Malaysian Herbal Monograph

Kucing galak aerial

Acalypha indica L.

Euphorbiaceae

Figure 1 : Acalypha indica. (a)-(b) Whole plant; (c) leaves arrangement; (d) abaxial and adaxial leaf surface; (e) inflorescence. (Photos courtesy of Centre for Herbal Standardization, 2018)

DEFINITION

Kucing galak aerial consist of the powder of dried leaves, stems and flowers of Acalypha indica L.

SYNONYM

Acalypha bailloniana Mull.Arg., Acalypha chinensis Benth., Acalypha cupamenii Dragend., Acalypha decidua Forssk., Acalypha fimbriata Baill., Acalypha somalensis Pax, Acalypha somalium Müll.Arg., Acalypha spicata Forssk., Cupamenis indica (L.) Raf., Ricinocarpus baillonianus (Müll.Arg.) Kuntze, Ricinocarpus deciduus (Forssk.) Kuntze, Ricinocarpus indicus (L.) Kuntze [ 1 ].

VERNACULAR NAMES

Indian acalypha, India nettle, three-seeded mercury (English); chika mas, kucing galak, lis-lis, rumput lislis, tjeka mas (Malay); tie xian cai (Chinese); kupaaimeni (Tamil) [ 2 , 3 , 4 ].

CHARACTER

ColourGreenish brown
OdourCharacteristic and admissible odor [ 5 ]
TasteSlightly bitter [ 4 ]

IDENTIFICATION

Plant Morphology

Acalypa indica is an erect and annual herb, 0.3–1.5 m in height. Stems are brownish when mature and younger parts are green, sparsely hairy, terete, 2–10 mm in thickness. Leaves are simple and alternate, dull to dark green to brownish; brittle when dry; petiole 1–7 cm, lower leaves with longer petiole, pubescent; lamina 2–5 cm long and 1–4 cm broad, ovate to rhombic ovate, tip acute, base cuneate, pale green below and dark green above, margin serrate and hairy; veins 5–7 pairs, generally alternate, usually 3 veins arising from the base, prominent and hairy below; midrib slightly raised on the upper surface, and prominent on the lower surface. Inflorescence are axillary, stalked, spike, 1–7 cm long. Flowers unisexual, green, subsessile and encircled by a leafy, orbicular serrate bract of about 4 mm long and 5–8 mm broad. Female flowers 5 to 15, basal, 2 mm across. Male flowers numerous, minute; spike usually terminating in an allomorphic flower. Fruits capsules, small and green. Seeds minute, ovoid and pale brown. Roots are vertical and branched; 2–8 mm in thickness, tortuous, rough; colour varies from grey to brown when dry, broken surface creamy yellow; fracture giving rise to a cloud of dusty particles; no characteristic smell; bitter [ 4 , 5 ].

Microscopy

Powdered material consists of fragments of pitted, annular, spiral and scalariform vessels; covering trichomes consist of whole unicellular and multicellular with thin and smooth walls; calcium oxalate consist of numerous type of solitary and druses crystals; fragment of thin-walled parenchyma cells; fragment of idioblast tannin cells; fragments of lower epidermis in surface view with wavy to sinuous anticlinal walls and abundant paracytic type of stomata; fragment of lignified fibre [ 4 , 5 ].

Figure 2 : Microscopic characters of Acalypha indica aerial powder of 0.106 mm size. (a) Pitted vessel (magnification 40x); (b) spiral vessel (magnification 40x); (c) spiral vessel (magnification 40x); (d) simple unicellular trichome (magnification 20x); (e) simple multicellular trichome (magnification 40x); (f) solitary crystal (arrow) (magnification 40x); (g) druses crystal (magnification 40x); (h) idioblast cell (arrow) (magnification 40x); (i) idioblast tannin cell (magnification 40x); (j) paracytic stomata (arrow) (magnification 40x); (k) lignified parenchyma (magnification 40x). [Scale bars: a = 200 µm; b, c, f, g, h, I, k = 20 µm; d, e, j, l = 50 µm]

Chemical Tests 

Observation of solution after treatment with various reagents:

Test for the presence of steroidsBluish green
Test for the presence of phenolicGreen

Thin Layer Chromatography (TLC)

Figure 3 : TLC chromatogram of biorobin (S), methanol extract of Acalypha indica dried aerial parts (L) observed under (a) UV at 366 nm after derivatization

Test Solutions Weigh about 1.0 g of A. indica  dried aerial powder of 0.335 mm size in 50 mL of conical flask. Add 10 mL of methanol. Sonicate the mixture for 15 min. The mixture was then centrifuged and supernatant was collected.
Standard solution Dissolve biorobin standard [CAS no. 1729-56-2] in methanol to produce concentration of 0.5 mg/mL. [Vortex standard for 15 sec].
Stationary Phase HPTLC Silica gel 60 F254 10 x 10 cm
Mobile phase Ethyl acetate : water : formic acid : acetic acid; (34 : 7 : 2.5 : 5.5) (v/v/v/v)
Application
  1. Biorobin standard solution (S); 1 µL, as a band
  2. Methanol extract of A. indica dried aerial powder (L); 10 µL, as a band
Development distance 8 cm
Drying Air drying
Detection
  1. UV at 366 nm after derivatisation with NP-PEG reagent.Derivatizing reagent preparation: Derivatized with natural product reagent and polyethylene glycol 4000 reagent. Heat for 5 min at 100°C.

    Natural Product Reagent (NP)

    Dissolve 1.0 g of 2-aminoethyldiphenylborinate in 100 mL of methanol.

    Polyethylene glycol 4000 (PEG)

    Dissolve 5.0 g of polyethylene glycol 4000 in 100 mL ethanol.

High Performance Liquid Chromatography (HPLC)

Test solution Weigh about 1.0 g of A. indica dried aerial powder of 0.335 mm size in 50 mL of conical flask. Add 10 mL of methanol. Sonicate the mixture for 15 min. The mixture was then centrifuged and supernatant was collected.
Standard solution Dissolve biorobin standard [CAS no. 1729-56-2] in methanol to produce a standard concentration of 0.05 mg/mL. Vortex standard for 15 sec.
Chromatographic system

Detector: 350 nmColumn: C18 (150 X 4.6 mm, 3.5 µm) (Zorbax SB-C18 if necessary)

Column oven temperature: 25oC

Flow rate: 1 mL/min

Injection volume:20 µL

Mobile Phase (gradient mode)

Run Time

(min)

A – 0.1% formic acid in water
(%)

B – 0.1% formic acid in methanol
(%)

0

65

35

25

65

35

29

0

100

34

0

100

34.1

65

35

40

65

35

System suitability requirement

Perform at least five replicate injections of the standard solutions (0.05 mg/mL). The requirements of the system suitability parameters are as follow:

  1. Symmetry factor (As) for biorobin standard is not more than 1.5.
  2. Percentage of relative standard deviation (RSD) of the retention time (tr) for biorobin standard is not more than 2.0%.
Acceptance criteria
  1. Retention time (tr) of biorobin standard in the test solution is similar to the tr of the standard solution.
  2. The ultraviolet (UV) spectrum of biorobin standard in the test solution is similar to the UV spectrum of biorobin standard in the standard solution (optional supportive data).
4a

(a)

4b

(b)

Figure 4 : Whole HPLC chromatogram of (a) biorobin standard standard solution (0.05 mg/mL) at tr= 25.3 min and (b) methanol extract of Acalypha indica dried aerial powder showing peak corresponding to biorobin standard standard solution at tr= 25.2 min.

5a

(a)

5b

(b)

Figure 5 : HPLC chromatogram highlighting the elution region of (a) biorobin standard standard solution (0.05 mg/mL) at tr= 25.3 min and (b) methanol extract of Acalypha indica dried aerial powder showing peak corresponding to biorobin standard standard solution at tr= 25.2 min.

6

Figure 6 : UV spectrum of biorobin standard solution (0.05 mg/mL) and methanol extract of Acalypha indica dried aerial powder.

PURITY TESTS

The purity tests, except foreign matter test are based on A. indica dried plant part powder of 0.355 mm particle size.

Foreign Matter
Not more than 2%
Ash Contents
Total ash Not more than 15%
Acid-insoluble ash Not more than 1%
Water soluble ash Not less than 4 %
Loss on Drying
Not more than 8%
Extractive Values
Water-soluble extracts
Hot method Not less than 16%
Cold method Not less than 12%
Ethanol-soluble extracts
Hot method Not less than 5%
Cold method Not less than 2%

SAFETY TESTS

The safety tests are based on A. indica dried plant part 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 A. indica dried flowers and leaves were found to contain kaempferol glycosides (mauritianin, clitorin, nicotiflorin and biorobin) [ 6 ].

Methanol extract of A. indica dried aerial parts were found to contain pyridone glucosides (acalyphin, epiacalyphin, noracalyphin, epinoracalyphin, acalyphin amide, epiacalyphin amide cycloside, ar-acalyphidone and seco-acalyphin) [ 7 ].

Methanol extract of A. indica dried leaves were found to contain fatty acids (pentadecanoicacid, 14-methyl-, methyl ester, n-hexadecanoic acid, 10-octadecanoic acid, methyl ester, heptadecanoic acid, 16-methyl-methyl ester, hexadecanoic acid, 14-methyl-methyl ester, nonadecanoic acid, 18-oxo, methyl ester, 14-methylhexadecanoic acid, picolinyl ester, 9-octadecenoic acid [Z], 2-hydroxy-1-(hydroxymethyl) ethyl ester) and non fatty acids (13-hexyloxacyclotridec 10-en-2-one, imidazole, 4-fluoro-5-hydroxyazomethyl-, 2-propenoic acid, 3-[5-acetyl-2,2-dimethlcyclopenthyl],methyl ester, [1a(E),5a]-, 1-oxaspiro [2,5] ocatane, 5,5-dimethyl-4-(3-methyl-1,3-butadienyl)-, 2-formyl-5,7,dimethyl-1,2,3,4-tetrahydropyrimido(3,4-a) indole) [ 8 ].

Essential oil of A. indica dried leaves were found to contain alcohol ((Z)-3-hexen-1-ol, (E)-3,7-dimethyl-2,6-octadien-1-ol)) alkanes (eicosane, heneicosane, docosane) , fatty aldehydes (tridecanal, tetradecanal, palmitaldehyde) fatty acids (9,12,15-octadecatrienoic acid) terpene (10-(acetylmethyl)-(+)-3-carene, phytol isomer) sesquiterpenoids (neophytadiene) [ 9 ].

MEDICINAL USES

Uses described in folk medicine, not supported by experimental or clinical data

Traditionally used as a laxative, for which purpose the plant is boiled and the extract is consumed [ 10 ].

Biological and pharmacological activities supported by experimental data

Antioxidant activity

Chloroform extract of A. indica dried aerial exhibited 2,2’-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity with 50% inhibition concentration (IC50) value of 6.31 mg/mL compared to trolox (IC50 = 1.32 mg/mL) using ABTS assay [ 11 ].

Hexane extract of A. indica dried aerial exhibited ABTS radical scavenging activity with 50% inhibition concentration (IC50) value of 6.13 mg/mL compared to trolox (IC50 = 1.32 mg/mL) using ABTS assay [ 11 ].

Methanol extract of A. indica dried aerial exhibited ABTS radical scavenging activity with 50% inhibition concentration (IC50) value of 6.37 mg/mL compared to trolox (IC50 = 1.32 mg/mL) using ABTS assay [ 11 ].

Antimicrobial activity

Methanol extract of A. indica dried leaves (0.2 mg/mL) showed antifungal activity against Candida albicans with inhibition zone (IZ) of 25 mm diameter comparable to clotrimazole (0.01 mg/mL) (IZ = 20 mm) using disc diffusion method [ 12 ].

Methanol extract of A. indica dried stem (0.2 mg/mL) showed antifungal activity against C. albicans with inhibition zone (IZ) of 20 mm diameter comparable to clotrimazole (0.01 mg/mL) (IZ = 20 mm) using disc diffusion method [ 12 ].

Chloroform extract of A. indica dried leaves (0.2 mg/mL) showed antifungal activity against Aspergillus niger with inhibition zone (IZ) of 22 mm diameter comparable to clotrimazole (0.01 mg/mL) (IZ = 22 mm) using disc diffusion method [ 12 ].

Methanol extract of A. indica dried stem (0.2 mg/mL) showed antifungal activity against A. niger with inhibition zone (IZ) of 18 mm diameter comparable to clotrimazole (0.01 mg/mL) (IZ = 22 mm) using disc diffusion method [ 12 ].

Hexane extract of A. indica dried leaves (0.2 mg/mL) showed antibacterial activity against Escherichia coli with inhibition zone (IZ) of 25 mm diameter comparable to clotrimazole (0.01 mg/mL) (IZ = 22 mm) using disc diffusion method [ 12 ].

Chloroform extract of A. indica dried stem (0.2 mg/mL) showed antibacterial activity against E. coli with inhibition zone (IZ) of 22 mm diameter comparable to clotrimazole (0.01 mg/mL) (IZ = 22 mm) using disc diffusion method [ 12 ].

Methanol extract of A. indica dried leaves (5 mg/disc) showed antibacterial activity against Staphylococcus aureus with inhibition zone (IZ) of 15.8 mm diameter comparable to streptomycin (10 µg/disc) (IZ = 14 mm) using disc diffusion method [ 13 ].

Methanol extract of A. indica dried leaves (5 mg/disc) showed antibacterial activity against Bacillus cereus with inhibition zone (IZ) of 14 mm diameter comparable to streptomycin (10 µg/disc) (IZ = 12 mm) using disc diffusion method [ 13 ].

Methanol extract of A. indica dried leaves (5 mg/disc) showed antibacterial activity against Strepcoccus faecalis with inhibition zone (IZ) of 16.5 mm diameter comparable to streptomycin (10 µg/disc) (IZ = 13 mm) using disc diffusion method [ 13 ].

Anthelmintic activity

Ethanol extract of A. indica dried leaves extract (25, 50 and 100 mg/mL) showed significant (p < 0.05) anthelmintic activity in dose dependent manner towards Pheretima posthuma (use as model for anthelmintic screening activity due to its anatomical and physiological resemblance with intestinal roundworm parasites of human being). The ethanol extract (100 mg/mL) demonstrated paralysis time (P: 10 min) and death time (D: 29 min) of worms compared to positive control, piperazine citrate (P: 8 min and D: 20 min) [ 14 ].

Cytotoxicity activity

Methanol extract of A. indica dried aerial parts showed cytotoxic activity on NCI-H187-small lung cancer cell lines with IC50 values of 25 µg/mL compared to ellipticine (IC50 = 0.88 µg/mL) and doxorubicin (IC50 = 0.88 µg/mL) using Resazurin Microplate Assay (REMA) [ 11 ].

Hepatoprotective activity

Methanol extract of A. indica dried aerial parts (300 mg/kg) was administered orally to Wistar albino rats of either sex (175 – 225 g) in a single daily dose for three days.  Thioacetamide as hepatoxicity inducer was administered orally on the second day 30 min after the administration of the extract. After 48 hr of thioacetamide administration, the extract showed hepatoprotective activity by decreasing the glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), alkaline phosphatase (ALKP) (GOT: 140.17 ± 7.67 IU/L; GPT: 84.31 ± 28.70 IU/L; ALKP: 427.48 ± 73.38 IU/L) compared to silymarin (GOT: 152.67 ± 7.61 IU/L; GPT: 77.71 ± 27.56 IU/L; ALKP: 385.92 ± 91.27 IU/L) [ 15 ].

Clinical studies

Information and data have not been established.

SAFETY INFORMATION

Preclinical studies (Toxicology studies)

14-days toxicity study

Methanol extract of A. indica dried aerial parts (300 and 2000 mg/kg) administered orally to Wistar albino rats of either sex (175 – 225 g) for 14 days showed no mortality with lethal dose at 50% (LD50) > 2000 mg/kg [ 15 ].

Oral single dose acute toxicity on female Sprague Dawley rats (aged between 7 and 12 weeks old) using aqueous extract of A. Indica 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 . No-observed-adverse -effect level (NOAEL) is not more than 2,000 mg/kg body weight [ 16 ]

Others (Adverse reactions, contraindications, side effects, warning, precautions)

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

  1. The plant List. [Internet] Acalypha indica L.; [cited on 1st March 2018]. Available from: http://www.theplantlist.org/tpl1.1/record/kew-713?ref=tpl1
  2. 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.36-37.
  3. Goh Kong Ling. Malaysian Herbs Chinese Edition. 2000;p.186.
  4. Government of India Ministry of Health & Family Welfare Department of Ayush New Delhi. The Ayurvedic Pharmacopoeia of India. Part I, Volume VI, First Edition. India: Department of Ayush New Delhi. 20118:63-65.
  5. Takle VV, Savad RV, Kandalkar AM, Akarte AM, Patel AM. Pharmacognostic and phytochemical investigations of aerial parts of Acalypha indica Linn. Pharmacognosy Journal. 2011;3(21):33-35.
  6. Nahrstedt A, Hungeling M, Petereit F. Flavonoids from Acalypha indica. Fitoterapia. 2006; 77(6): 484-486.
  7. Hungeling M, Lechtenberg M, Fronczek FR, Nahrstedt A. Cyanogenic and non-cyanogenic pyridone glucosides from Acalypha indica (Euphorbiaceae). Phytochemistry. 2009; 70: 270-277.
  8. Ravi S, Shanmugam B, Subbaiah GV, Prasad SH, Reddy KS. Identification of food preservative, stress relief compounds by GC-MS and HR-LC/1-TOF/MS; evaluation of antioxidant activity of A. indica leaves methanolic extract (in vitro) and polyphenolic fraction (in vivo). Journal Food Science Technology. 2017;54(6):1585-1596.
  9. Rahman MS, Hossain R, Saikot FK, Rahman SM, Saha SK, Hong J, Kim KH. Insights into the in vitro germicidal activities of Acalypha indica. Analytical Science & Technology. 2017;30(1):26-31.
  10. 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. 166-167
  11. Sanseera D, Niwatananun W, Liawruangrath B, Liawruangrath S, Baramee A. Antioxidant and anticancer activities from aerial parts of Acalypha indica L. Chiang Mai University Journal of Natural Sciences. 2012; 11(2): 157-168.
  12. Solomon RDJ, Kallidas S, Vimalan J. Isolation, identification and study of antimicrobial property of a bioactive compound in an Indian medicinal plant Acalypha indica (Indian-nettle). World Journal of Microbiology and Biotechnology. 2015; 21: 1231-1236.
  13. Govindarajan M, Jabanesan A, Reetha D, Amsath R, Pushpanathan T, Samidurai K. Antibacterial activity of Acalypha indica L. European Review for Medical and Pharmacological Sciences. 2008; 12:299-302.
  14. Ranju G, Niranjan S, Kumar PS, Kumar PV, Kumar PS. In vitro anthelmintic activity of Acalypha indica leaves extracts. International Journal of Research in Ayuverda & Pharmacy. 2011; 2(1):247-249.
  15. Kumar SVS, Kumar CV, Vardhan AV. Hepatoprotective activity of Acalypha indica Linn against thioacetamide induced toxicity. International Journal of Pharmacy and Pharmaceutical Sciencec. 2013;5(4):356-359.
  16. 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 (Acalypha indica aerial) on Sprague Dawley rats. Institute for Medical Research, Ministry of Health; 2020. Report no.: NON-GLP/2020/03/01.