Ara tanah Aerial
Euphorbia hirta L.
Euphorbiaceae
Figure 1 : Euphorbia hirta. (a) Whole Plant; (b) part of a plant showing opposite leaves arrangement; (c) inflorescence; (d) abaxial leaf; (e) adaxial leaf. (Photos courtesy of Centre for Herbal Standardization, USM, 2017)
DEFINITION
Ara tanah aerial consists of the powder of dried leaves, stems and flowers of Euphorbia hirta L. (Euphorbiaceae).
SYNONYM
Chamaesyce gemella (Lag.) Small, Chamaesyce hirta (L.) Millsp., Chamaesyce karwinskyi (Boiss.) Millsp., Chamaesyce rosei Millsp., Desmonema hirta (L.) Raf., Ditritea hirta (L.) Raf., Euphorbia bancana Miq., Euphorbia capitata Lam., Euphorbia chrysochaeta W.Fitzg., Euphorbia gemella Lag., Euphorbia globulifera Kunth, Euphorbia karwinskyi Boiss., Euphorbia nodiflora Steud., Euphorbia obliterata Jacq [ 1 ].
VERNACULAR NAMES
Pill bearing spurge (English); ara tanah (Malay); fei yang cao (Chinese); amampatchaiarisi (Tamil) [ 2 , 3 , 4 , 5 ].
CHARACTER
IDENTIFICATION
Plant Morphology
Euphorbia hirta is an annual, unarmed, erect, pilose herb up to 6–60 cm tall, white latex in all parts, with rather fleshy stems and branches. Stems glabrous or hairy all round. Leaves dark green, 2–4 cm long, opposite, oblong-lanceolate, minutely dentate or serrulate and unequal at the base, acute to obtuse with purple spots, sparsely hairy above, lower surface loosely covered with long hairs. Inflorescence axillary or terminal, composed of peduncled, globose clusters of cyathia; cyathia with appressed-hairy involucres and 4 minute, green or purplish glands bearing a narrow appendage, anthers yellow; capsule acutely 3-lobed, broadly ovoid-pyramidal, about 1 mm x 1.2 mm, appressed-hairy. Flowers are arranged in heads; look like a single flower; each head comprising 4–5 bracts often lobed or glanded surrounding a group of male flower in the centre; male reduced to 1 stamen each, without sepals or petals; females with a 3-chambered ovary with 3 styles often hanging out of heads, pedicel glabrous; ovary and appressed, pubescent. Fruits tiny, 1.5 mm long, yellow, 3-celled capsule, each of carpel contains a single reddish, four-sided, transversely wrinkled seed. Seeds oblong with slightly transversely ribbed when dry. Prime root thick about 5 mm, branch root fibrous [ 2 , 7 ].
Microscopy
Powdered material consists of fragment of parenchyma cells; testa is formed by lignified, thick-walled macrosclereid, in surface view; trichome simple, uniseriate and multicellular; fragments of vascular tissue with spiral vessel, reticulate or bordered-pitted thickenings; starch granules are mostly simple and numerous; isolated fibres; fragment of epidermis cells.
Figure 2 : Microscopic characters of Euphorbia hirta aerial powder of 0.355 mm size. (a) Parenchyma cells (magnification 10x); (b) macrosclereid of testa (magnification 20x); (c) multicellular trichome (arrow) (magnification 20x); (d) spiral vessel (arrow) (magnification 20x); (e) reticulate vessel (arrow) (magnification 20x); (f) pitted vessel (magnification 20x); (g) starch granules (arrow) (magnification 20x); (h) fibre (arrow) (magnification 10x); (i) epidermis cells (magnification 10x). [Scale bars: a = 10 µm; b, c, d, e, f, g = 20 µm; h, i = 50 µm]
Thin Layer Chromatography (TLC)
Figure 3 :TLC chromatogram of quercitrin (S), methanol extract of Euphorbia hirta dried aerial powder (L) observed under 366 nm after derivatization.
| Test Solutions | Weigh about 1.0 g of E. hirta dried aerial powder of 0.355 mm size in 100 mL flask. Add 10 mL of methanol and reflux the mixture for 45 min. Filter the mixture and use the filtrate as test solution. |
| Standard solution | Dissolve quercitrin standard [CAS no.: 522-12-3] in methanol to produce a standard solution of 1.0 mg/mL. |
| Stationary Phase | HPTLC Silica gel 60 F254, 10 x 10 cm |
| Mobile phase | Ethyl acetate : water : formic acid : acetic acid (40 : 8 : 2 : 5.5) (v/v/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 | Weigh about 1.0 g of E. hirta dried aerial powder of 0.355 mm size in 100 mL flask. Add 10 mL of methanol and reflux the mixture for 45 min. Filter the mixture and use the filtrate as test solution. | |||||||||||||||||||||
| Standard solution | Dissolve quercitrin standard [CAS no.: 522-12-3] in methanol to produce a standard concentration of 0.2 mg/mL. | |||||||||||||||||||||
| Chromatographic system |
Detector: UV 330 nm Column: C18 (150 X 4.6 mm, 3.5 µm) (preferably Zorbax SB-C18) Column oven temperature: 25oC Flow rate: 1 mL/min Injection volume: 10 µL |
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| Mobile Phase (gradient mode) |
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| System suitability requirement |
Perform at least five replicate injections of the quercitrin standard solution (0.2 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 : Whole HPLC chromatogram of (a) quercitrin standard solution (0.2 mg/mL) at tr = 10.5 min and (b) methanol extract of Euphorbia hirta dried aerial powder showing peak corresponding to quercitrin standard solution at tr = 10.5 min.
(a)
(b)
Figure 5 : HPLC chromatogram highlighting the elution region of quercitrin in (a) quercitrin standard solution (0.2 mg/mL) and (b) methanol extract of Euphorbia hirta dried aerial powder showing peak corresponding to quercitrin standard solution at tr = 10.5 min.
Figure 6 : UV spectrum of quercitrin standard solution (0.2 mg/mL) and methanol extract of Euphorbia hirta dried aerial powder.
PURITY TESTS
The purity tests except foreign matter test are based on E. hirta dried aerial powder of 0.355 mm particle size.
| Foreign Matter |
| Not more than 2% |
| Ash Contents | |
| Total ash | Not more than 11% |
| Acid-insoluble ash | Not more than 1% |
| Water-soluble ash | Not less than 4% |
| Loss on Drying |
| Not more than 10% |
| Extractive Values | |
| Water-soluble extracts | |
| Hot method | Not less than 21% |
| Cold method | Not less than 15% |
| Ethanol-soluble extracts | |
| Hot method | Not less than 6% |
| Cold method | Not less than 3% |
SAFETY TESTS
The safety tests are based on E. hirta dried aerial 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
Methanol extract of E. hirta leaves was found to contain diterpene (phytol) and flavonoid (myricitrin) [ 8 , 9 ].
Methanol extract of E. hirta aerial part was found to contain flavonol glycosides (afzelin, quercitrin, myricitrin), diterpenoids (phytol, tocopherol), terpenes (megastigmatrienone, neophytadiene), triterpene (squalene), triterpenoids (cycloartenol, campesterol, clionasterol, alnulin, beta-amyrin, alpha-amyrin), fatty acids (palmitic acid, linoleic acid, methyl linolenate, 2-monopalmitin, 2-monostearin, methyl palmitate), flavonoid (epicatechin 3-gallate) and phenolic (caffeic acid) [ 10 , 11 , 12 ].
Ethanol (85%) extract of E. hirta aerial part was found to contain phenols (scopoletin, scoparone, isoscopoletin, gallic acid) and flavonoids (quercetin, isorhamnetin, pinocembrin, kaempferol, luteolin) [13].
MEDICINAL USES
Uses described in folk medicine, not supported by experimental or clinical data
Traditionally, the latex of E. hirta is used to alleviate eye problems. The juice is given three days after child-birth for meroyan. Poultice of E. hirta is applied to the sores on the legs, bruises and wounds [ 3 ].
Biological and pharmacological activities supported by experimental data
Antioxidant activity
Ethyl acetate fraction from the methanol extract of E. hirta dried leaves (10 – 100 µg/mL) showed hydroxyl radical scavenging activity with 50% inhibition concentration (IC50) valueof 33.25 µg/mL respectively compared to butylated hydroxytoluene (BHT) with (IC50 = 43.87 µg/mL) using hydroxyl radical scavenging assay [ 9 ].
Ethyl acetate fraction from the methanol extract of E. hirta dried leaves (10 – 100 µg/mL) showed 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity with IC50 valueof 13.93 µg/mL compared to BHT with (IC50 = 20.64 µg/mL) using DPPH assay [ 9 ].
Methanol extract of E. hirta dried leaves powder (0.031 – 2.0 mg/mL) showed maximum radical scavenging activity at 1 mg/mL (72.96 ± 0.78%) with (IC50) valueof 0.803 mg/mL comparable to (BHT) with (75.13 ± 0.75%) and IC50 value of 0.794 mg/mL using DPPH assay [ 14 ].
Ethanol extract of E. hirta dried aerial powder (31.25 – 500 µg/mL) showed reducing power activity with 50% effective concentration (EC50) valueof 6.18 ± 2.52 µg/mL compared to ascorbic acid with (EC50 = 36.1 ± 0.49 µg/mL) using reducing power assay [ 15 ].
Ethanol, ethyl acetate and dichloromethane extract of E. hirta dried aerial powder showed β-carotene bleaching activity with inhibition percentage of 94.32%, 93.75% and 96.15% respectively, comparable to BHT (93.2%) using β-carotene linoleic acid bleaching assay [ 15 ].
Antimicrobial activity
Methanol extract of E. hirta dried leaves and flowers (100 mg/mL) showed antibacterial activity against Gram-positive bacteria Staphylococcus aureus with inhibition zone (IZ) of 28 mm diameter comparable to chloramphenicol (30 µg/mL) (IZ = 23 mm) using disc diffusion method [ 16 ].
Methanol extract of E. hirta dried leaves and flowers (100 mg/mL) showed antibacterial activity against Gram-positive bacteria Micrococcus sp. with inhibition zone (IZ) of 29 mm diameter comparable to chloramphenicol (30 µg/mL) (IZ = 26 mm) using disc diffusion method [ 16 ].
Ethanol extract of E. hirta dried aerial part (25 mg/mL) showed antibacterial activity against Gram-negative bacteria Escherichia coli with inhibition zone (IZ) of 21 mm diameter comparable to ampicillin (40 µg/mL) (IZ = 17 mm) using agar well diffusion method [ 17 ].
Ethanol extract of E. hirta dried aerial part (25 mg/mL) showed antibacterial activity against Gram-negative bacteria Pseudomonas aeruginosa with inhibition zone (IZ) of 23 mm diameter comparable to ampicillin (40 µg/mL) (IZ = 16 mm) using agar well diffusion method [ 17 ].
Methanol extract of E. hirta dried leaves (100 mg/mL) showed antifungal activity against Candida albicans with inhibition zone (IZ) of 21 mm diameter comparable to miconazole nitrate (30 µg/mL) (IZ = 21 mm) using disc diffusion method. The minimum fungicidal concentration of the methanol extract is 3.13 mg/mL [ 16 ].
Antidiabetic activity
Ethanol extracts of E. hirta dried leaves (250 and 500 mg/kg) were administered orally to streptozotocin-induced diabetic albino mice of either gender (30 – 35 g) for duration of 15 days. The extracts showed significant (p < 0.001) decrease in blood glucose level at day-15 (80.6 and 77.6 mg/dL, respectively) compared to diabetic control group (192.5 mg/dL) and glibenclamide treated group (75.73 mg/dL) [ 18 ].
Ethanol extract of E. hirta dried stems (500 mg/kg) was administered orally to streptozotocin-induced diabetic albino mice of either gender (30 – 35 g) for duration of 15 days. The extract showed significant (p < 0.001) decrease in blood glucose level at day-15 (80.3 mg/dL) compared to diabetic control group (192.5 mg/dL) and glibenclamide treated group (75.73 mg/dL) [ 18 ].
Anti-anaphylactic activity
Ethanol (95%) extract of E. hirta dried aerial part (500 mg/kg) was administered orally to anaphylaxis-induced Wistar rats (150+5 g) one hour before passive cutaneous anaphylaxis (PCA) challenge. The extracts inhibited the PCA reaction in dose dependent manner (75% inhibition) compared to cetirizine (10 mg/kg) with 93% inhibitory effect [ 19 ].
Ethanol (95%) extracts of E. hirta dried aerial part (500 mg/kg) administered orally to ovalbumin-induced BALB/c mice (25+5 g) for 10 days. At three hours, the extracts inhibited the paw edema in dose dependent manner (70% inhibition) and maintained at the same level after 24h compared to cetirizine (3h: 76%; 24h: 60%) [ 19 ].
Ethanol (95%) extract of E. hirta dried aerial part (100 – 125 µg) significantly (p < 0.001) inhibit histamine release induced by the compound 48/80 from rat peritoneal mast cells (RPMC) with inhibition percentage of 100% – 120% compared to ketotifen (3.75 µg; 85%) [ 20 ].
Clinical studies
Information and data have not been established.
SAFETY INFORMATION
Preclinical studies (Toxicology studies)
14-day oral toxicity study
Ethanol extracts of E. hirta dried leaves, flowers and stem (2000 mg/kg) administered orally to male and female albino mice (30 – 35 g) showed no mortality after 14 days with lethal dose at 50%, LD50 > 2000 mg/kg [ 18 ].
Oral single dose acute toxicity study on female Sprague Dawley rats (aged between 7 and 12 weeks old) using aqueous extract of E. hirta aerial showed no toxic effect on the parameters observed, including behavior, 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 but several histological lesions were found. Approximate lethal dose (LD50) is not more than 2,000 mg/kg body weight [21].
Others (Adverse reaction, contraindication, side effect, warning, precaution)
Information and data have not been established.
DOSAGE
In Chinese Pharmacopoeia, the recommended dosage is 6-9 g, appropriate amount for topical application and decoction for bathing [ 4 ].
STORAGE
Store below 30°C. Protect from light and moisture.
REFERENCES
- The Plant List. [Internet] Euphorbia hirta L.; 2013 [cited on 17th January 2017]. Available from: http://www.theplantlist.org/tpl1.1/record/kew-80144.
- PROSEA (Plant Resources of South-East Asia) Foundation Bogor, Indonesia. [Internet] Euphorbia hirta L; 1999 [cited on 17th January 2017]. Available from: http://proseanet.org/prosea/e-prosea_detail.php?frt=&id=254.
- Burkill IH. A dictionary of the economic product of the Malay Peninsula. Vol. 1. London: Published on behalf of the Governments of the Straits Settlements and Federated Malay States by the Crown Agents for the Colonies. 1935; p.334-339.
- Chinese Pharmacopoeia Commission. Pharmacopoeia of the People’s Republic of China. Vol. 1. China Medical Science Press. 2010; p.180.
- Quattrocchi UFLS. CRC world dictionary of medicinal and poisonous plants: common names, scientific names, eponyms, synonyms, and etymology. Vol. III E-L. United States: CRC Press. 2012; p.174.
- N’Guessan BYF, Alida EO, Martin K, Rokia S, Dieneba KB. Pharmacognostic study of Euphorbia hirta Linn.: An hyperglycaemic plant. International Journal of Sciences and Research. 2017;6(1):410-416.
- ASEAN Committee. Standard of ASEAN herbal medicine. Vol. 2. Jakarta: ASEAN Countries. 2004; p.69-78.
- Chitra M, Muga V, Sasikumar D, Awdah MA. Screening of phytochemical and in vitro activity of Euphorbia hirta L. Journal of Chemical and Pharmaceutical Research. 2011;3(6):110-114.
- Ashish K, Ansar P, Snehal D, Dheeraj B. Free radical scavenging activity of Euphorbia hirta Linn. Leaves and isolation of active flavonoid myricitrin. Asian Journal of Pharmaceutical and Clinical Research. 2010;3(3):234-237.
- Liu Y, Murakami N, Ji H, Pedro A, Zhang S. Antimalarial flavonol glycosides from Euphorbia hirta. Pharmaceutical Biology. 2007;45(4):278-281.
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- Shanmugapriya P, Roziahanim M. Anti-infective potential of caffeic acid and epicatechin 3-gallate isolated from methanol extract of Euphorbia hirta (L.) against Pseudomonas aeruginosa. Natural Product Research. 2015;29(18):1766-1769.
- Wu Y, Qu W, Geng D, Liang JY, Luo YL. Phenols and flavonoids from aerial part of Euphorbia hirta. Chinese Journal of Natural Medicines. 2012;10(1):40-42.
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- Shanmugapriya P, Roziahanim M, Suthagar PP, Wei Cai L, Surash R. Antiradical and cytotoxic activities of varying polarity extracts of the aerial part of Euphorbia hirta L. Journal of Chemistry. 2013:6.
- Mohammad ABR, Zakaria Z, Sreenivasan S, Lachimanan YL, Santhanam A. Assessment of Euphorbia hirta L. leaf, flower, stem and root extracts for their antibacterial and antifungal activity and brine shrimp lethality. Molecules. 2010;15:6008-6018.
- Sudhakar M, Rao Ch.V, Raju DB, Venkateswarlu Y. Antimicrobial activity of Caesalpinia pulcherrima, Euphorbia hirta and Asystasia gangeticum. Fitoterapia. 2006;77:378-380.
- Sunil K, Rashmi, Kumar D. Evaluation of antidiabetic activity of Euphorbia hirta Linn. in streptozotocin induced diabetic mice. Indian Journal of Natural Products and Resources. 2010;1(2):200-203.
- Youssouf MS, Kaiser P, Tahir M, Singh GD, Singh S, Sharma VK, Satti NK, Haque SE, Johri RK. Anti-anaphylactic effect of Euphorbia hirta. Fitoterapia. 2007;78:535-539.
- Singh GD, Kaiser P, Youssouf MS, Singh S, Khajuria A, Koul A, Bani S, Kapahi BK, Satti NK, Suri KA, Johri RK. Inhibition of early and late phase allergic reactions by Euphorbia hirta L. Phytotherapy Research. 2006;20:31.
- Elda Nurafnie IR, Nor Azlina Z, Umi Rubiah SZ, Janice CSW, Tavinraj R, Nurmaziah MS, Teh BP. Acute oral toxicity study of selected Malaysian medicinal herbs ( Euphorbia hirta aerial) on Sprague Dawley rats. Institute for Medical research, Ministry of Health; 2020. Report no.: NON-GLP/2020/01/01.
