Journal of Herbs, Spices & Medicinal Plants Chemical

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Chemical Composition of Garcinia xanthochymus Seeds, Seed Oil, and Evaluation of its Antimicrobial and Antioxidant Activity S. H. Manohar N. Murthy

a b

, P. M. Naik

a c

a

a

, L. M. Patil , S. I. Karikatti & H.

a

a

Department of Botany, Plant Biotechnology Laboratory , Karnatak University , Dharwad , India b

Department of Biotechnology , Jain University , Bangalore , India

c

Department of Genetics and Plant Breeding , University of Agricultural Sciences , Dharwad , India Published online: 14 Feb 2014.

To cite this article: S. H. Manohar , P. M. Naik , L. M. Patil , S. I. Karikatti & H. N. Murthy (2014) Chemical Composition of Garcinia xanthochymus Seeds, Seed Oil, and Evaluation of its Antimicrobial and Antioxidant Activity, Journal of Herbs, Spices & Medicinal Plants, 20:2, 148-155, DOI: 10.1080/10496475.2013.847886 To link to this article: http://dx.doi.org/10.1080/10496475.2013.847886

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Journal of Herbs, Spices & Medicinal Plants, 20:148–155, 2014 Copyright © Taylor & Francis Group, LLC ISSN: 1049-6475 print/1540-3580 online DOI: 10.1080/10496475.2013.847886


Chemical Composition of Garcinia xanthochymus Seeds, Seed Oil, and Evaluation of its Antimicrobial and Antioxidant Activity S. H. MANOHAR,1,2 P. M. NAIK,1,3 L. M. PATIL,1 S. I. KARIKATTI,1 and H. N. MURTHY1 1

Department of Botany, Plant Biotechnology Laboratory, Karnatak University, Dharwad, India 2 Department of Biotechnology, Jain University, Bangalore, India 3 Department of Genetics and Plant Breeding, University of Agricultural Sciences, Dharwad, India

Seeds of Garcinia xanthochymus contain (in g.100−1 ) 2.35 ash, 6.93 protein, 45.22 carbohydrate, 12.3 crude fiber, and saturated and unsaturated fatty acids at 34.17% and 65.79%, respectively. Seed oil was composed of nine major fatty acids including myristic acid (0.11%), palmitic acid (32.96%), stearic acid (0.96%), palmitoleic acid (17.65%), oleic acid (45.87%), linoleic acid (1.93%), linolenic acid (0.34%), arachidic acid (0.07%), and behenic acid (0.07%). The oil demonstrated antimicrobial activity against gram-positive bacteria. Antioxidant activities of oil were evaluated using α,α-diphenyl-β-picrylhdrazyl reduction activities, and it was higher than that of ascorbic acid and butylated hydroxyanisole (IC50 = 120 μg.mL−1 ). KEYWORDS Unsaturated fatty acid, saturated fatty acid

INTRODUCTION The continued increase in world population and the ever-increasing demand for edible oil have resulted in increase in the prices of oils. In the search for new sources of novel oils, a large number of plants have been surveyed, and Received February 6, 2013. Address correspondence to H. N. Murthy, Department of Botany, Plant Biotechnology Laboratory, Karnatak University, Pavate Nagar, Dharwad 580 003, India. E-mail: nmurthy60@ yahoo.co.in 148


Chemical Composition of Garcinia xanthochymus Seeds

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some of the most promising species have been cultivated as new oil crops (12). The study of nutritive value, methods of production, preservation, and utilization is important for their effective uses. There are hundreds of species that have received much less attention from the scientific community than the annual crops (9). Garcinia xanthochymus is popularly known as Yellow mangosteen and is native to India and Myanmar. The plant is known to posses several important phytochemicals that have been demonstrated to possess antibacterial (6), anti-inflammatory (18), and antioxidant (22) activities. The bark is used as astringent (14). Traditionally, the fruits are used as antiscorbutic, cooling, digestive, emollient, demulcent, cholagogue, and sherbet made from dried fruit is used in biliousness. Garcinia mangostana and G. indica yield edible and industrial oil (1,13). The aims of the study are to analyze the chemical composition of G. xanthyochums seeds and seed oil and evaluate its antimicrobial and antioxidant activities.

MATERIALS AND METHODS Plant Material and Sample Preparation G. xanthochymus fruits were collected from Bakkala garden in Sirsi, Karnataka, India. The seeds were removed from the fruits and washed with water. Approximately 100 g of seed was dried overnight at 80◦ C and ground in a coffee grinder. The oil was extracted for 8 h with petroleum ether (b.p. 40–60◦ C) as a solvent in a Soxhlet extractor. The solvent was removed completely under vacuum, and the oil thus obtained was used for further analysis. Extraction was performed in triplicates.

Proximate Analysis of Seed Proximate analysis of moisture content, crude fat, crude protein, crude fiber, carbohydrate, and ash was done (2).

Physico-Chemical Properties of Seed Oil Oil from the seed was subjected to physical characterization. The color and state of the oil at room temperature were noted by visual inspection, and specific gravity was determined (3). The refractive index of the oil at room temperature was estimated using the Abbe refractometer (4). Free fatty acids (FFA), iodine value (IV), saponification number (SN), peroxide value (PV), and unsaponifiable matter were measured (5). The analysis was performed for the freshly extracted oils. The samples were analyzed in triplicates and expressed as mean ± SE.

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Fatty Acid Analysis


Fatty acid methyl esters of G. xanthochymous seed oil were analyzed on a Chemito G. C. 8610 gas chromatograph equipped with flame ionization detector and capillary column B P × 70 (50 m × 0.32 mm × 0.25 µm films). The detector temperature was 260◦ C with flow rate of 0.3 mL.min−1 . The injector temperature was set at 240◦ C, and nitrogen (purity 99.95 %) was used as the carrier gas. Identification of the peaks was performed by comparing retention times with standards.

Antimicrobial Analysis Antimicrobial activity of G. xanthochymous seed oil was tested toward 15 microorganisms by the disc diffusion method according to the National Committee for Clinical Laboratory Standards Guidelines (16). Six grampositive bacteria— Enterococcus faecalis ATCC 29212; Staphylococcus aureus ATCC 29213; Vancomycin Resistant Enterococcus (VRE) ATCC 51299; Bacillus subtilis SDMC 025; Micrococcus spp. SDMC 016; and Staphylococcus epidermidis SDMC 097—and six gram-negative bacteria—Escherichia coli ATCC 25922; Pseudomonas aeruginosa ATCC 27853; Proteus mirabilis SDMC 042; Salmonella paratyphi-A SDMC 014; Salmonella paratyphi-B SDMC 011; and Providencia alcalifaciens SDMC 056—were used. Aspergillus niger SDMC 052, Penicillium notatum SDMC 064, and Candida albicans SDMC 033 were the three fungi used for the study. The microorganisms used for the analysis were obtained from American Type Culture Collection and cultures maintained at Sri Dharmasthala Manjunatheswar Medical College, Dharwad, Karnataka, India. The minimal inhibitory concentrations of the oil were determined by micro-dilution assay. The oil was two-fold serially diluted with DMSO, which contained 0.125–16 µg.µL−1 of oil (15).

Antioxidant Activity: Free Radical-Scavenging Capacity The abilities of the oil to scavenge α,α-diphenyl-β-picrylhdrazyl (DPPH) free radicals were measured (20). Ascorbic acid (ASC) and butylated hydroxyanisole (BHA) were also monitored for radical-scavenging activity. Inhibition of free radical by DPPH in percent was calculated as scavenging DPPH (%) = 100 × (Ablank - Asample/A blank), where Ablank is the absorbance of the control reaction (containing all reagents except the oil) and Asample is the absorbance of the sample. The percentage of scavenged DPPH was plotted versus the concentration of antioxidants and the concentration of antioxidant required to obtain 50% inhibition (50% inhibition concentration or IC50 ).


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RESULTS Moisture, ash, crude protein, carbohydrate, and crude fiber content of the seed were (in g.100 g−1 ) 11.96 ± 0.52, 2.35 ± 0.29, 6.93 ± 0.64, 45.22 ± 1.20, and 12.3 ± 1.66, respectively. The seed oil was golden-orange in color and was consistently liquid at room temperatures (25.0◦ ± 2.0◦ C; Table 1). The iodine value 94.86 ± 0.29 mg.100 g−1 indicated this oil was non-drying and high in unsaturated fatty acids. Fatty acid profile of G. xanthochymous seed oil was composed of nine major fatty acids: myristic acid, palmitic acid, stearic acid, palmitoleic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid, and behenic (Figure 1, Table 2). The concentration of saturated fatty acids (SFA) and unsaturated fatty acids (UFA) were 34.17% and 65.79%, respectively. The most prevalent UFA were oleic acid (45.8 g.100g−1 ) and palmitoleic acid (17.6 g.100g−1 ), and the most prevalent SFA was palmitic acid (32.96 g.100g−1 ). The oil showed inhibition against the tested gram-positive bacteria (S. aureus, B. subtilis, Micrococcus sps, and S. epidermidis) and no activity against the tested gram-negative bacteria and fungus (Table 3). The oil reduced the stable radical DPPH to the yellow-colored DPPH-H with an IC50 value of 120 µg.mL−1 . Ascorbic acid (ASC) and butylated hydroxyanisole (BHA) as two positive controls exhibited high antioxidant activity with IC50 values 4.25 µg.mL−1 and 5.66 µg.mL−1 , respectively.

DISCUSSION Although the moisture content of G. xanthochymus was less than G. mangostana (13.08 ± 1.99 g.100 g−1 ), the ash content was higher than that of G. mangostana (1). The crude protein content is comparable with 6.57 g.100g−1 TABLE 1 Physicochemical Characteristics of the Seed Oil of Garcinia xanthochymous∗ Component Acid value (mg NaOH/g oil) Saponification number (mg KOH/g oil) Unsaponifiable matter Iodine value (mg/100 g) FFA (%) as oleic acid Peroxide value (mg/g oil) Ester value (mg/KOH) State at RT Color Specific gravity Refractive index at RT FFA (%), free fatty acid (%); RT, room temperature. ∗ Values are means ± standard error of triplicate determinations.

Garcinia xanthocymus oil 5.43 ± 0.63 169.21 ± 1.12 1.20 ± 0.06 94.86 ± 0.29 2.73 ± 0.32 13.9 ± 1.46 141.1 ± 2.69 Liquid Golden-orange 0.971 ± 0.01 1.488


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FIGURE 1 Gas chromatogram of fatty acids profile of Garcinia xanthochymous seed oil. TABLE 2 Fatty Acid Composition of Garcinia xanthochymous Seed Oil Fatty Acid

Values

C14:0 Myristic C16:0 Palmitic C18:0 Stearic C16:1 Palmitoleic C18:1 Oleic C18:2 Linoleic C18:3 Linolenic C20:0 Arachidic C22:0 Behenic Total saturates Total unsaturates

0.11 32.96 0.96 17.65 45.87 1.93 0.34 0.07 0.07 34.17 65.79

for G. mangostana (1). The seeds contained 31.7 ± 1.74 g.100 g−1 crude fat, which is lower than 45.5 g.100 g−1 reported for G. indica (13) but higher than 21.18 ± 6.18 g.100 g−1 reported for G. mangostana seeds (1). The carbohydrate and crude fiber contents were similar to that G. mangostana (43.5 ± 2.09 g.100 g−1 and 13.7 ± 0.89 g.100 g−1 ). The seeds are rich in carbohydrates and proteins, could serve as source of roughage, and can be used in animal feeds. Refractive index of the oil was 1.488, comparable with that of G. mangostana (1). The total acidity, expressed as acid value was 5.43 ± 0.63 mg


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TABLE 3 Antimicrobial Activity and Minimum Inhibitory Concentration of Garcinia xanthochymous Seed Oil


Organism

Garcinia xanthochymous oil zone of inhibition (mm) and MIC (µg.mL−1 )

S. aureus ATCC 29213 B. subtilis SDMC 025 Micrococcus SDMC 016 S. epidermidis SDMC 097 E. faecalis ATCC 29212 V.R.E ATCC 51299 E. coli ATCC 25922 P. aeruginosa ATCC 27853 S. paratyphi A SDMC 014 S. paratyphi B SDMC 011 P. mirabilis SDMC 042 Pr. Alcalifaciens SDMC 056 A. niger SDMC 052 P. notatum SDMC 064 C. albicans SDMC 033

18 ≤ 15 ≤ 18 ≤ 16 ≤ — — — — — — — — — — —

4 8 4 4

Standard Zone of inhibition (mm) and MIC (µg.mL−1 ) 29 23 32 20 30 21 24 15 12 13 24 11 13 09 10

≤ ≤ ≤ ≤ ≤ ≤ ≤ ≤ ≤ ≤ ≤ ≤ ≤ ≤ ≤

0.005 0.010 0.005 0.005 0.010 0.010 0.005 0.010 0.020 0.020 0.010 0.020 0.040 0.160 0.160

MIC, minimum inhibitory concentration.

NaOH/ g and was within the allowable limits of edible oils (10). The saponification value was 169.21 ± 1.12 mg and KOH/g was comparable to common vegetable oils (19). Free fatty acid (FFA) content of G. xanthochymous at 2.73 ± 0.32% was similar to other edible oils (7) with long shelf life. The peroxide value of the oil was 13.9 ± 1.46 g.100 g−1 , suggesting that it can be stored for a long period without deterioration; however, this value is higher than the seed oil of G. mangostana (3.27 ± 0.12 g.100 g−1 ). The concentration of SFA and UFA were varied with respect to G. mangostana (1), which was SFA, 59.6% and UFA, 35.3%. Free radicals are believed to be involved in bacterial and parasitic infections, lung damage, inflammation, reperfusion injury, cardiovascular disorders, atherosclerosis, aging, and neoplastic diseases (8). In biochemical systems, H2 O2 generates extremely reactive hydroxyl radicals in the presence of certain transition metal ions (e.g., iron and copper) or by ultraviolet photolysis (21). Hydroxyl radicals can attack DNA molecules, cause lipid peroxidation (11), tissue damage, protein denaturation, and glutathione depletion (17). The oil obtained from G. xanthochymus seeds has shown significant antioxidant activity and hence of significant health value.

FUNDING This work was partially financed by the University Grants Commission under the Special Assistance program, New Delhi, India. Authors are

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thankful to the Department of Biotechnology (DBT-KUD-IPLS program BT/PR14555/INF/22/126/2010), University Grants Commission [Project No. F. No. 41-423/2012 (SR)], New Delhi and Department of Atomic Energy (BRNS project No. 2013/35/BRNS/20), Mumbai for financial assistance.


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