Gelenggang Leaves
Senna alata (L.) Roxb
Fabaceae
Figure 1 : S. alata. (a) Whole plant; (b) leaves; (c) flowers; (d) seeds. (Photos courtesy of FRIM, 2012)
DEFINITION
Gelenggang leaves consist of dried leaves of S. alata (L.) Roxb. (Fabaceae)
SYNONYM
VERNACULAR NAMES
Seven golden candlesticks, ringworm bush (English), daun kurap, gelenggang, ludanggan (Malay), chi jia jue ming (Chinese), seemaiagathi (Tamil) [ 1 , 3 ].
CHARACTER
| Colour | Dark green |
| Odour | Unpleasant |
| Taste | No taste |
IDENTIFICATION
Plant Morphology
A shrub reaching 1-2(-5) m high, branches greenish, thick and pubescent. Leaves pinnate with a long rachis about 30-60 cm long; each pinna has about 8-20 pairs of leaflets; petiole 2-3 cm long; leaflets 5-15 cm x 3-7 cm, oblong-elliptical with rounded corners or obtuse at the ends and glabrous. Inflorescence is an axillary raceme, robust, dense, 20-50 cm long, large, very erect and very easily seen even from a distance. Flowers with oblong sepals, 10-20 mm x 6-7 mm, orange-yellow; petals ovate-orbicular, 16-24 mm x 10-15 mm, bright yellow; 2 large stamens with stout filaments of 4 mm long and anthers about 12-13 mm long; 4 small stamens with filaments of 2 mm long and anthers about 4-5 mm long; 3-4 staminodes; ovary puberulous, sessile, with many (up to 58) ovules; style filiform; stigma small. Fruit tetragonal, 10-15 cm x 1.5-8 cm, winged 4-8 mm, black, glabrous, up to 50-seeded. Seeds quadrangular, flattened, 7-8 mm x 5-8 mm, shiny [ 2 , 4 ].
Microscopy
Microscopic characters of S. alata leaf powder consist of epidermis adaxial cells with straight to wavy anticlinal wall, epidermis abaxial cells with sinuous anticlinal wall and attached with stomata; the fragment of vessel cells, tracheid and solitary calcium oxalate crystals are present; the less abundant simple unicellular trichomes with very rough surface are also present; the abundant starch granules are mostly simple, oblong to sub-rectangular in shape.
Figure 2 : Microscopic characters of S. alata leaves powder. (a) Epidermis adaxial cells (magnification 20x); (b) epidermis abaxial cells attached with stomata (magnification 20x); (c) vessel cells (magnification 100x); (d) tracheid (magnification 10x); (e) solitary crystal (magnification 20x); (f) simple unicellular trichome (magnification 100x); (g) starch granules (magnification 100x). [Scale bars: a, b, e = 20 µm; c, f, g = 5 µm; d = 50 µm]
Colour Tests
Observed colour of solution after treatment with various reagents :
| 5% NaOH | Yellow |
| 25% NH4OH | Brown |
Thin Layer Chromatography (TLC)
Figure 3 : TLC profiles of kaempferol-3-O-gentiobioside (S) and methanol extracts of S. alata dried leaves powder (1, 2 & 3) observed under (a) UV 254 nm, (b) UV 254 nm after spraying with natural product reagent and (c) UV 366 nm after spraying with natural product reagent.
| Test Solutions | Weigh about 1.0 g of S. alata dried leaves powder in a 50 mL screw-capped conical flask and add 10 mL of methanol and sonicate for 15 minutes. Filter the solution through filter paper. Use the filtrate as test solution. |
| Standard solution | Dissolve 5.0 mg of kaempferol-3-O-gentiobioside standard in 5 mL methanol to produce 1000 µg/mL solution. |
| Stationary Phase | HPTLC Glass Silica Gel 60 F254, 10 x 10 cm. |
| Mobile phase | Methanol: ethyl acetate: water: formic acid (10:30:5:1) (v/v) |
| Application |
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| Development distance | 8 cm |
| Drying | Air drying |
| Detection |
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High Performance Liquid Chromatography (HPLC)
| Test solution | Extract about 0.5 g of S. alata dried leaves powder with 3 mL of methanol in a 7 mL screw-capped bottle. Sonicate the mixture for 2 hours at room temperature. Filter the mixture solution through a 0.45 µm syringe filter and inject the filtrate into the HPLC column. | ||||||||||||||||||
| Standard solution | Dissolve a quantity of kaempferol-3-O-gentiobioside in methanol to produce a solution containing 1 mg/mL. | ||||||||||||||||||
| Chromatographic system |
Detector: UV 350 nm Column: C18 (5 µm, 4.6 mm I.D x 250 mm) Column oven temperature: Ambient Flow rate: 1.0 mL/min Injection volume: 5 µL |
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| Mobile Phase (gradient mode) |
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| System suitability requirement |
Perform at least five replicate injections of the standard solutions (1.0 mg/mL). The requirements of the system suitability parameters are as follow:
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| Acceptance criteria |
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(a)
(b)
Figure 4 : HPLC chromatogram (a) refers to full and (b) refers to zoom chromatograms) of kaempferol-3-O-gentiobioside standard solution (1.0 mg/mL) at tr = 18.24 min.
(a)(i)
(a)(ii)
(b)(i)
(b)(ii)
(c)(i)
(c)(ii)
Figure 5 : HPLC chromatograms ((i) refers to full and (ii) refers to zoom chromatograms) of methanol extracts of S. alata leaves from different locations ((a) refers to location 1, (b) refers to location 2 and (c) refers to location 3).
Figure 6 : UV spectra of kaempferol-3-O-gentiobioside standard solution (1.0 mg/mL) and methanol extracts of S. alata leaves.
PURITY TESTS
| Foreign Matter |
| Not more than 2% |
| Ash Contents | |
| Total ash | Not more than 10% |
| Acid-insoluble ash | Not more than 1% |
| Loss on Drying |
| Not more than 15% |
| Extractive Values | |
| Water-soluble extracts | |
| Hot method | Not less than 20% |
| Cold method | Not less than 18% |
| Ethanol-soluble extracts | |
| Hot method | Not less than 16% |
| Cold method | Not less than 10% |
SAFETY TESTS
| 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
Methanolic extract of the leaves has been found to contain flavonoids (e.g. kaempferol, kaempferol-3-O-gentiobioside and kaempferol-3-O-β-d-glucopyranoside), anthraquinones (e.g. aloe-emodin, rhein methyl ester and danthron), diterpene (e.g. phytol) and minor compound (e.g. 4-hydroxybenzoic acid) [ 5 , 6 ].
Ethanolic extract of the leaves contains several minor compounds such as p-hydroxybenzoic acid and adenine [ 7 , 8 ].
Butanol extract of the leaves also contains flavonoids (e.g. kaempferol, kaempferol-3-O-β-d-glucopyranoside, kaempferol-3-O-gentiobioside) and anthraquinones (e.g. rhein and aloe-emodin) [ 9 , 10 ].
Chloroform extract of the leaves contain santhraquinones (e.g. aloe-emodin and rhein) and steroidal compounds (e.g. stigmasterol and sitosterol) [ 11 , 12 ].
Hexane extract of leaves has been found to contain cassiaindoline [ 13 ].
Essential oils of the leaves contain mainly monoterpenes (e.g. linalool, borneol, α-terpineol, 1,8-cineole and limonene), sesquiterpenes (e.g. β-caryophyllene, caryophyllene oxide, germacrene and α-selinene) and lipid (e.g. pentadecanal) [ 14 , 15 ].
MEDICINAL USES
Uses described in folk medicine, not supported by experimental or clinical data
Traditionally, the Malays and the Chinese rubbed the pounded leaves mix with lime on the skin as a remedy for ringworm and other cutaneous diseases. A decoction of the leaves also may be taken as a purgative [ 1 ].
Biological and pharmacological activities supported by experimental data
Antimicrobial activity
Methanol extract of S. alata leaves (7.8-500 µg/mL) inhibited the growth of Bacillus subtilis with minimum inhibitory concentration (MIC = 7.8 µg/mL), Pseudomonas aeruginosa (MIC = 125 µg/mL), Escherichia coli (MIC = 125 µg/mL), Staphylococcus aureus (MIC = 125 µg/mL) and Proteus vulgaris (MIC = 125 µg/mL) with inhibition zone ranging from 5 to 11 mm compared to gentamicin (5 mm) using agar diffusion assay [ 16 ].
Methanol extract of S. alata leaves (3-100 µg/mL) inhibited the growth of E. coli (MIC = 15 µg/mL), Proteus mirabilis (MIC = 20 µg/mL), P. aeruginosa (MIC = 20 µg/mL), Salmonella typhi (MIC = 15 µg/mL), Shigella flexnerri (MIC = 15 µg/mL), S. aureus (MIC = 12 µg/mL) and Streptococcus pyogenes (MIC = 12 µg/mL) with inhibition zones ranging from 6 to12 mm compared to ciprofloxacin (inhibition zones=12-28 mm) and streptomycin (inhibition zones = 6-22 mm) using agar diffusion assay [ 16 , 17 ].
Chloroform extract of S. alata leaves (7.8-500 µg/mL) inhibited the growth of B. subtilis (MIC = 62.5 µg/mL), P. aeruginosa (MIC = 250 µg/mL), E. coli (MIC = 250 µg/mL) and S. aureus (MIC = 62.5 µg/mL) with inhibition zone ranging from 4 to 8 mm compared to gentamicin (5 mm) using agar diffusion assay [ 16 ].
Aqueous extract of S. alata leaves (40-160 mg/mL) inhibited the growth of S. aureus (MIC = 25.12 mg/mL), E. coli (MIC = 3.55 mg/mL) and S. typhi (MIC = 22.39 mg/mL) with inhibition zones ranging from 11.60 to 37.00 mm compared to rifampicin (23-27 mm) using agar diffusion assay [ 18 ].
Ethanol (95%) extract of S. alata leaves (20-160 mg/mL) inhibited the growth of S. aureus (MIC = 0.00 mg/mL), E. coli (MIC = 3.55 mg/mL) and S. typhi (MIC = 1.41 mg/mL) with inhibition zones ranging from 15.80 to 29.40 mm compared to rifampicin (23 – 27 mm) using agar diffusion assay [ 18 ].
Methanol extract of S. alata leaves (3-100 µg/mL) inhibited the growth of Aspergillus flavus (MIC = 100 µg/mL), A. niger (MIC = 100 µg/mL), Candida albicans (MIC = 50 µg/mL) and Cryptococcus neoformans (MIC = 25 µg/mL) with inhibition zones ranging from 6 to 8 mm compared to amphotericin B (inhibition zones = 8–12 mm) and nystatin (inhibition zones = 10–14 mm) using agar diffusion assay [ 17 ].
Ethanol extract of S. alata leaves (5-10 mg/mL) inhibited the growth of Microsporum canis, Trichophyton verrucosum and Epidermophyton floccosum with inhibition zones ranging from 12.05 to 20.50 mm compared to clotrimazole (inhibition zones = 21.0-25.5 mm) using screwed diffusion assay. The extract (1.25-10.00 mg/mL) inhibited the growth of Trichophyton mentagrophytes with inhibition zone ranging from 11.0-19.5 mm [ 19 ].
Antidiabetic activity
Ethanol and methanol extract of S. alata leaves (100 mg/kg) respectively administered orally to streptozotocin-induced diabetic Swiss albino mice (ten weeks) reduced the glucose levels as much as 18.74% and 8.35% respectively, compared to initial glucose levels, after 180 min of glucose load compared to glibenclamide control group (reduced by 132.94 %) [ 20 ].
Anti-inflammatory activity
Hot aqueous extract of both heat-treated and sun-dried S. alata leaves (0.01–1.00 mg/mL) showed inhibitory effect (0.05–98.4% inhibition) on the histamine release from concanavalin A-induced rat peritoneal mast cells [ 21 ].
Hot aqueous extract of heat-treated S. alata leaves (0.3–1.00 mg/mL) showed significant (p < 0.05) anti-inflammatory activity on cyclooxygenase-1 (COX-1) with with inhibition rate of 92.5–96.6% compared to sun-dried S. alata leaves (86.6–100.5% inhibition rate). The extract (0.03–1.00 mg/mL) significantly (p < 0.05) showed anti-inflammatory activity on cyclooxygenase-2 (COX-2) with inhibition rate of 15.5–84.6% compared to sun-dried S. alata leaves (6.10–72.9% inhibition rate) [ 21 ].
Hot aqueous extract of heat-treated S. alata leaves (0.01–0.1 mg/mL) showed significant (p < 0.05) inhibition on 5-lipoxygenase with inhibition rate of 26.9–84.1% compared to sun-dried S. alata leaves (3.2–64.9% inhibition rate) [ 21 ].
Anti-oxidant activity
Aqueous extract of S. alata leaves (2.66–7.09 μg/mL) showed anti-oxidant activity with 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity inhibition concentration at 50% of growth (IC50) of 2.25 ±0.28 μg/mL compared to ascorbic acid (IC50 = 3.99 ±0.09 μg/mL) and Trolox (IC50 = 4.50 ±0.08 μg/mL) [ 22 ].
Weight reduction activity
Methanol extract of S. alata (3.0%) with high fat diet feed was administered orally to female ICR mice (three weeks old) for seven weeks. The extract significantly (p < 0.05) reduced the body weight gain (≈ 8.0 g) and parametrial fat weight (≈ 0.7088 g) compared to high fat diet control group (body weight gain ≈ 18.0 g, parametrial fat weight ≈ 1.3187 g) [ 23 ].
Clinical studies
A prospective study of the healing effect of S. alata leaves on Pityriasis versicolor infection was conducted involving 200 individuals (male and female) aged 16–60 years for a period of ten years. The extract applied topically as single application (two hours before sleep) over the infected area showed healing effect at third week of treatment with repigmentation occurring at third month. The lesion required four to ten months to completely heal. Recurrence occurred for a maximum period of one year [ 24 ].
SAFETY INFORMATION
Preclinical studies (Toxicology studies)
Ethanol extract of S. alata leaves (1000, 2000 and 3000 mg/kg) was administered orally as single dose to female Swiss albino mice (22–30 g). After 14 days, no toxicity effect was observed (LD 50 > 3000 mg/kg) [ 20 ].
Methanol extract of S. alata leaves (1000, 2000 and 3000 mg/kg) was administered orally as single dose to female Swiss albino mice (22–30 g). After 14 days, there was no toxicity effect was observed (LD50> 3000 mg/kg) [ 20 ].
Oral single dose acute toxicity study on female Sprague Dawley rats (aged between 8 and 12 weeks old) using aqueous extract of S. alata leaves showed no toxicity effect on the parameters observed, including behaviors, body weight, food, and water intake. All rats were observed for 14 days prior to necropsy. No death was found throughout the study period. Necropsy revealed no significant abnormality. No-observed-adverse effect level(NOAEL) is more than 2,000 mg/kg body weight [ 25 ]
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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