 1-Diphenyl-2-picrylhydrazyl

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DPPH

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inhibition assays, respectively.

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activities with the IC50 values of 230.2 and 308.2 µg/mL in α- amylase and α -glycosidase

K2S2O8

-

Potassium persulfate

IC

-

Inhibition concentration

Keywords: Hopea parviflora; Antioxidant; Antidiabetic; α-Amylase; α-Glycosidase.

1. Introduction

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In recent years, usage of medicinal plants has gained significance in primary health care needs all over the world. Medicinal plants are the backbone of the traditional medical practice as most of the modern drugs and their derivatives are produced from these plants [1]. Plant based antioxidants such as polyphenols have gained substantial attention due to their potential medicinal benefits [2, 3]. Natural plant based antioxidants either in the form of crude extract or individual phytocompounds and their derivatives are very efficient in preventing cell damaging processes caused by oxidative stress [4]. The antioxidant property of polyphenol compounds is primarily due to their redox potential, which enables them to act as singlet oxygen quenchers, metal chelators, reducing agents and hydrogen donors [5].

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The plant polyphenols aid in the absorption and neutralization of free radicals, inducing expression of peroxides, singlet and triplet oxygen quenching. The human body has a natural antioxidant mechanism, which aids

in various biological functions such as

antiaging responses, antimutagenic and anticarcinogenic properties [6, 7]. Any change or defect in the antioxidant mechanism results in oxidative stress, which leads to the

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development of several contagious diseases. The commercially available synthetic antioxidant drugs may cause several side effects [6, 8].

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DM (Diabetes mellitus) is a metabolic disorder with multiple symptoms, which can be characterized by chronic hyperglycemia or hypoglycemia together with disturbances in

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carbohydrate, fat and protein metabolisms, resulting in insulin deficiency or increased insulin levels [9, 10]. According to the WHO report, there were 171 million diabetic cases in 2000, which will significantly increase up to 366 million by 2030 [11]. Currently available

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treatment methods have various side effects, which affect the quality of the patient’s life. Hence, the plant based therapy is gaining more importance [12]. Dipterocarpaceae is the most important family of tropical forest trees with more

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economic values. This family comprises of 17 genera with 500 species [13]. Hopea is a significant genus with 100 species. Hopea parviflora is an endemic tree distributed all along

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the tropical evergreen forest of the Western Ghats [14]. The tree is well known for its medicinal importance and timber value. H. parviflora contains several bioactive chemical compounds such as alkaloids, saponins, tannins, phenolics, steroids and flavonoids [15]. Certain important plant based compounds such as stilbenoids and resveratrol were isolated from the H. parviflora bark. The aim of the present study was to evaluate the phytochemical, antioxidant and antidiabetic activities of the various solvent extracts of H. parviflora leaves.

2. Materials and methods 4

2.1 Plant collection and extract preparation H. parviflora leaves were collected from Aannaikatti, Western Ghats and were authenticated by the Botanical Survey of India, Coimbatore, Tamil Nadu. The extraction was done as successive extraction using a Soxhlet apparatus. Various polar solvents such as petroleum ether (HPLPE), chloroform (HPLC), ethyl acetate (HPLEA), ethanol (HPLE) and methanol (HPLM) were used for Soxhlet extraction.

2.2 Phytochemical analysis 2.2.1 Phytochemical qualitative analysis

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Various solvent extracts of H. parviflora leaves were tested for various qualitative phytochemical constituents. The major secondary metabolites such as alkaloids, flavonoids, tannins, steroids, triterpenoids, saponins, glycosides, gum and mucilages, fixed oils and anthraquinones were screened using standard procedures [16].

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2.2.2 Phytochemical quantitative analysis 2.2.2.1 Total phenolic content

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The various solvent extracts of H. parviflora leaves were examined for the total phenolic contents. About 20 μg of each extract was made up with 1.0 mL distilled water, the

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solution was mixed with 0.5 mL Folin-ciocalteu phenol reagent and 2.5 mL of 20% sodium carbonate solution was added. Then, the mixture was kept in dark for 40 min. Later, the absorbance was recorded at 725 nm and was compared to the absorbance of the standard

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gallic acid by Siddhuraju and Becker [17].

2.2.2.2 Total flavonoid content

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Total amount of flavonoids of each extracts were determined by using the method of Zhishen et al. [18]. 0.5 mL of each extracts were diluted with 2 mL distilled water and then

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solution was mixed with 5% NaNO3 solution and allowed for 6 min reaction time. After the reaction, 0.15 mL of 10% aluminium chloride and 4% of 2 mL sodium hydroxide solution were added then final volume was made with 5.0 mL distilled water. Finally, the mixture was incubated for 15 min at 37 ºC and absorbance was recorded at 510 nm. The total flavonoid content was determined by using rutin as standard.

2.2.2.3 Total tannin content

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Various extracts of H. parviflora were analyzed for the presence of total tannins by the modified method of El Euch et al. [19] . Each extract (50 µL) was mixed with 1% of 100 mL vanillin in an ice bath. Then, the mixture was incubated at 37 ºC for 15 min and the absorbances of all the samples were recorded at 500 nm. The absorbance of the test sample was compared with the reference compound, Catechin to generate the calibration curve.

2.2.2.4 Estimation of total saponin content The total saponins contents of the tested extracts were analysis by respectively adding 50 μL of each plant extract to 250 μL of distilled water. After that, 250 μL of vanillin reagent

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and 2.5 mL of 72% H2SO4 were added to the mixture and incubated in a water bath at 60 °C for 10 min. Then, the mixture was cooled by using ice cold water and the absorbance was read at 544 nm.

2.3 Antioxidant activity

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DPPH radial scavenging ability, Superoxide radical scavenging potential, ABTS scavenging capacity, metal chelating ability and FRAP of all the five extracts of H. parviflora

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were estimated using standard protocols with ascorbic acid, rutin, α-Tocopherol, as standards, respectively [20]. Various concentrations of all the five extracts (50 to 1000

were calculated.

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2.4 Antidiabetic activity

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μg/mL) were studied for radical scavenging potentials. Finally, the percentages of inhibition

2.4.1 α- amylase inhibition assay

Antidiabetic activity of H. parviflora extracts were screened by α- amylase inhibitory

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assay. Various concentrations of leaf extracts were mixed with 500 μL of porcine pancreatic α-amylase and incubated at 25 °C for 10 min. Then, 500 μL of 1% starch solution was added

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to the solvent mixture and incubated at 37 °C for 5 min. Thereafter, 2.0 mL of 3, 5dinitrosalicylic acid was added to the incubated mixture. Then, the reaction was stopped by incubating in a boiling water bath at 100 °C for 15 min and cooling to room temperature. The reaction mixture was then diluted by adding 10 mL of distilled water in an ice bath, and the absorbance was measured at 540 nm. 2.4.2 α-glucosidase inhibition assay

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H. parviflora extracts were subjected to antidiabetic activity investigation by αglucosidase inhibitory assay. Each leaf extract was separately mixed with 100 μL αglucosidase in 0.2 M potassium phosphate buffer solution. Then, the mixture was incubated at 25 °C for 10 min. After incubation, 50 μL of 3 mM p-nitrophenyl and α-Dglucopyranoside-phosphate buffer solution were added. Again, the mixture was incubated at 37 °C for 30 min followed by the addition of 2 mL of sodium carbonate to the mixture. Then, the absorbance of the reaction mixture was determined at 405 nm. Acarbose was used as the positive control.

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3. Results and discussion Diabetes is a chronic metabolic disease, which is very common in developing countries. There are many medical plant extracts, which have been reported to have antihyperglycaemic activities and are also used in traditional medical practice. In recent times, screening of medical plants with antidiabetic activity has been increased as it is significant to

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discover novel cost effective drugs with less side effects for metabolic disorders such as diabetes [21]. The WHO has recommended people with diabetes to practice a healthy life

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style modification such as proper exercise and nutritious food habits as an efficient method of

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controlling the type II diabetes.

3.1 Phytochemical qualitative analysis

The evaluation of biologically active phytocompounds present in various parts of the

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medicinal plants is important to confirm about their medicinal values [22]. In the present study, leaves of H. parviflora were extracted using different polar solvents to confirm the various phytocompounds present. In the qualitative preliminary phytochemical analysis, the

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extracts such as HPLPE, HPLC, HPLEA, HPLE and HPLM (Table.1) exhibited the presence of phytocompounds such as alkaloids, flavonoids, tannins, steroids, triterpenoids, saponins

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and glycosides. Major secondary metabolites were present in the HPLM extract compared to the other extracts. Anthraquinones were absent in all the extracts of HPLPE, HPLEA, HPLC, HPLE and HPLM. The secondary metabolites with high medicinal values are used for various therapeutic purposes. Flavonoids found in the methanol extract possess a wide variety of biological activities, which include antimicrobial, antiangiogenic, anti-inflammatory, cytotoxic, antioxidant, antiviral and anticancer properties [23-25]. Terpenoids were present in all the extracts, which are reported to have anti-inflammatory, antimalarial, antiviral, 7

antibacterial activities and cause inhibition of cholesterol [26, 27]. Steroids were observed in the petroleum ether, ethyl acetate and chloroform extracts. They are responsible for reducing cholesterol levels and also have immune enhancing benefits [28]. Tannins present in the HPLE and HPLM extracts are known for several biological activities such as antiviral, antibacterial and anti-HIV [29] activities. The presence of alkaloids was reported in the HPLC, HPLE and HPLM extracts. Alkaloids have demonstrated various therapeutic activities such as antimicrobial [30], antiasthmatic, anti-inflammatory and antianaphylactic properties [31]. Ncube et al. reported that the yield of phytocompounds is mainly based on the extraction protocol and the solvent [32]. The solvents with different polarities are responsible

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for the separation of numerous phytocompounds, which has been reported in Chloroxylon swietenia [33]. The preliminary phytochemical results showed that the presence of major phytocompounds was based on the polarity of the solvents. Similar results were also reported in Chloroxylon swietenia and Turnera subulata [16, 33].

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3.2 Phytochemical quantification analysis

The quantification of phenolics, flavonoids, tannins and saponins was performed.

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Figure. 1 shows the phytochemical quantification in various extracts of H. parviflora. HPLM extract showed high yields viz., 18.9, 18.2, 0.9 and 39.9 mg/g of phenolics, flavonoids,

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tannins and saponins, respectively compared to the extracts. Phenolic compounds are powerful chain breaking antioxidants with very good scavenging abilities due to the presence of a hydroxyl group [34]. Similar phenolic and flavonoid contents were found in H. ponga

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[35] and other genus of the Diptercarpaceae family [36, 37]. Medicinal plants with rich phenolic contents are used in food industry because of their delayed lipid degradation property, which helps in improving the quality and nutritional value of food [38]. Flavonoids

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always owe a positive impact on human health; flavonoid derivatives have revealed a wide range of antiviral, anti-inflammatory, antibacterial, anallergic, and anticancer activities.

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Flavonoid compounds are highly efficient scavengers of high oxidizing molecules implicated in several diseases [39]. Tannins and saponins possess numerous

beneficial medical

properties such as antimicrobial, anti-inflammatory and antitumor activities [2].

3.3 Antioxidant activity In vitro antioxidant activities of the extracts have been evaluated by using DPPH, superoxide, FRAP, ABTS and metal chelating ferrous ion assays. DPPH, FRAP and ABTS

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assays are regularly used methods for the assessment of antioxidant activities of herbal extracts [40].

3.3.1 DPPH free radical scavenging activity The various solvent leaf extracts decreased DPPH colour from dark purple to light purple. The percentage of inhibition of each extract was noticed to be high (Fig. 2). The maximum percentages of inhibition of 58.12, 65.01, 47.34, 58.42, and 75.32 were reported in HPLPE, HPLC, HPLEA, HPLE and HPLM extracts, respectively. Among the various extracts, HPLM extract has showed the highest DPPH scavenging ability with IC50 value

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(100.12 µg/mL) as compared with the standard ascorbic acid (87.71%). Similar kind of results were observed for the Bucida buceras trunk, which could be attributed to their high phenolic contents [41].

3.3.2 Superoxide Radical scavenging activity

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The superoxide anion radical scavenging ability of each extract was examined with five different concentrations of solvent extracts. Superoxide radical scavenging activities of

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the extracts were measured by the reduction of nitroblue tetrazolium (NBT). NBT reduction was observed to be more for the HPLM extract than the other extracts. Similar kind of results

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were observed for Bucida buceras L [41]. The maximum percentage of superoxide scavenging by the HPLM extract was noticed as 71.38 at 1000 µg/mL. The potential IC50 value of the HPLM extract was 580.12 µg/mL, which was higher compared to the positive

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control ascorbic acid (280 µg/mL). Other extracts such as HPLPE, HPLC, HPLEA and HPLE showed the scavenging percentages of 55.27, 45.78, 53.36, and 59.25 % respectively (Fig. 3).

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3.3.3 FRAP Antioxidant power assay

Figure. 4 shows the FRAP scavenging activity of the H. parviflora extracts. Ion

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reduction capacity was noticed among the extracts in a dose dependent manner. Among the five extracts, the HPLM extract showed a good reducing absorbance (2.32) than the other extracts and the standard ascorbic acid [41].

3.3.4 ABTS Radical scavenging assay The antioxidant activities of the H. parviflora extracts were examined by using ABTS assay. ABTS radical scavenging ability of the HPLPE, HPLC, HPLEA, HPLE and HPLM extracts were evaluated using percentage inhibition of free radicals. The maximum ABTS 9

radical scavenging percentage was recorded in the HPLM extract as 71.28 percentage (Fig. 5) [42]. IC50 value of the HPLM extract was observed as 100.12 µg/mL and these results were more or less equal to ascorbic acid (91.2 µg/mL).

3.3.5 Metal chelating ability for ferrous ions The metal chelating abilities of the different extracts of H. parviflora were evaluated by the decrease in the formation of the ferrous- ferrozine complex. Among the extracts, the HPLEA demonstrated the highest activity of 78.71% at 250 µg/mL concentration (Fig. 6). The lowest IC50 value (180.35 µg/mL) was also observed in the HPLEA extract. The

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percentage inhibition of the standard ascorbic acid was 91.29 % and the IC50 value was 120.3 µg/mL [42].

In the present study, substantial differences in the free radical scavenging abilities of the various extracts of H. parviflora were observed based on the polarity of the solvent used. Among the five solvent extracts, the HPLM extract showed the maximum percentage of

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inhibition in the DPPH, superoxide and FRAP assays. In the ABTS and chelating ion assays, the HPLE and HPLEA extracts showed the most effective results. The scavenging activities

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increased with the increasing concentrations of the extracts and the results were comparable

3.4 Antidiabetic activity

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with the antioxidant activity of the leaf extracts of Hopea ponga [35].

Several remedial agents are available to treat diabetes, but they are expensive and also

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produced many side effects [21]. Conventional medicinal plants with antidiabetic activities can provide useful sources for the discovery of safer hypoglycaemic agents [43]. In the present research, various leaf extracts of H. parviflora were assessed for antidiabetic activity

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using standard in vitro techniques.

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3.4.1 α- amylase inhibitory activity α-amylase is one of the key enzymes in human body, which is responsible for starch

breakdown [44]. The α-amylase inhibition activities of various extracts of H. parviflora are provided in figure 6. Among the five extracts, the HPLM extract showed a comparable antidiabetic activity with the percentage of α-amylase inhibition being 76.87% at 500 µg/mL concentration (Fig. 7 and Table. 2) the IC50 value was higher (230.2 µg/mL) when compared with the standard acarbose (85.46 % inhibition with the IC50 value of 160.3 µg/mL). Almost similar activities were observed in the methanolic leaf extract of H. ponga. The second best 10

activity was observed in the HPLE extract with 68.03% inhibition at 500 µg/mL concentration and an IC50 value of 402.2 µg/mL. Plant based polyphenols play an important role in the inhibition of carbohydrate hydrolyzing enzymes such as α-amylase and αglucosidase [45]. The antidiabetic activity might have been due to the action of the extracts on α-amylase enzyme’s carbohydrate binding regions that catalyzed the hydrolysis of the internal α-1, 4 glucosidic starch linkage and other related polysaccharides, which have also been targeted for the postprandial hyperglycaemia suppression. Therefore, the present study has claimed that the natural polyphenols have α-amylase inhibitory activity and could be used as an effective therapy for the management of postprandial hyperglycaemia with minimal

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side effects [46]. 3.4.2 α- Glycosidase inhibitory activity

The results of α-glucosidase inhibitory actions and IC50 values of the various leaf extracts of H. parviflora at five different concentrations are summarized in Figure 8. and

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Table. 3. HPLM extract has shown the highest inhibitory activity compared with the standard acarbose. The HPLM extract has shown the maximum inhibitory activity of 82.26 ± 2.01% at

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500 µg/mL. The HPLE extract has shown the maximum inhibitory activity of 63.48 ± 0.62% at 400 µg/mL, similar kind of activity was observed in Orthosiphon stamineus [47]. For

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acarbose, the maximum inhibitory activity was found to be 88.11 ± 1.14% at 400 µg/mL [48]. This result exhibited that H. parviflora could be a potential source of natural supplement to replace commercially available pharmaceutical antidiabetic drugs in near future because it

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contains active phytocompounds, which act as α-glycosidase inhibitors. The HPLM extract showed the potential antioxidant and antidiabetic activities as compared with the commercial standards. These results indicated that the methanolic leaf

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extract of H. parviflora did possess strong antidiabetic and antioxidant potentials and would support traditional medicinal use for the treatment of diabetes mellitus and also would be a

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good source for natural antioxidants.

4. Conclusion

Our study showed that extracts from H. parviflora are easily accessible sources of phytocompounds possessing significant polyphenols and antioxidant activities, which attributes their effectiveness in the treatment of diseases such as diabetes mellitus. With reverence to the solvents used for extraction, methanol (HPLM) extract of H. parviflora contained high phenolic and flavonoid compounds, which is positively correlated with 11

stronger antioxidant and antidiabetic activities. These results indicated that the methanolic extract of H. parviflora can be used as a natural antioxidant and an antidiabetic agent.

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Figure Legends Figure 1. Comparative profiles of quantitative analysis of different solvent extracts of Hopea parviflora. Figure 2. Percentage of inhibition of DPPH free radical scavenging activities of various solvent extracts of Hopea parviflora.

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Figure 3. Percentage of inhibition of Superoxide scavenging activities of different solvent extracts of Hopea parviflora.

Figure 4. Percentage of inhibition of FRAP of different solvent extracts of Hopea parviflora. Figure 5. Percentage of inhibition of ABTS scavenging activities of different solvent extracts

-p

of Hopea parviflora.

Figure 6. Percentage of inhibition of Metal chelating abilities (ferrous ions) of different

re

solvent extracts of Hopea parviflora.

Figure 7. Comparative profiles of antidiabetic activities (Inhibition of α-amylase inhibitory activity) of various extracts of Hopea parviflora.

lP

Figure 8. Comparative profiles of antidiabetic activities (Inhibition of α- Glycosidase

Jo

ur

na

inhibitory activity) of various extracts of Hopea parviflora.

16

ro of

-p

re

lP

na

ur

Jo Figure 1.

17

ro of

-p

re

lP

na

ur

Jo Figure 2.

18

ro of

-p

re

lP

na

ur

Jo Figure 3.

19

ro of

-p

re

lP

na

ur

Jo Figure 4.

20

ro of

-p

re

lP

na

ur

Jo Figure 5.

21

ro of

-p

re

lP

na

ur

Jo Figure 6.

22

ro of

-p

re

lP

na

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Jo Figure 7.

23

ro of

-p

re

lP

na

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Jo Figure 8.

24

Table 1. Comparison of preliminary phytochemical qualitative screening of various fractions of Hopea parviflora S. No

Compound name

Various solvent extract HPLPE

HPLC

HPLEA

HPLE

HPLM

Alkaloids

-

+

-

+

+

2.

Flavonoids

-

-

-

-

+

3.

Tannins

-

-

-

+

+

4.

Steroids

+

+

+

-

-

5.

Triterpenoids

+

+

+

+

+

7.

Saponins

-

-

-

8.

Glycosides

+

+

+

9.

Gum & Mucilages

+

-

-

10.

Fixed oils

+

-

+

11.

Anthraquinones

-

-

-

-p

ro of

1.

-

-

-

+

-

-

+

+

-

-

Notes: (+) indicate presence of phytocompound and (-) absence of phytocompound.

re

HPLPE- Petroleum ether, HPLC- Chloroform, HPLEA- ethyl acetate, , HPLE- Ethanol,

lP

HPLM- Methanol.

Table 2. Inhibition of α-amylase activities of various extracts of Hopea parviflora.

Concentration

Jo

100 200

300

HPLPE

ur

(µg/mL)

na

α-amylase % of inhibition

Sample

8.11 ± 0.22 21.14

400

6.21 ± 11.02 ± 18.01 0.10

0.82

± 29.01

42.61±0.21

HPLM

A.

0.66

43.28±0.41

45.63±0.81 54.52±0.81

± 0.32

35.17 ± 38.71

± 0.60

0.50

38.51

45.05 ± 50.08

± 0.50

0.33

25

38.21±0.41

± 0.34

24.07 ± 26.07

± 0.45

0.73

HPLEA HPLE

acid

± 18.11

0.72 32.12

HPLC

58.57±0.14 65.32±0.95

± 0.65

± 0.20

69.21±0.20 78.52±1.31

500

58.31

± 51.38±

0.64

0.32

60.21 ± 68.03 0.43

76.87±0.22 85.46±1.14

± 0.12

HPLPE- Petroleum ether, HPLC- Chloroform, HPLEA- ethyl acetate, , HPLE- Ethanol, HPLM- Methanol. Each value in the table is represented as Mean ± SD. Values in the same column followed by different letter are significantly different (P<0.05).

α- Glycosidase % of inhibition

Sample Concentration

± 6.34

0.10

0.20

045 25.12

300

0.65 41.77 0.41

56.21

± 22.38

0.12

HPLE ± 14.61

0.29 ± 23.37

0.56

± 37.34

± 39.75

± 23.34

0.55

± 52.35

± 39.07

0.15

± 22.02±0.61 43.12±0.41

± 35.71±0.81 54.15±0.81

± 62.71±1.31 75.34±1.31

0.20

± 63.48

0.77

± 49.22±0.75 62.21±0.95

0.32

± 48.05

0.60

± 61.15

Acarbose

0.48

0.52

± 45.25

HPLM

0.45

0.76

0.77

ur

500

± 11.21

na

400

± 11.31

HPLEA

-p

7.21

18.34 200

HPLC

re

100

HPLPE

lP

(µg/mL)

ro of

Table 3. Inhibition of α- Glycosidase activities of various extracts of Hopea parviflora.

0.62

± 82.26

± 88.11±1.14

2.01

HPLPE- Petroleum ether, HPLC- Chloroform, HPLEA- ethyl acetate, HPLE- Ethanol,

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HPLM- Methanol.

Each value in the table is represented as Mean ± SD. Values in the same column followed by different letter are significantly different (P<0.05).

26