Malaysian Herbal Monograph

Mahkota dewa Leaves

Phaleria macrocarpa (Scheff.) Boerl.

Thymelaeaceae

Figure 1 : Phaleria macrocarpa. (a) Young tree; (b) cross section of fruits; (c) whole fruit; (d) leaves; (e) adaxial leaf; (f) abaxial leaf. (Photo courtesy of Centre for Herbal Standardization, 2016)

DEFINITION

Mahkota dewa leaves consist of the powder of dried leaves of Phaleria macrocarpa (Scheff.) Boerl.(Thymelaeaceae).

SYNONYM

Phaleria calantha Gilg, Phaleria papuana Warb. ex. K.Schum & Lauterb., Phaleria papuana var. wichmannii (Valeton)Backer, Phaleria wichmannii Valeton [ 1 ].

VERNACULAR NAMES

God’s crown (English); Mahkota dewa (Malay); Huang kuang (Chinese) [ 2 , 3 ].

CHARACTER

ColourYellowish-green
OdourOdourless
TasteBitter

IDENTIFICATION

Plant Morphology

P. macrocarpa is a shrub or an evergreen tree, height ranges from 1–18 m, with 1 m long straight root exuding sap, erect trunk, cylindrical, up to about 15 cm of diameter, smooth or slightly wrinkled brownish bark and dense crown. Leaves simple, oppositely arrangement, elliptic to oblong lanceolate with pointed apex, 7–14 cm long and 3–5 cm broad, coriaceous, dark green in colour; petiole 0.5 cm long. Inflorescence umbel type, peduncle 0.3–2 cm long, borne in the axil of the leaves, carrying 2–5 white sessile flowers. Flowers tubular shaped, 1.5–3.5 cm long, whitish, fragrant, perfect flower with four sepals, four or five petals, 8 stamens and a carpel. Fruits simple, fleshy drupe with one or two seeds, round or elliptical in shape, matured fruit is 3–5 cm in diameter, colour of matured fruit is green and turns to brightly red or maroon when it ripens. Seeds dicotyledonous with exalbuminous seeds type, brown in colour, ovoid, anatropous and exist as 1-2 seeds per fruit [ 4 , 5 ].

Microscopy

The powdered material consists of adaxial epidermal cells with straight to wavy anticlinal wall; fragments of abaxial epidermis with tetracytic stomata; covering trichomes are simple and multicellular with pointed ends; fragments of vascular tissues; druse and solitary crystals are abundant, sometimes found isolated.

Figure 2 : Microscopic characters of Phaleria macrocarpa leaves powder of 0.355 mm size. (a) Adaxial epidermis cell (magnification 40x); (b) abaxial epidermis cell (magnification 40x); (c) multicellular trichome (magnification 40x); (d) druse calcium oxalate crystals (magnification 40x); (e) solitary prism calcium oxalate crystals (magnification 40x). [Scale bars: a-f = 10 µm]

Colour Tests

Observed colour of solution after treatment with various reagents:

HCl (conc.)Green
NaOH 5% (w/v)Green

Thin Layer Chromatography (TLC)

Figure 3 : TLC chromatogram of mangiferin (S), ethanol extract of Phaleria macrocarpa dried leaves powder (L) observed under (a) 366 nm after derivatisation.

Test Solutions

Weigh about 2.0 g of P. macrocarpa dried leaves powder of 0.355 mm size in a 100 mL conical flask. Add 50 mL of hexane and shake the mixture for 4 hr at room temperature. Discard the hexane and reconstitute the raw material with 10 mL of ethanol. Sonicate the mixture for 15 min at room temperature. Centrifuge the mixture for 5 min at 13,000 rpm and use the supernatant as test solution.

Standard solution

Dissolve mangiferin standard [CAS no.:4773-96-0] in ethanol to produce a standard concentration 1.0 mg/mL.

Stationary Phase HPTLC Silica gel 60 F254 10×10 cm
Mobile phase

Chloroform : acetone : formic acid : methanol; (35 : 40 : 8 : 15) (v/v/v/v)

Application
  1. Mangiferin standard solution (S); 1 μL, as a band

  2. Ethanol extract of P. macrocarpa dried leaves powder (L); 2 µL, as a band.

Development distance 8 cm, automatic developing chamber 2 (ADC2)
Drying Air drying
Detection
  1. 366 nm after derivatisation with 1% aluminium chloride in ethanol.

High Performance Liquid Chromatography (HPLC)

Test solution Weigh about 1.0 g of P. macrocarpa dried leaves powder of 0.355 mm size in a 50 mL conical flask. Add 5 mL of distilled water and soak the mixture for 30 min at room temperature. Add 20 mL of petroleum ether (b.p 40-60oC) and sonicate the mixture for 15 min at room temperature (cleanup process). Discard the petroleum ether and repeat the cleanup process twice. Allow the raw material to stand for 1 hour at room temperature. Add 10 mL of ethanol and sonicate the mixture for 15 min at room temperature. Centrifuge the mixture and use the supernatant as test solution.
Standard solution Dissolve mangiferin standard [CAS no: 4773-96-0] in ethanol to produce a standard concentration 0.2 mg/mL.
Chromatographic system

Detector: 370 nm

Column: 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: 10 µL

Mobile Phase (gradient mode)

Run Time

(min)

A – Formic acid (0.1% in water)
 (%)

B – Formic acid (0.1% in methanol)
(%)

0.0

85

15

2.0

75

25

15.0

75

25

30.0

0

100

35.0

0

100

35.1

85

15

40.0

85

15

System suitability requirement

Perform at least five replicate injections of the standard solution (0.2 mg/mL). The requirements of the system suitability parameters are as follow:

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

(a)

4b

(b)

Figure 4 : Whole HPLC chromatogram of (a) mangiferin standard solution (0.2 mg/mL) at tr= 17.5 min and (b) ethanol extract of Phaleria macrocarpa dried leaves powder showing peak corresponding to mangiferin standard solution at tr= 17.5 min.

5a

(a)

5b

(b)

Figure 5 : HPLC chromatogram highlighting the elution region of mangiferin in (a) mangiferin standard solution (0.2 mg/mL) at tr= 17.5 min and (b) ethanol extract of Phaleria macrocarpa dried leaves powder showing peak corresponding to mangiferin standard solution at tr= 17.5 min.

6

Figure 6 : UV spectrum of mangiferin standard solution (0.2 mg/mL) and ethanol extract of Phaleria macrocarpa dried leaves powder.

PURITY TESTS

The purity tests are based on P. macrocarpa dried leaves 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 2%
Loss on Drying
Not more than 9%
Extractive Values
Water-soluble extracts
Hot method Not less than 27%
Cold method Not less than 22%
Ethanol-soluble extracts
Hot method Not less than 19%
Cold method Not less than 15%

SAFETY TESTS

The safety tests are based on P. macrocarpa dried leaves 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 P. macrocarpa dried leaves was found to contain benzophenoic glucoside (4,5-dihydroxy, 4’-methoxybenzophenone-3-O-β-D-glucoside; phalerin) [ 6 ].

Ethyl acetate extract of P. macrocarpa dried leaves was found to contain benzophenone aglucon (2,6,4’-trihydroxy-4-methoxybenzophenone) [ 7 ].

n-hexane extract of P. macrocarpa dried leaves was found to contain sterol (β-sitosterol and stigmasterol) [ 8 ].

MEDICINAL USES

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

Traditionally, the leaves of P. macrocarpa is used for intestinal infection and allergy [ 9 ].

Biological and pharmacological activities supported by experimental data

Antioxidant activity
2,6,4’-trihydroxy-4-methoxybenzophenone isolated from ethyl acetate extract of P. macrocarpa dried leaves (10 – 18 µg/mL) showed DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity with 50% inhibition concentration (IC50) value of 10.57 ± 0.72 µg/mL compared to quercetin (1.2-3.6 µg/mL) with IC50 value of 2.93 ± 2.12 µg/mL) [ 7 ].

Anti-hypercholesterolemic activity
Ethyl acetate extract of P. macrocarpa dried leaves (12.5 µg/mL) increased the hepatic scavenger receptor class B type I (SR-BI) expression to reduce cholesterol levels by 95% compared to positive control rosiglitazone (0.04 µM; 100%) [ 10 ].

Active fraction (CF6) of ethyl acetate extract of P. macrocarpa dried leaves (12.5 µg/mL) increased the SR-BI expression to reduce cholesterol levels by 95% compared to positive control rosiglitazone (0.04 µM; 100%) [ 10 ].

Ethly acetate extract of P. macrocarpa dried powder leaves (0.5 g/kg) was administered orally to female hypercholesterolemia-induced Sprague Dawley rats (7 weeks old; 150 – 180 g) once daily for a duration of 28 days. The extract showed significant (p < 0.05) reduction in the total cholesterol level by 224.9% and increased the HDL cholesterol level by 157% compared to cholesterol treated group [ 10 ].

Clinical studies

Information and data have not been established. 

SAFETY INFORMATION

Preclinical studies (Toxicology studies)

Acute Toxicity
Oral single dose acute toxicity study on female Sprague Dawley rats (aged between 8 and 12 weeks old) using aqueous extract of P. macrocarpa leaves showed no toxic effect on the parameters observed, including behaviour, 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 2,000 mg/kg body weight [ 11 ].

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

  1. The Plant List [homepage on the Internet]. Phaleria macrocarpa (Scheff.) Boerl, 2012 [cited on 7 April 2016]. Available from: http://www.theplantlist.org/tpl1.1/record/tro-50315226
  2. Anggraini T, Lewandowsky P. The exotic plants of Indonesia: Mahkota dewa (Phaleria macrocarpa), sikaduduak (Melastoma malabathricum Linn) and mengkudu (Morinda citrifolia) as potent antioxidant sources. International Journal on Advanced Science Engineering Information Technology. 2015;5(2):115-118.
  3. Hai Kou Yi Yu Cao Mu Health Food Co. Ltd [Internet] Phaleria macrocarpa; 2015 [cited on 20 December 2016]. Available from: http://www.hkyycm.com/news2.aspx?cid=1&id=21
  4. Rabia A, Mohammad ZBA, Aidiahmad D, Amirin S, Muhammad IU. Phytochemistry and medicinal properties of Phaleria macrocarpa (Scheff.) Boerl. extracts. Pharmacognosy Review. 2013; 7(13):73-80.
  5. Hasimah A, Nor Nafizah MN, Norhayati D, Hasnah I. The inflorescence and infructescence morphology of Phaleria macrocarpa (Boerl.) Scheff. Journal of Science and Mathematics. 2013;5:67-72.
  6. Hartati WMS, Mubarika S, Gandjar I, Hamann MT, Rao KV, Wahyuono S. Phalerin, a new benzophenoic glucoside isolated from the methanolic extract of Mahkota Dewa [Phaleria macrocarpa (Scheff).Boerl.] leaves. Majalah Farmasi Indonesia. 2005;16(1):51-57.
  7. Susilawati, Matsjeh S, Pranowo HD, Anwar C. Antioxidant activity of 2,6,4’-trihydroxy-4-methoxybenzophenone from ethyl acetate extract of leaves of mahkota dewa (Phaleria macrocarpa (Scheff.) Boerl.). Indonesian Journal of Chemistry. 2011;11(2):180-185.
  8. Othman SNAM, Sarker SD, Talukdar AD, Ningthoujam SS, Khamis S, Basar N. Chemical constituents and antibacterial activity of Phaleria macrocarpa (Scheff.) Boerl. International Journal of Pharmaceutical Sciences and Research. 2014; 5(8):3157-3162.
  9. Harmanto N. Conquering disease in unison with Mahkota dewa. North Jakarta: Ir. Harmantop, 2003; p.14.
  10. Yosie A, Sifzizul TMT, Habsah M, Jasnizat S, Desy FS, Guat SC, Effendy MAW. Phaleria macrocarpa Boerl. (Thymelaeaceae) leaves increase SR-BI xpression and reduce cholesterol levels in rats fed a high cholesterol diet. Molecules. 2015; 20:4410-4429.
  11. Norzahirah A, Umi Rubiah SZ, Bazilah J, Nurul Syahira S, Dalnitha Lea N, Tavinraj R, Teh BP. Acute oral toxicity study of selected Malaysian medicinal herbs on Sprague Dawley rats. Institute for Medical Research, Ministry of Health ; 2019. Reportno.: NON-GLP/2019/07/01.