Asam gelugur Fruits
Garcinia atroviridis Griff. ex T.Anderson
Clusiaceae
Figure 1 : Garcinia atroviridis. (a) Tree; (b) leaves; (c) whole fruit; (d) abaxial and adaxial leaves; (e) cross section of fruit; (f) unripe fruit; (g) ripe fruit. (Photos courtesy of Ahmad Zariq Syafiq, NPRA, 2016)
DEFINITION
Asam gelugur fruits consist of the powder of dried deseeded fruits of Garcinia atroviridis Griff. ex. T.Anderson (Clusiaceae).
SYNONYM
Information and data have not been established.
VERNACULAR NAMES
Asam gelugur, assay gelugor, gelugor [ 1 ], assam gelugor, asam gelugo, asam keping (Malay) [ 2 ]; Suan zhu (Chinese) [ 3 ].
CHARACTER
| Colour | Brown |
| Odour | Characteristic |
| Taste | Sour |
IDENTIFICATION
Plant Morphology
G. atroviridis is a perennial tree grows to a height of more than 10–25 m. Trunk is long hardwood, fluted at the base with dull grey, cracked and fissured or smooth grey bark and drooping branches; inner bark produces a little transparent or yellow sap. Leaves coriaceous, pointed at the tip and abruptly tapered at the base, 15 x 4 cm – 25 x 7 cm, elongated oval, reddish young shoots when young and glossy dark green when mature. Inflorescence pedicellate, borne on the terminal of the twigs; calyx 4, yellow in colour, concave in shape and spreading orbicular; corolla 4, crimson, obovate and fleshy. Male flowers in short few-flowered racemes with numerous stamens in whorls round the pistillode on a fleshy receptacle. Female flowers solitary, large with an ovoid ribbed, 8–16 celled ovary; stigma, pileate, sub-tetragonal, convex in shape with deep red in colour; staminodes attached to an annulus. Fruits depressed globose, 6–10 cm diameter with broadly sunken concave apex, with 12–16 ribs and shallow grooves, green when young and turn to bright yellow when ripe. Seeds 1.5 cm long, surrounded by acidic, bright orange pulp [ 2 , 4 ].
Microscopy
Powdered material consists of parenchyma cells; fragment of reticulate and annular vessels; solitary crystal found isolated.
Figure 2 : Microscopic characters of Garcinia atroviridis driedfruit powder of 0.355 mm size. (a) Parenchyma cells; (b) reticulate and annular vessel; (c) solitary crystal. [Scale bars: a-e = 20 µm]
Colour Tests
Observed colour of solution after treatment with various reagents:
| HCl (conc.) | Brown |
| NaOH (5%) | Brown |
| FeCl3 (5%) | Yellow |
Thin Layer Chromatography (TLC)
Figure 3 : TLC profiles of xanthone (S) and methanol extract of Garcinia atroviridis dried fruit powder collected from different locations (L1, L2, L3) observed under (a) UV at 366 nm after spray with three prominent bands.
| Test Solutions | Weigh about 0.5 g of G. atroviridis dried fruit powder of 0.355 mm size in a screw cap sample bottle and add 5 mL of methanol. Sonicate for 15 min (without temperature setting). Filter the solution and use the filtrate as test solution. |
| Standard solution | Dissolve xanthone standard (CAS no.: 90-47-1) in methanol to produce 1 mg/mL solution. |
| Stationary Phase | HPTLC Glass Silica Gel 60 F254, 10 x 10 cm. |
| Mobile phase | Toluene : Ethyl acetate; (5 : 1) (v/v) |
| Application |
|
| Development distance | 8 cm |
| Drying | Air drying |
| Detection |
|
| Acceptance criteria |
|
High Performance Liquid Chromatography (HPLC)
| Test solution | Weigh about 20.0 mg of G. atroviridis dried fruit powder of 0.355 mm size in a 50 mL screw-capped conical flask and add 10 mL of purified water into the flask. Macerate the mixture in a water bath at 60ºC for 1 hr. Filter the mixture through a 0.45 µm syringe filter and inject the filtrate into the HPLC column. |
| Standard solution | Dissolve hydroxycitric acid [CAS no.: 6205-14-7] in purified water to produce 1 mg/mL solution. |
| Chromatographic system |
Detector: UV 200 nm Column: HILIC column (2.6 µm, 2.1 mm I.D x 100 mm) Column oven temperature: 30°C Flow rate: 0.2 mL/min Injection volume: 2 µL |
| Mobile Phase (gradient mode) | 6 mM phosphoric acid (in water) |
| Run time | 10 min |
| System suitability requirement |
Perform at least five replicate injections of the hydroxycitric acid standard solution (1 mg/mL). The requirements of the system suitability parameters are as follow:
|
| Acceptance criteria |
|
(a)
(b)
Figure 4 : Whole HPLC chromatogram of (a) hydroxycitric acid standard solution (1 mg/mL) at tr= 1.323 min and (b) water extract of Garcinia atroviridis dried fruit powder showing peak corresponding to hydroxycitric acid standard solution at tr= 1.347 min.
(a)
(b)
Figure 5 : HPLC chromatogram highlighting the elution region of hydroxycitric acid in (a) hydroxycitric acid standard solution (1 mg/mL) at tr= 1.323 min and (b) water extract of Garcinia atroviridis dried fruit powder showing peak corresponding to hydroxycitric acid standard solution at tr= 1.347 min.
PURITY TESTS
The purity tests are based on G. atroviridis dried fruit powder of 0.355 mm particle size.
| Foreign Matter |
| Not more than 2% |
| Ash Contents | |
| Total ash | Not more than 3% |
| Acid-insoluble ash | Not more than 1% |
| Loss on Drying |
| Not more than 10% |
| Extractive Values | |
| Water-soluble extracts | |
| Hot method | Not less than 50% |
| Cold method | Not less than 40% |
| Ethanol-soluble extracts | |
| Hot method | Not less than 40% |
| Cold method | Not less than 35% |
SAFETY TESTS
The safety tests are based on G. atroviridis dried fruit powder of 0.355 mm particle size.
| Heavy Metals | |
| Arsenic | Not more than 5.0 mg/kg |
| Mercury | Not more than 0.5 mg/kg |
| Lead | Not more than 10.0 mg/kg |
| Cadmium | Not more than 0.3 mg/kg |
| Microbial Limits | |
| Total bacterial count | Not more than 105 cfu/g |
| Total yeast and mould count | Not more than 104 cfu/g |
| Bile-tolerant gram negative | Not more than 104 cfu/g |
| Specific Pathogens | |
| Salmonella spp. | Absent in 25 g |
| Escherichia coli | Absent in 1 g |
| Staphylococcus aureus | Absent in 1 g |
| Pseudomonas aeruginosa | Absent in 1 g |
CHEMICAL CONSTITUENTS
Aqueous-methanol extract of G. atroviridis fruits was found to contain flavonoids (e.g. myricetin, quercetin and luteolin).
Methanol extract of G. atroviridis fruit was found to contain garcinia acid esters (e.g. 2-(butoxycarbonylmethyl)-3-butoxycarbonyl-2-hydroxy-3-propanolide and 1’,1”-dibutyl methyl hydroxycitrate) [ 5 , 6 , 7 ]
Essential oil of G. atroviridis was found to contain monoterpenes (e.g. linaloyl oxide, 1,4-cineol, p-cymene, p-cymenene and terpinolene), aldehydes (e.g. phenylacetaldehyde, benzaldehyde and nonanal), sesquiterpenes (e.g. β-panasinsene, (-)-β-caryophyllene, α-gurjunene, α-humulene, γ-selinene, β-selinene, α-selinene and δ-cadinene), polyphenolic (e.g. (E,E)-α-farnesene), sesquiterpenoid (e.g. γ-eudesmol), organic acids (e.g. tetradecanoic acid, pentadecanoic acid, hexadecanoic acid, (9Z,12Z)-9,12-octadecadienoic acid and (9Z)-9-octadecenoic acid), esters (e.g. ethyl tetradecanoate, (E,E)-farnesyl acetate, methyl hexadecanoate, 3-methylbutyl acetate, methyl 2-furoate, methyl (9Z,12Z)-9,12-octadecadienoate, methyl (9Z)-9-octadecenoate, ethyl (9Z,12Z)-9,12-octadecadienoate, ethyl (9Z)-9-octadecenoate and 2-phenylethyl acetate) and others ((E)-2-hexenal, (Z)-3-hexenol, hexanol, cis-herboxide, ocimenoyl oxide, cis-linalool oxide, 1,1-dimethyl-3-methylene-2-vinylcyclohexane, decanol, 1,1,6-trimethyl-1,2-dihydronaphthalene, 2-methylene-4,8,8-trimethyl-4-vinylbicyclo[5.2.0]nonane, (1R,2S,7S,9R)-2,6,10,10-tetramethyltricyclo[7.2.0.02,7]undec-5-ene, β-maaliene, caryophyllane-2,6-α-oxide, caryophyllane-2,6-β-oxide, (E)-4-(2’,6’,6’-trimethyl-1’,2’-epoxycyclohexyl)-3-penten-2-one, selina-4,11-diene, selina-3,7(11)-diene, β-caryophyllene alcohol, humulol, ginsenol, (1S,2S,5S,8S)-4,4,8-trimethyltricyclo[6.3.1.01,5]dodecan-2-ol, isophytol and trans-phytol ) [ 8 ].
MEDICINAL USES
Uses described in folk medicine, not supported by experimental or clinical data
Traditionally used after confinement by mixing the fruit with a lotion made with vinegar. The juice from the leaves is also used after childbirth. A decoction of leaves and roots is dropped into the ear canal for earache [ 9 ].
Biological and pharmacological activities supported by experimental data
Anti-atherosclerotic activity
Methanol extract of G. atroviridis fruit (50 mg/kg) was administered orally to diet induced atherosclerosis male Dunkin Hartley guinea pigs (700 – 1000 g) once daily for 60 days. The extract decreased the percentage of DNA damage (20%) compared to atherosclerotic treated group (28%) using alkaline comet assay [ 10 ].
Methanol extract of G. atroviridis fruit (50 mg/kg) was administered orally to cholesterol diet induced male Dunkin Hartley guinea pigs (700 – 1000 g) once daily for eight weeks. The extract reduced the total cholesterol (3.61 ± 0.97 mmol/L) compared to cholesterol diet group (4.57 ± 1.90 mmol/L) [ 11 ].
Antihyperlipidemic activity
Aqueous extract of G. atroviridis rind (1000 mg/kg) was administered orally to polazamer 407-induced acute hyperlipidemic male Sprague Dawley rats (aged between 12 to 16 weeks; 180 – 250 g) once daily for three days. The extract reduced the low-density lipoprotein (LDL) (3.15 ± 0.34 mmol/L) and very-low-density lipoprotein (VLDL) (2.15 ± 0.46 mmol/L) compared to hyperlipidemic control group (LDL: 4.24 ± 0.17 mmol/L; VLDL: 4.91 ± 1.17 mmol/L). The atherogenic index (AI) of the extract showed significant (p < 0.05) reduction (10.22 ± 1.87) compared to hyperlipidemic control group (AI: 20.62 ± 2.64 mmol/L) [ 17 ].
Antibacterial activity
Total volatiles compound from G. atroviridis fresh fruits showed antibacterial activity on Gram positive bacteria with minimum inhibitory concentration (MIC) values of 4.8 µg/mL for Bacillus subtilis and 4.8 µg/mL for Staphylococcus aureus compared to gentamicin sulfate as positive control (MIC values of 0.4 µg/mL for B. subtilis and 0.1 µg/mL for S. aureus) using microdilution assay [ 12 ].
Antifungal activity
Methanol extract of G. atroviridis fruits showed antifungal activity with minimum inhibitory dose (MID) value of 100 µg/spot on Cladosporium herbarum [ 13 ].
1’, 1”-dibutylmethyl hydroxycitrate and 2-(butoxycarbonylmethyl)-3-butoxycarbonyl-2-hydroxy-3-propanolide isolated from methanol extract of fruits of G. atroviridis inhibited growth of Cladosporium herbarum with MID values of 0.4 μg/spot and 0.8 μg/spot, respectively compared to cycloheximide (MID = 0.5 μg/spot) [ 6 ].
Anti-inflammatory activity
Total volatiles compound from G. atroviridis fresh fruits showed anti-inflammatory activity on ovine cyclooxygenase-1 (COX-1) with inhibition concentration at 50% (IC50) value of 4.64 ± 0.6 µg/mL and human recombinant COX-2 (IC50 value = 2.53 ± 0.8 µg/mL) compared to positive control indomethacin (COX-1: 3.31 ± 0.3 µg/mL; COX-2: 4.72 ± 0.6 µg/mL) [ 12 ].
Antioxidant activity
2-(butoxycarbonylmethyl)-3-butoxycarbonyl-2-hydroxy-3-propanolide isolated from G. atroviridis fruits showed antioxidant activity by using ferric thiocyanate (FTC) assay (11%) and thiobarbituric acid (TBA) assay (37%). The compound also showed antioxidant activity using radical scavenging assay (3.4%) compared to ascorbic acid (88.2%) and butylated hydroxytoluene (BHT) (80.5%) [ 14 ].
1′, 1”-dibutylmethyl hydroxycitrate isolated from G. atroviridis fruits showed antioxidant activity by using FTC assay (36%) and TBA assay (53%). The compound also showed antioxidant activity using radical scavenging assay (5.5%) compared to ascorbic acid (88.2%) and butylated hydroxytoluene (BHT) (80.5%) [ 14 ].
Aqueous and methanol extract of G. atroviridis fruits showed antioxidant activity with total phenolic contents of (32.53 ± 0.56 mg gallic acid equivalent(GAE)/100 g and 62.34 ± 3.07 mg GAE/100 g, respectively) and Fe2+ reducing ability of (434.17 ± 28.34 µM/g and 624.17 ± 12.58 µM/g, respectively) using ferric reducing antioxidant power (FRAP) assay [ 18 ].
Antitumour activity
2-butoxycarbonylmethyl-3-butoxycarbonyl-2-hydroxy-3-propanolide isolated from G. atroviridis fruitsshowed antitumour activity by inhibiting Epstein-Barr virus activation on Raji cells (IC50 value = 70 µM) [ 14 ].
Methanol extract of G. atroviridis fruits inhibit the tumour-promoter-induced Epstein Barr virus in Raji cells by the by 67.5% compared to untreated cells (control) (0.0%) [ 13 ].
Cytotoxic activity
Methanol extract of G. atroviridis fruits showed no cytotoxic effect on human T-lymphoblastic leukaemia cell (CEM-SS) with cytotoxic dose (CD50) value of > 30 µg/mL after 72 hours of incubation using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay [ 13 ].
Clinical studies
Reduction of adipose tissue and body weight
A randomized double-blind placebo controlled trial showed that 25 obese Thai women who ingested (-)-Hydroxycitric acid (HCA) sachet containing 1.15 g G. atroviridis three times daily for a duration of 8 weeks significantly (p < 0.05) reduced triceps skinfold, biceps skinfold, subscapular skinfold, suprailiac skinfold, upper arm circumference, percentage of the body fat and body fat weight (kg) compared to placebo group [ 15 ].
SAFETY INFORMATION
Preclinical studies (Toxicology studies)
14-days acute oral toxicity
Oral single dose acute toxicity study on female Sprague Dawley rats (aged between 8 to 12 weeks old) using aqueous extract of G. atroviridis fruits showed no toxic effect on the parameters observed including behaviors, body weight, food and water intakes. All rats were observed for 14 days prior to necropsy. No death was found throughout the study period. Necropsy revealed no significant abnormality. Approximate lethal dose (LD50) is more than 2,000 mg/kg body weight [16 ].
Others (Adverse reaction, contraindication, side effect, warning, precaution)
Information and data have not been established.
DOSAGE
Information and data have not been established.
STORAGE
Store below 30°C. Protect from light and moisture.
REFERENCES
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- Pangsuban S, Bamroongrugsa N, Kanchanapoom K and Nualsri C. An evaluation of the sexual system of Garcinia atroviridis (Clusiaceae), based on reproductive features. Songklanakarin Journal of Science and Technology 2007;29(6):1457–1468.
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- Amran AA, Zaiton Z, Faizah O, Morat P. Effects of Garcinia atroviridis on serum profiles and atherosclerotic lesions in the aorta of guinea pigs fed a high cholesterol diet. Singapore Medical Journal. 2009;50(3):295-299.
- Tan WN, Wong KC, Khairuddean M, Eldeen IM, Asmawi MZ, Sulaiman B. Volatile constituents of the fruit of Garcinia atroviridis and their antibacterial and anti-inflammatory activities. Flavour and Fragrance Journal. 2013;28:2-9.
- Mackeen MM, Ali AM, Lajis NH, Kawazu K, Hassan Z, Amran M, Habsah M, Mooi LY, Mohamed SM. Antimicrobial, antioxidant, antitumour-promoting and cytotoxic activities of different plant part extracts of Garcinia atroviridis Griff. ex T. Anders. Journal of Ethnopharmacology. 2000;72:395-402.
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- Roongpisuthipong C, Kantawan R, Roongpisuthipong W. Reduction of adipose tissue and body weight: effect of water soluble calcium hydroxycitrate in Garcinia atroviridis on the short term treatment of obese women in Thailand. Asia Pacific Journal of Clinical Nutrition. 2007;16(1):25-29.
- Teh BP, Hamzah NF, Rosli SNS, Yahaya MAF, Zakiah I, Murizal Z. Acute oral toxicity study of selected Malaysian medicinal herbs on Sprague Dawley rats. Institute for Medical Research, Ministry of Health; 2012. Report No. HMRC 11-045/01/GA/F/A.
- Al-Mansoub MA, Asmawi MZ, Murugaiyah V. Effect of extraction solvents and plant parts used on the antihyperlipidemic and antioxidant effects of Garcinia atroviridis: a comparative study. Journal of the Science of Food and Agriculture. 2014;94(8):1552-1558.
- Rabeta MS, Nur Faraniza R. Total phenolic content and ferric reducing antioxidant power of the leaves and fruits of Garcinia atroviridis and Cynometra cauliflora. International Food Research Journal. 2013;20(4):1691-1696.
