Marantodes pumilum (Blume) Kuntze var. pumila (Roots)

MALAYSIAN HERBAL MONOGRAPH

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Kacip Fatimah Variety Pumila Roots

Marantodes pumilum (Blume) Kuntze var. pumila

(syn. Labisia pumila (Blume) Fern.-Vill. var. pumila)

Primulaceae

 

                                fig1a
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 fig1b fig1c
(b) (c)
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Figure 1 : Marantodes pumilum var. pumila. (a) Whole plant (b) leaf petioles and flower; (c) roots; (d) fruits  (Photos courtesy of Hawa ZE Jaafar, Universiti Putra Malaysia, 2017)  

   

DEFINITION

Kacip fatimah variety pumila roots consist of the powder of dried stems and roots of Marantodes pumilum (Blume) Kuntze var. pumila (Primulaceae). This variety can be physically differentiated from the variety of M. pumilum var. alata based on the morphology of leaf petiole. M. pumilium var. pumila has slightly winged petiole, whereas M. pumilum var. alata has rather broadly winged petiole. [1]

SYNONYM

Ardisia pumila Blume,Ardisia pumila Blume α genuiana Scheff., Labisia pumila (Blume) Fern.-Vill. var. pumila, Labisia pumila (Blume) Fern.-Vill. α genuiana Mez, Labisia pumila (Blume) Fern.-Vill. forma genuiana Mez ex Airy Shaw, Labisia punctata (Reinw.) Lindl. forma pumila (Blume) Airy Shaw [1, 2]. 

VERNACULAR NAMES

Fatimah’s childbirth medicine and Fatimah’s betel scissors (English); Kacip fatimah, hati fatimah, selusuh fatimah, akar fatimah, kunci fatimah, pokok pinggang, rumput siti fatimah, rumput palis, tadah matahari, mata pelanduk rimba and bunga belangkas hutan (Malay). [3, 4]

CHARACTER

Colour            : Light brown

Odour             : Slight odour

Taste              : Tasteless

IDENTIFICATION

Plant Morphology

Marantodes pumilum var. pumila is a slow growing erect, 10–40 cm high or prostrate up to 60 cm long or longer, decumbent or creeping undershrubs.  Stems erect, green to dark brown in colour with distinct scars of fallen leaves;  nodes of the stem are capable of rooting and from the older nodes. Leaves simple more than 5 per plant, spirally arranged, elliptic or ovate; lamina (narrowly or broadly) elliptic or ovate 10-30 cm long, 1.3–11 cm wide, base acute or obtuse, more or less decurrent, or sometimes slightly cordate, margin serrate, dentate, crenulate, subentire or entire, apex acuminate, acute or obtuse, midrib narrowly impressed above, secondary nerves anastomosing at 1–4 mm from margin; petioles slightly winged, 4–5 cm long. Inflorescence axillary, panicles or racemes, with corymb-like fascicles of 2–9 flowers, 5–25 cm long; corymb and floral bracts narrowly triangular, 1–3 (–5) mm long; pedicels 1–3 mm long. Flowers white or pinkish white, hermaphrodite, 5-merous, zygomorphic, glandular pusticulate, glabrous; sepals 5, basally connate for ⅓–½ of its length, segments valvate, lobes triangular 0.5–1 mm long, acute to acuminate; petals 5 basally connate near the base, segments valvate, lobes ovate, 2–2.5 by 1–1.5 mm, apex subacute; in mature buds, just before anthesis subglobose, 1.5–2 mm long, top obtuse; stamens 5, 1–1.5 mm long, epipetalous, enclosed by the petals; filaments short, c. 0.2 mm long, connective appendages 0.1–0.3 mm long, backside glandular pusticulate; anthers sagittate, about 1.2 mm long; ovary superior, subglobose; placenta subglobose; ovules 6–7, uniseriate; style subulate; stigma small, notched. Fruit drupaceous, subglobose, 4–5 mm in diameter. Seed 1, subglobose, ribbed. Roots can be seen protruding out. [1, 5]

Microscopy

Powdered material consists of elongated parenchyma cells; elongated brachysclereids; vessels with spiral thickening; scale trichomes; prism and druse calcium oxalate crystals; cork cells; round epidermal cells; simple multicellular trichomes; abundant starch grains that are mainly elongated, oval or irregular in shape; absence of fusiform and rectangular brachysclereids [6].

 

fig2a fig2b fig2c
(a) (b) (c)
 fig2d fig2e  fig2f 
(d) (e) (f)
 fig2g  fig2h fig2i 
(g) (h) (i)
 fig2j  fig2k  fig2l
(j) (k) (l)
Figure 2 : Microscopic characters of Marantodes pumilum var. pumila roots powder of 0.106 mm particle size. (a) Elongated parenchyma cells (magnification 100x); (b) elongated brachysclereid (magnification 100x); (c) vessel with spiral thickening (magnification 400x); (d) peltate trichome (magnification 100x); (e) prism calcium oxalate crystal (magnification 400x); (f) druse calcium oxalate crystal (magnification 400x); (g) cork cells (magnification 100x); (h) round epidermal cells  (magnification 100x); (i) simple multicellular trichome (magnification 100x); (j) cluster of starch grains in parenchyma cell (magnification 100x); (k) starch grains (magnification 100x); (l) starch grains (magnification 400x). [Scale bars: a, b, d, g, h, i, j, k = 100 µm; c, e, f, l = 20 µm]

 

Chemical Tests 

Observation of solution after treatment with various reagents:

 

Test for the presence of steroids : Bluish green
Test for the presence of saponins : Formation of stable foam for 30 minutes
Test for presence of phenolic (tannins) : Bluish green

Thin Layer Chromatography (TLC)

Test solution : Weigh about 4.0 g of M. pumilum var. pumila dried roots powder of 0.106 mm size in a 250 mL round bottom flask and add 15 mL of methanol into the flask. Reflux the sample for 1 hr and allow to cool. Filter the mixture and use the filtrate as the test solution.
Standard solution : Dissolve gallic acid standard (CAS no.: 149-91-7) in methanol to produce a standard solution of 0.1 mg/mL.
Stationary phase : HPTLC Glass Silica Gel 60 F254, 20 x 10 cm
Mobile phase : Toluene : ethyl acetate : formic acid; (5 : 5 : 1) (v/v/v)
Application :
  1. Gallic acid standard solution (S); 10 µL, 8 mm as a band
  2. Methanol extract of M. pumilum var. pumila dried roots powder (L); 10 µL, 8 mm as a band
Development distance : 7 cm, manual chamber
Drying : Air drying
Detection :

(a) visible light;

(b) UV 254 nm;

(c) UV 366 nm.

 

fig3
Figure 3 : TLC chromatogram of gallic acid (S) and methanol extract of Marantodes pumilum var. pumila dried roots powder (L) under (a) visible light, (b) UV at 254 nm, (c) UV at 366 nm.

High Performance Liquid Chromatography (HPLC)

Test solution : Weigh about 5.0 g of M. pumilum var. pumila dried roots powder of 0.106 mm size into a flask and add 100 mL of absolute ethanol. Reflux the mixture for 30 min. Filter the mixture through a filter paper. Evaporate to dryness using a rotary evaporator. Then reconstitute the residue with 10 mL of methanol (HPLC grade). Sonicate the mixture for 15 min and then centrifuge at 2000 rpm for 15 min. Filter the supernatant through a 0.45 μm syringe filter and inject into HPLC column.
Standard solution : Dissolve gallic acid standard (CAS no.: 149-91-7) in methanol to produce a standard solution of 0.1 mg/mL. 
Chromatographic system  :

Detector: UV 270 nm

Column: C18 column (5.0 µm, 4.6 mm I.D x 150 mm)

Column oven temperature: 30 ºC

Flow rate: 1 mL/min

Injection volume: 1 µL
Mobile phase (Isocratic mode) : 3% phosphoric acid (85%) buffer : acetonitrile; (9:1) (v/v)
Run time : 10 min
System suitability requirements :

Perform at least five replicate injections of the gallic acid standard solution (0.1 mg/mL). The requirements of the system suitability parameters are as follow:

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

  

 fig4a
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 fig4b
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Figure 4 : Whole HPLC chromatogram of (a) gallic acid standard solution (0.1 mg/mL) at tr= 2.767 min and (b) ethanol extract of Marantodes pumilum var. pumila dried roots powder showing peak corresponding to gallic acid standard solution at tr= 2.769 min.

  

 fig5a
(a)
 fig5b
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Figure 5 : HPLC chromatogram highlighting the elution region of gallic acid in (a) gallic acid standard solution (0.1 mg/mL) at tr = 2.767 min and (b) ethanol extract of Marantodes pumilum var. pumila dried roots powder showing peak corresponding to gallic acid standard solution at tr = 2.769 min.

 

 fig6
Figure 6 : UV spectrum of gallic acid standard solution (0.1 mg/mL) and ethanol extract of Marantodes pumilum var. pumila dried roots powder.

PURITY TESTS

The purity tests, except foreign matter test, are based on M. pumilum var. pumila dried roots powder of 0.106  mm particle size.

 

Foreign Matter
Not more than 2%

 

Ash Contents
Total ash : Not more than 9%
Acid-insoluble ash : Not more than 1%
Water-soluble ash : Not less than 1%

 

Loss on Drying
Not more than 10%

 

Extractive Values
Water-soluble extracts
Hot Method : Not less than 13%
Cold Method : Not less than 6%
Ethanol-soluble extracts
Hot Method : Not less than 6%
Cold Method : Not less than 4%

SAFETY TESTS

The safety tests are based on M. pumilum var. pumila dried roots powder of 0.106  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 10cfu/g
 Total yeast and mould count  :   Not more than 10cfu/g
 Bile-tolerant gram negative bacteria  :   Not more than 10cfu/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 the freeze-dried roots of M. pumilum var. pumila was found to contain flavonoids (eg. kaempferol, myricetin, rutin, apigenin, daidzein, genistein and naringin) and phenolics (eg. gallic acid, caffeic acid, pyrogallol). [7] 

MEDICINAL USES

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

In the Malay peninsula, a decoction of the roots or plant Labisia pumila (M. pumilum) is traditionally used to induce and expedite labor, as well as a remedy for postpartum protective medicine, menstrual problem, flatulence, intestinal infection and ‘sickness in the bones’. The roots are also traditionally used for meroyan. [3, 8, 9]

Biological and pharmacological activities supported by experimental data

Antioxidant activity

Methanol extract of M. pumilum var. pumila roots had IC50 values of > 500 μg/mL for both DPPH free radical scavenging and FRAP activities; the same as methanol extract of M. pumilum var. alata roots (DPPH and FRAP: IC50 > 500 μg/mL), compared to BHT and α-tocopherol standards with IC50 values of 78.752 and 36.031 µg/mL, respectively, for DPPH assay; and 89.76 and 61.86 µg/mL, respectively, for FRAP assay. The results suggested that the roots had low antioxidant activity. [10]

Antimicrobial activity

Methanol extracts of M. pumilum var. pumila and M. pumilum var. alata roots (300 µg/disc) displayed inhibitory activity against Gram-positive bacteria using a disc diffusion method: Bacillus subtilis (pumila = 0.99 mm and alata = 0.94 mm compared to kanamycin = 1.25 mm), Bacillus cereus (pumila = 0.96 mm and alata = 0.85 mm compared to kanamycin = 1.28 mm), Staphylococcus aureus(pumila = 0.65 mm and alata = 0.39 mm compared to kanamycin = 1.11 mm) and Micrococcus luteus (pumila = 0.41 mm and alata = 0.39 mm compared to kanamycin = 0.88 mm); as well as against Gram-negative bacteria: Escherichia coli (pumila = 0.93 mm and alata = 1.01 mm compared to kanamycin = 1.48 mm), Pseudomonas aeruginosa (pumila = 0.47 mm and alata = 0.47 mm compared to kanamycin = 1.03 mm), Enterobacter aerogenes (pumila = 0.98 mm and alata = 1.11 mm compared to kanamycin = 1.33 mm) and Klebsiella pneumonia (pumila = 0.62 mm and alata = 0.83 mm compared to kanamycin = 1.31 mm). [11]

Methanol extracts of M. pumilum var. pumila and M. pumilum var. alata roots (500 µg/well) also showed inhibitory activity against pathogenic fungi using an agar well diffusion assay: Fusarium sp. (pumila = 0.58 cm and alata = 0.59 cm compared to amphotericin B = 1.41 cm), Candida sp. (pumila = 0.54 cm and alata = 0.65 cm compared to amphotericin B = 1.43 cm) and Mucor sp. (pumila = 0.45 cm and alata = 0.43 cm compared to amphotericin B = 0.90 cm). [11]

Anti-inflammatory activity

Methanol extract of M. pumilum var. pumila roots (100 µg/mL) inhibited LPS/IFN-γ-induced nitric oxide (NO) production in RAW 264.7 cell lines moderately at 66.45%, whereas M. pumilum var. alata roots (100 µg/mL) had strong inhibition (75.68%), compared to N-nitro-L-arginine methyl esther (L-NAME) with an inhibition value of 78.43%. [12]

Phytoestrogenic activity

Water and ethanol extracts M. pumilum var. pumila roots (1 – 1000 μg/mL) did not show oestrogenic activity when tested using human endometrial adenocarcinoma cells of the Ishikawa-Var I line in vitro based on oestrogen-specific enhancement of alkaline phosphatase activity. [13] 

Cytotoxic activity

Methanol extract of M. pumilum var. pumila roots was found to inhibit human cancer MCF-7 and MDA-MB-231 cell lines with IC50 values of 81.92 and 98.93 μg/mL, respectively; while that for M. pumilum var. alata roots were 86.14 and 110.30 μg/mL, respectively, compared to the positive standard tamoxifen (IC50 = 36.54 and 35.46 μg/mL, respectively). The extracts were also found to weakly inhibit Chang liver cell lines with IC50 values of 192.16 and 160.90 μg/mL, respectively, compared to tamoxifen (IC50 = 34.97 μg/mL). [12]

Clinical studies

Information and data have not been established.

SAFETY INFORMATION

Preclinical studies (Toxicology studies)

Information and data have not been established.

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. Sunarno B. Revision of the genus Labisia (Myrsinaceae). Blumea 2005;50: 579–597.
  2. The Plant List 2013. [Internet] Marantodes pumilum (Blume) Kuntze; 2012 [cited on 26th August 2016]. Available from: http://www.theplantlist.org/tpl1.1/record/tro-100363713
  3. Burkill, I.H. A Dictionary of the Economics Products of the Malay Peninsular. 2nd Ed. Kuala Lumpur: Ministry of Agriculture and Cooperative. 1966.
  4. Forestry Department Peninsular Malaysia. [Internet] Labisia pumila Benth; 2016 [cited on 26th August 2016]. Available from: http://www.forestry.gov.my/herba/kacipfatimah_en.pdf
  5. Stone BC. Notes on the genus Labisia Lindl. (Myrsinaceae). Malayan Nature Journal. 1988;42:43-51.
  6. Nor-Ashila A, Jamal JA, Noraini T, Nur Ain MH, Mohd Ruzi AR, Carla WS, Kartiniwati M, Khairana H, Juriyati J. Comparative study of three Marantodes pumilum varieties by microscopy, spectroscopy and chromatography. Revista Brasileira de Farmacognosia. 2016;26:1-14.
  7. Ehsan K, Hawa ZEJ. HPLC and GC-MS determination of bioactive compounds in microwave obtained extracts of three varieties of Labisia pumila Benth. Molecules 2011;16:6791-6805.
  8. Perry LM, Metzger J. Medicinal Plants of East and Southeast Asia : Attributed Properties and Uses. Massachusetts: MIT Press. 1985;282.
  9. Kamarudin MS, Latiff A. Tumbuhan Ubatan Malaysia. Bangi: Pusat Pengurusan Penyelidikan Universiti Kebangsaan Malaysia. 2002;273.
  10. Karimi E, Hawa ZEJ, Sahida A. Phenolics and flavonoids profiling and antioxidant activity of three varieties of Malaysian indigenous medicinal herb Labisia pumila Benth. Journal of Medicinal Plants Research. 2011;5(7):1200-1206.
  11. Karimi E, Hawa ZEJ, Sahida A. Phytochemical analysis and antimicrobial activities of methanolic extracts of leaf, stem and root from different varieties of Labisia pumila Benth. Molecules 2011;16:4438-4450.
  12. Karimi E, Hawa ZEJ, Sahida A. Antifungal, anti-inflammatory and cytotoxicity activities of three varieties of Labisia pumila Benth: from microwave obtained extracts. BMC Complement Alternative Medicine. 2013;13:20-29.
  13. Jamal JA, Houghton PJ, Milligan SR, Ibrahim J. The oestrogenic and cytotoxic effects of the extracts of Labisia pumila var. alata and Labisia pumila var. pumila in vitro. Jurnal Sains Kesihatan Malaysia. 2003;1:53-60.