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Home > Medicinal Herbs and Plant Database > Malaysian Herbal Monograph > Zingiber montanum (J.Koenig) Link ex A.Dietr.
Malaysian Herbal Monograph
  • Written by Shahrul
  • Updated on November 7, 2022

Bongelai Rhizome

Zingiber montanum (J.Koenig) Link ex A.Dietr.

Zingiberaceae

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Figure 1 : Zingiber montanum. (a) Whole plant; (b) pseudostem and rhizome; (c) alternate distichous leaves; (d) rhizome. (Photos courtesy of Mohd Fauzi Sayuti, UKM, 2015)

DEFINITION

Bongelai rhizome consists of the powder of dried rhizome of Zingiber montanum (J.Koenig) Link ex A.Dietr. (Zingiberaceae) [ 1 ].

SYNONYM

Amomum cassumunar (Roxb.) Donn, Amomum montanum J.Koenig, Amomum xanthorhiza Roxb. Ex Steud., Cassumunar roxburghii Colla, Jaegera montana (J.Koenig) Giseke, Zingiber anthorrhiza Horan., Zingiber cassumunar Roxb., Zingiber cassumunar var. palamauense Haines, Zingiber cassumunar var. subglabrum Thwaites, Zingiber cliffordiae Andrews, Zingiber luridum Salisb., Zingiber montanum (J. König ex Retz.) Theilade, Zingiber purpureum Roscoe, Zingiber purpureum var. palamauense (Haines) K.K.Khanna and Zingiber xantorrhizon Steud [ 2 ].

VERNACULAR NAMES

Cassumunar ginger (English) [ 3 ]; bongelai (Malay) [ 1 ]; ye jiang (Chinese); kattu ingi (Tamil) [ 4 ].

CHARACTER

ColourBrown [ 1 ]
OdourStrong and sharp [ 1 ]
TasteHot and sharp [ 1 ]

IDENTIFICATION

Plant Morphology

Z. montanum is a perennial and clumping herb; pseudostem erect, cylindrical, enveloped by leafy sheaths and reaching 1.2–1.8 m high. Rhizomes horizontal, creeping, cylindrical to ovoid, tuberous, irregular, profusely branched and palmately, laterally compressed and strongly aromatic with yellow flesh colour. Leaves simple, distichous, alternate, subsessile or shortly petiolate, lanceolate-oblong and 3.5–5.5 × 18–35 cm; leaf sheaths are oblong, with membranous margins; ligules are ovate and membranous. Inflorescence radical; spikes are cylindrical, fusiform or cone like, borne on a peduncle spike (scape) arising from rhizome, 8–60 cm high with 5–7 cataphylls; bracts divided into outer and inner, spirally arranged, very dense, persistent and red or purplish brown; the outer is broadly ovate to suborbicular and cucullate, while the inner is ovate and glabrous. Flowers bisexual, ebracteolate, zygomorphic and epigynous; calyx is 1.2–1.5 cm, membranous, glabrous and white; corolla has 4 lateral lobes and linear-lanceolate, yellowish-white and reddish lineolate on margins; labellum is white or pale yellow and suborbicular; apex is emarginate; central lobe is broadly rounded; stamen 1 cm long and pale yellow; ovary is 3 loculed, inferior, 3–4 mm long and pubescent. Capsule is ovoid to subglobose and 1–1.5 cm diameter [ 4 ].

Microscopy

Powdered material consists of starch granules that are oval and tear-drop in shape; lignified polygonal parenchyma cells; scalariform vessels; stellate trichomes; fibres, oil globules and prism calcium oxalate crystals [ 1 ].

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Figure 2 :Microscopic characters of Zingiber montanum dried rhizome powder of 0.106 mm size. (a) Oval shaped starch granule (magnification 100×); (b) tear drop shaped starch granules stained blue-black (magnification 100×); (c) lignified polygonal parenchyma (magnification 100×); (d) fragment of scalariform vessel (magnification 100×); (e) stellate trichome (magnification 100×); (f) oil globules stained orange-pink (magnification 100×); (g-h) prism calcium oxalate crystals (magnification 400×). [Scale bars: a-f = 100 µm; g-h = 20 µm]

Colour Tests

Observed colour of solution after treatment with the following reagents:

H2SO4 (conc.)Brown
NaOH (5%)Orange

Thin Layer Chromatography (TLC)

Figure 3 : TLC profiles of curcumin standard (S), ethanol extract of Zingiber montanum dried rhizome powder (L) observed under (a) visible light, (b) UV at 254 nm and (c) UV at 366 nm.

Test Solutions Weigh about 5.0 g of Z. montanum dried rhizome powder of 0.106 mm particle size in a 250 mL round bottom flask. Add 100 mL of absolute ethanol and reflux the mixture for 30 min at 60°C. Filter the mixture with filter paper and evaporate the filtrate to dryness using a rotary evaporator. Reconstitute the residue with 10 mL methanol and use as the test solution.
Standard solution Dissolve curcumin standard [CAS no: 458-37-7] in methanol to produce 0.3 mg/mL solution.
Stationary Phase HPTLC Glass Silica Gel 60 F254, 10 x 10 cm.
Mobile phase Dichloromethane-methanol, 25:1 (v/v)
Application
  1. Curcumin standard solution (S); 10 µL, 6 mm as a band.
  2. Ethanol extract of Z. montanum dried rhizome powder (L) ; 10 µL, 6 mm as a band.
Development distance 8 cm
Drying Air drying
Detection
  1. Visible light;
  2. UV at 254 nm;
  3. UV at 366 nm.

High Performance Liquid Chromatography (HPLC)

Test solution Weigh about 5.0 g of Z. montanum dried rhizome powder of 0.106 mm particle size in a 250 mL round bottom flask. Add 100 mL of absolute ethanol and reflux the mixture for 30 min at 60°C. Filter the mixture with filter paper and evaporate the filtrate to dryness using a rotary evaporator. Reconstitute the residue with 10 mL methanol. Filter the mixture through a 0.22 µm nylon membrane and use as the test solution.
Standard solution Dissolve curcumin standard [CAS no: 458-37-7] in HPLC grade methanol to produce 0.3 mg/mL solution.
Chromatographic system

Detector: UV 430 nm

Column: C18 column (5.0 µm, 4.6 mm I.D × 250 mm) (Zorbax Exclipse -XDB if necessary)

Column oven temperature: 25°C

Flow rate: 1.0 mL/min

Injection volume: 1 µL
Mobile phase (isocratic mode) 0.1% Orthophosphoric acid in water : acetonitrile; (45 : 55)
Run time 25 min
System suitability requirement

Perform at least five replicate injections of the standard solutions (0.3 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 curcumin standard is not more than 2.0%.
Acceptance criteria
  1. Retention time (tr) of curcumin in the test solution is similar to the tr of the standard solution.
  2. The ultraviolet (UV) spectrum of curcumin in the test solution is similar to the UV spectrum of curcumin in the standard solution (optional supportive data).
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Figure 4 : Whole HPLC chromatogram of (a) curcumin standard solution (0.3 mg/mL) at tr= 7.271 min and (b) ethanol extract of Zingiber montanumdried rhizome powder showing peak corresponding to curcumin standard solution at tr= 7.368 min.

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Figure 5 : HPLC chromatogram highlighting the elution region of curcumin, in (a) curcumin standard solution (0.3 mg/mL) at tr= 7.271 min and (b) ethanol extract of Zingiber montanumdried rhizome powder showing peak corresponding to curcumin standard solution at tr= 7.368 min.

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Figure 6 : UV spectrum of curcumin standard (0.3 mg/mL) and ethanol extract of Zingiber montanum dried rhizome powder.

PURITY TESTS

The purity tests are based on Z. montanum dried rhizome powder of 0.106 mm particle size

Foreign Matter
Not more than 2%
Ash Contents
Total ash Not more than 10%
Acid-insoluble ash Not more than 2%
Loss on Drying
Not more than 14%
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 10%
Cold method Not less than 7%

SAFETY TESTS

The safety tests are based on Z. montanum dried rhizome 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 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

The (70%) methanol extract has been found to contain phenylbutenoids (e.g. zerumbone and (E)-1-(3’,4’-dimethoxyphenyl)but-1-ene) [ 5 ].

The (80%) methanol extract was found to contain phenylbutenoids (e.g. (E)-4-(3’,4’-dimethoxyphenyl)but-3-en-1-ol, (E)-4-(2’,4’,5’-tri-methoxyphenyl)but-3-en-1-ol, (E)-4-(3’,4’,1-trimethoxyphenyl)but-3-en-1-ol [ 6 ].

Methanol extract of the fresh rhizomes was found to contain phenylbutanoids (e.g cassumunols A-H, cassumunaquinones 1 and 2, 3,4-dimethoxybenzaldehyde, 2,4,5-trimethoxybenzaldehyde,4-(2,4,5-trimethoxyphenyl)-but-1,3-diene, 4-(3,4-dimethoxyphenyl)but-1,3-diene, (Z)-1-(2,4,5-trimethoxyphenyl)buta-1,3-diene, (E)-1-(3,4-dimethoxyphenyl)buta-1,3-diene, (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-ol, (E)-4-(3,4-dimethoxyphenyl)but-3-enyl acetate, (E)-4-(3,4-dimethoxyphenyl)but-3-ene-1,2-diol, (E)-1-(3,4-dimethoxyphenyl)but-1-ene, (E)-1-(2,4,5-trimethoxyphenyl)but-1-ene, (E)-3-(3,4-dimethoxyphenyl)propenal, (E)-3-(2,4,5-trimethoxyphenyl)propenal, (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-ol, (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-yl acetate and zerumbone); phenylbutenoid dimers (e.g (±)-trans-3-(3,4-dimethoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohex-1-ene, (±)-trans-3-(4-hydroxy-3-methoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohex-1-ene,   (±)-trans-3-(2,4,5-trimethoxyphenyl)-4-[(E)-2,4,5-trimethoxystyryl]cyclohex-1-ene,   (±)-cis-3-(3,4-dimethoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohex-1-ene, (±)-cis-3-(2,4,5-trimethoxyphenyl)-4-[(E)-2,4,5-trimethoxystyryl]cyclohex-1-ene); phenylbutenoid glycoside (e.g. (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-O–β-D-glucopyranoside); conjugated phenylbutanoids (e.g phlains I-VI); curcuminoids (e.g curcumin, cassumunarins A and C) and others (e.g. vanillic acid,   (-)-β-sesquiphellandrene and β-sitosterol) [ 7 , 8 , 9 , 10 , 11 , 12 ].

The ethanol extract has been found to contain phenylbutenoids (e.g. (E)-4-(3,4-dimethoxyphenyl)-3-butene-1,2-diol, (E)-4-(3,4-dimethoxyphenyl)-but-3-en-1-ol, (E)-3-(3,4-dimethoxyphenyl)propenal, (E)-4-(3,4-dimethoxyphenyl)-but-3-en-1-yl acetate, (E)-4-(3’,4’-dimethoxyphenyl)but-3-enyl acetate and (E)-4-(3’,4’-dimethoxyphenyl)but-1,3-diene); phenylbutenoid dimers (e.g. (±)-trans-3-(3,4-dimethoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohex-1-ene and (±)-cis-3-(3,4-dimethoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohex-1-ene) [ 13 , 14 ].

The acetone extract was found to contain phenylbutenoid monomers (e.g (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-ol, (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-yl acetate, (E)-3-hydroxy-1-(3,4-dimethoxyphenyl)but-1-ene, (E)-4-(4-hydroxy-3-methoxyphenyl)-but-3-en-1-yl acetate, (E)-4-(4-hydroxy-3-methoxyphenyl)but-2-en-1-ol, (E)-2-hydroxy-4-(3,4-dimethoxyphenyl)but-3-en-1-ol and (E)-2-methoxy-4-(3,4-dimethoxyphenyl)but-3-en-1-ol; phenylbutenoid dimers (e.g. (±)-cis-1,2-bis[(E)-3,4-dimethoxystyryl]cyclobutane, (±)-cis-3-(3,4-dimethoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohexene, (±)-trans-3-(3,4-dimethoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohexene and (±)-cis-3-(2,4,5-dimethoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohexene); curcuminoids (e.g cassumunins A-C and cassumunarins A-C) [ 15 , 16 , 17 , 18 , 19 ].

The chloroform extract has been found to contain phenylbutanoids (e.g cis-3-(3,4-dimethoxyphenyl)-4-[(E)-3,4-dimethyoxystyryl]cyclohex-1-ene, cis-3-(2,4,5-trimethoxyphenyl)-4-[(E)-2,4,5-trimethyoxystyryl]cyclohex-1-ene, cis-3-(3,4-dimethoxyphenyl)-4-[(E)-2,4,5-trimethyoxystyryl]cyclohex-1-ene, cis-4-[(E)-3,4-dimethoxystyryl]-3-(2,4,5-trimethoxyphenyl)cyclohex-1-ene, trans-3-[3,4-dimethoxyphenyl]-4-[(E)-3,4-dimethoxystyryl]cyclohex-1-ene, trans-3-[3,4-dimethoxyphenyl]-4-[(E)-2,4,5-trimethoxystyryl]cyclohex-1-ene, (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-ol, (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-yl palmitate, (E)-1-(3,4-dimethoxyphenyl)but-1-ene, (E)-1-(2,4,5-trimethoxyphenyl)but-1-ene and (E)-1-(3,4-dimethoxyphenyl)butadiene); phenylnapthoquinones (e.g. 8-(3,4-dimethoxyphenyl)-2-methoxynaphto-1,4-quinone and 2-methoxy-8-(2,4,5-trimethoxyphenyl)naptho-1,4-quinone); and others (e.g. vanillin, vanillic acid, veratric acid, terpinen-4-ol and curcumin) [ 20 ].

The hexane extract has been found to contain phenylbutenoids (e.g. (E)-4-(3′,4′-dimethoxyphenyl)but-3-enyl acetate, cis-3-(3′,4′-dimethoxyphenyl)-4-[(E)-3″‘,4″‘-dimethoxystyryl]cyclohex-1-ene, cis-3-(3′,4′-dimethoxyphenyl)-4-[(E)-2”’,4”’,5”’-trimethoxystyryljcyclohex-1-ene, cis-3-(2′,4′,5’trimethoxyphenyl)-4-[(E)-2”’,4”’,5”’- trimethoxystyryl]cyclohex-1-ene, (E)-4-(3′,4′-dimethoxyphenyl)but-3-en-1-ol, (E)-4-(3′,4′-dimethoxyphenyl)but-3-en-2-ol, (E)-4-(3’,4’-dimethoxyphenyl)but-3-en-l-y1 palmitate,4-(3’,4’-dimethoxyphenyl)but-1,3-diene, 4-(3’,4’-dimethoxyphenyl)but-3-ene, 4-(2’,4’,5’-trimethoxyphenyl)but-1,3-diene and 4-(2’,4’,5’-trimethoxyphenyl)but-3-ene); benzaldehydes (e.g. 3,4-dimethoxybenzaldehyde and 2,4,5-trimethoxybenzaldehyde); and phenylnapthoquinone (e.g. 8-(3′,4′-dimethoxypheny1)-2-methoxynaphtho-1,4-quinone) [ 21 , 22 , 23 , 24 ].

Essential oil of the fresh rhizomes was found to contain monoterpenoids (e.g. terpinen-4-ol, α-terpineol, terpinolene, terpineneol, α-pinene, β-pinene, (Z)-ocimene, allo-ocimene, neo-allo-ocimene, 3-carene, 2-carene, δ-carene, α-terpinene, γ-terpinene, m-cymene, o-cymene, ρ-cymene, ρ-cymen-8-ol, α-thujene, β-myrcene, α-phellandrene, β-phellandrene, sesquiphellandrene, β-sesquiphellandrene, linalol, trans-linaool oxide, trans-piperitol, cis-piperitol, borneol, methyleugenol, bornyl acetate, eucalyptol, D-limonene, limonene and 3,4-dimethoxycinnamaldehyde); sesquiterpenoids (e.g. germacrene D, γ-selinene, α-senene, α-bergamotene, α-caryophyllene, caryophyllene, caryophyllene oxide, β-bisabolene, zingiberene, juniper camphor, camphor, δ-cadinene, δ-elemene, 2,6,9,9-tetramethyl-2,6,10-cycloundecatrien-1-one, sabinene, sabinene hydrate, trans-sabinene hydrate, cis-sabinene hydrate, (Z)-sabinene hydrate, camphene, γ-gurjunenepoxide-(1), 4-terpinyl acetate, terpinyl acetate, (Z)-ρ-menth-2-en-1-ol and myrtenol); phenylbutanoid dimers (e.g. trans-1-(3,4-dimethoxyphenyl)but-1-ene, trans-1-(3,4-dimethoxyphenyI)butadiene, trans-4-(3,4-dimethoxyphenyl)but-3-ene-1-yl acetate, trans-1-(3,6-dimethoxyphenyl)but-1-ene, trans-1-(2,4,5-trimethoxyphenyl)butadiene, trans-4-(3,4-dimethoxyphenyl)but-3-en-1-ol), cis-1-(3,4-dimethoxyphenyl)but-1-ene, cis-1-(3,4-dimethoxyphenyl)butadiene, cis-1-(2,4,5-trimethoxyphenyl)butadiene, cis-4-(3,4-dimethoxyphenyl)but-3-en-1-ol and cis-4-(3,4-dimethoxyphenyl)but-3-en-1-yl acetate); phenylbutanoid monomers (e.g. (E)-1-(3′,4′-dimethoxyphenyl)butadiene, (E)-1-(3,4dimethoxyphenyl)but-1-ene, (E)-1-(3,4-dimethoxyphenyl)buta-1,3-diene, 1-(3,4-dimethoxyphenyl)butane and 1-(2,4,5-trimethoxyphenyl)but-1-ene) and others (e.g 1(10),4-furanodien-6-one, 1,4,7-trimethoxytriquinacene, curzerenone, epicurnerenone, curcumalactone, bornylene, trans-menth-2-en-1-ol, cis-menth-2-en-1-ol, cis-1,2-epoxyterpin-4-ol, vanillin, thuyl alcohol, ρ-mentha-2,4(8)-diene, ar-curcumene, 1,6,10-dodecatrien,7,11-dimethyl-3-methylene (Z), megastigmastriene, triquinacene,1,4-bis(methoxy), 2-propenoic acid, 3(4-methoxyphenyl)ethyl ester, 2-allyl-1,4-dimethoxy-3-methyl benzene, lachnophyllum ester, 1-isopropyl-4-methylbicyclo[3.1.0]hex-2-ene,1-oxaspirol[2.5]octane,5,5-dimethyl-4-(3-methyl-1,3-butadienyl), 1,5,5,8-tetramethyl-12-oxabicyclo[9.1.0]dodeca-3,7-diene, 4,6,6-trimethyl-2-(3-methylbuta-1,3-dienyl)-3-oxatricyclo[5.1.0.0(2,4)]octane, 4-(2,4,5-trimethoxyphenyl)but-1,3-diene and dill ether) [ 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 ].

MEDICINAL USES

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

Traditionally, the rhizome of Z. montanum is used for foot numbness, fever, intestinal disorders, including as a vermifuge in children and for remedy of poisonous plants. Juice of the rhizome is used for boils. The crushed rhizome is spread over the head for relief of headache. The rhizome extract in drink (in form of spirit) is prepared as a lotion for joint pain and bruise. The plant is also applied to the entire body after childbirth. The leaves are boiled and drunk for stomach pain [ 1 , 37 , 38 ].

Biological and pharmacological activities supported by experimental data

Analgesic activity
Methanol extract of Z. montanum rhizome (0.3 and 1 g/kg) was administered to male ddY mice (20 – 25 g) 30 minutes before induction of acetic acid (injected intraperitoneally). The extract inhibited the increase of dye leakage dose-dependently (0.3 g/kg = 304.3 µg), 1 g/kg = 228 µg) compared to indomethacin (219.8 µg) using vascular permeability test
[ 5 ].

Methanol extract of Z. montanum rhizome (1 and 3 g/kg) was administered to male ddY mice (20 – 25 g) 85 minutes before induction of acetic acid (injected intraperitoneally). After 10 minutes, the extract reduced the number of writhing dose-dependently (1 g/kg = 37.3 ± 3.2, 3 g/kg = 21.0 ± 3.1) compared to indomethacin (22.3 ± 4.0) using writhing test
[ 5 ].

Compound (E)-4-(3′,4′-dimethoxyphenyl)but-3-en-2-ol (300 mg/kg) isolated from hexane fraction of Z. montanum was administered to male Swiss albino mice (30 – 40 g) 60 minutes before induction of acetic acid (injected intraperitoneally). The compound showed significant (p < 0.05) reduction in the number of writhing (20 ± 2) compared to control vehicle (40 ± 1) using writhing test [ 22 ].

Phagocytic activity
Phenylbutenoid derivatives, (E)-4-(3’,4’-dimethoxyphenyl)but-3-en-1-ol and (E)-4-(2’,4’,5’-trimethoxyphenyl)but-3-en-1-ol, isolated from the methanol extract of Z. montanum rhizome showed potential immunostimulant property with phagocytosis activity of 99.0 and 90.7%, respectively compared with levamisol hydrochloride (95%) using mouse peritoneum macrophage cells in vitro assay [ 6 ].

Antiulcer effect
Methanol extracts of Z. montanum rhizomes (200 mg/kg and 400 mg/kg) were administered orally to previously fasted 18 h Swiss albino mice, one hour prior to 1N hydrochloric acid (HCl) induced mucosal membrane lesion. The extracts were found to significantly inhibit the lesion by 61.97% at 200 mg/kg and 83.10% at 400 mg/kg doses, compared to omeprazole (30.03% inhibition at 30 mg/kg). Zerumbone isolated from the methanol extract gave 45.77 and 92.25% inhibition at 20 and 40 mg/kg, respectively [ 12 ].

In ethanol (95%) induced gastric mucosal membrane lesions test in mice, zerumbone showed inhibition of 45.35% at a dose of 40 mg/kg, compared with lansoprazole (37.21% inhibition at 30 mg/kg). The compound also exhibited inhibitory effect against indomethacin-induced gastric ulceration in mice with values of 64.76 and 72.38% for 20 and 40 mg/kg, respectively, compared to sucralfate (52.38% inhibiton at 100 mg/kg) [ 12 ].

Antioxidant activity
Cassumunins A, B and C isolated from acetone extract of Z. montanum fresh rhizomes showed stronger antioxidant activity (each 135 μM; 95, 94 and 93%, respectively) than curcumin (135 μM, 78%) as measured by inhibition of autoxidation of linoleic acid in a buffer-ethanol system [ 15 , 17 , 19 ].

Antipyretic activity
(E)-4-(3′,4′-dimethoxyphenyl)but-3-en-2-ol isolated from hexane extract (75, 150 and 300 mg/kg ) of Z. montanum rhizomes was administered orally to yeast-induced hyperthermia in male Sprague Dawley rats (150 – 200 g) exhibited dose-dependent antipyretic effect. The compound rapidly reduced rectal temperature of rats after 30 min of administration at a dose of 300 mg/kg from 39.3 ± 0.1°C to 38.0 ± 0.3°C (p < 0.001), compared to aspirin (300 mg/kg) from 39.4 ± 0.1°C to 38.5 ± 0.2°C (p < 0.001) [ 22 ].

Pancreatic lipase inhibitory activity
Ethanol extract of Z. montanum rhizomes exhibited inhibitory activity on pancreatic lipase (29.17% at 100 ppm) compared with Xenical®/Orlistat as the positive control with inhibition rate of 17.53% at 100 ppm [ 39 ].

Antiproliferative activity
(E)-1-(2,4,5-trimethoxyphenyl)buta-1,3-diene isolated from the methanol extract of Z. montanum rhizomes inhibited invasion of human fibrosarcoma HT 1080 cells through Matrigel-coated filters in vitro with a percentage inhibition of 46.8 ± 7.2% at 30 μM, compared to the positive control, deguelin (57.9 ± 10.5% at 30 μM) [ 7 ].

Phenylbutenoid dimer, cis-3-(3′,4′-dimethoxyphenyl)-4-[(E)-3‴,4‴-dimethoxystyryl]-cyclohex-1-ene, isolated from the petroleum ether extract of Z. montanum rhizomes selectively inhibited proliferation of T-acute lymphoblastic leukaemia CEMss cells with IC50 value of 7.11 ± 0.240 μg/mL compared to the positive control, 5-fluorouracil (IC50 1.54 ± 0.035 μg/mL) [ 40 ].

Phenylbutenoid dimer, (±)-trans-3-(3,4-dimethoxyphenyl)-4-[(E)-3,4-dimethoxy-styryl]cyclohex-1-ene, isolated from the chloroform extract of Z. montanum rhizome was found to inhibit proliferation of A549 human lung cancer cells in a concentration- and time-dependent manner with an IC50 value of 15.6 μM at 72 h compared to ellipticine (0.8 μM at 72 h) by arresting cell cycle progression at the G0/G1 phase by increasing p21 expression and downregulating cyclins and cyclin-dependent kinases [ 41 ].

(±)-trans-3-(3,4-dimethoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohex-1-ene was also found to potently inhibit P-glycoprotein over-expressing multidrug resistant human breast cancer cell line, MCF-7/ADR, by decreasing the IC50 value of the negative control daunomycin (DNM) to 4.31 ± 0.40 μM in the cells (DNM IC50 = 37.1 ± 0.59 μM) compared to verapamil as a positive control that decreased the IC50 of DNM to 6.94 ± 0.40 μM. The compound increased [3H]-DNM accumulation by 395 ± 14.4% compared with that of verapamil (329 ± 18.8%) and reduced P-gp-mediated DNM efflux by 30.8 ± 4.23% more efficiently than verapamil (48.0 ± 5.24%) [ 42 ].

Antimicrobial activity
Essential oil of Z. montanum rhizomes exhibited antimicrobial activity against ringworm Microsporum gypseum with minimum fungicidal concentration (MFC) of 0.31 mg/mL compared to tea tree oil (MFC = 0.25-0.50 mg/mL) and ampicillin (MFC = 0.21 mg/mL) [ 43 ].

Anti-inflammatory activity
Methanol (70%) extract of Z. montanum rhizome suspended in 2% carboxymethyl cellulose was given orally to male ddY mice (20-25 g) 40 mins before the acetic acid-induced vascular permeability test experiment. The extract (1 g/kg) inhibited the dye leakage induced by acetic acid at 228.0 ± 10.9 μg/animal compared to positive control, indomethacin (0.01 g/kg; 219.8 ± 26.3 μg/animal) [ 5 ].

(E)-1-(3,4-Dimethoxyphenyl)butadiene found in the essential oil of Z. montanum rhizomes showed inhibitory effect on carrageenan-induced hind paw edema in male Wistar rats (140-160 g) with 50% inhibition at the dose (ID50) of 3 mg/kg, twice as potent as the reference drug diclofenac (ID50 = 6 mg/kg) [ 44 ].

(E)-4-(3′,4′-dimethoxyphenyl)but-3-en-2-ol isolated from hexane fraction (300 mg/kg) of Z. montanum rhizomes was administered orally to carrageenan-induced hind paw edema in male Sprague Dawley rats (150-200 g). The compound demonstrated 83.9% of edema inhibition. The effect of compound was more potent than aspirin (300 mg/kg; 58.1% inhibition). In carrageenan induced-rat pleurisy, the compound showed inhibition of exudate formation (52.3%), leukocyte accumulation (56.5%) and prostaglandin biosynthesis (32.3%) compared to prednisolone (5 mg/kg; 47.4%, 22.1% and 32.4%, respectively) [ 22 ].

(E)-1-(3,4-dimethoxyphenyl)butadiene isolated from hexane extract of Z. montanum rhizomes was tested for anti-inflammatory effect on ear edema of male Sprague-Dawley rats (50-70 g) induced by ethyl phenylpropiolate, arachidonic acid and 12-O-tetradecanoylphorbol-13-acetate. In ethyl phenylpropiolate-induced edema, the compound exhibited more potent anti-inflammtory effect with an IC50 value of 21 nmol/ear than the standard drug oxyphenbutazone (IC50 = 136 nmol/ear). Similarly in arachidonic acid-induced edema, the compound gave an IC50 value of 60 nmol/ear compared to phenidone (IC50 = 2520 nmol/ear). Whereas in 12-O-tetradecanoylphorbol-13-acetate-induced edema, the compound was 11 times more potent than diclofenac with IC50 values of 660 and 7200 pmol/ear, respectively [ 45 ].

Cassumunins A, B and C isolated from acetone extract of Z. montanum fresh rhizomes were tested for their anti-inflammatory effects using 12-O-tetradecanoylphorbol-13-acetate-induced inflammation method on the ears of male Jcl:ICR mice (6 weeks old). The cassumunins A, B and C (each 0.6 μmol) demonstrated inhibitory effect of 83, 76, and 75%, respectively [ 19 ].

Five compounds isolated from hexane extract of Z. montanum rhizomes were tested for their anti-inflammatory effects using 12-O-tetradecanoylphorbol-13-acetate-induced inflammation method on the ears of male Wistar rats (150-170 g). The compounds (E)-4-(3′,4′-dimethoxyphenyl)but-3enyl acetate, cis-3-(3′,4′-dimethoxyphenyl)4-[(E)-3,‴,4‴-dimethoxystyryl]cyclohex-1-ene, cis-3-(3′,4′-dimethoxyphenyl)-4-[(E)-2‴,4‴,5‴-trimethoxystyryl]cyclohex-1-ene, cis-3-(2′,4′, 5′-trimethoxyphenyl)-4-[(E)-2‴,4‴,5‴-trimethoxystyryl]cyclohex-1-ene, and (E)-4-(3′-4′dimethoxyphenyl)but-3-en-1-ol gave 50% inhibition at the dose (ID50) of 62, 21, 20, 2 and 47 μg/ear, respectively, compared to the reference drug diclofenac (ID50 = 61 μg/ear) [ 21 ].

(E)-1-(3,4-Dimethoxyphenyl)butadiene isolated from hexane extract of Z. montanum rhizomes was tested for inhibitory activities on platelet aggregation induced by collagen, adenosine diphosphate and platelet activating factor in vitro. In collagen-induced platelet aggregation, the compound showed more potent inhibitory activity than the standard drug aspirin with IC50 values of 0.35 and 0.43 mM, respectively. Similarly in the adenosine diphosphate-induced platelet aggregation, the activity of compound and aspirin was rather comparable with IC50 values of 4.85 and 3.98 mM, respectively. Whereas in the platelet activating factor-induced platelet aggregation, the compound was found to be more potent than phenidone with IC50 values of 1.14 and 2.40 mM, respectively [ 45 ].

Anti-asthmatic activity
Ethanol (95%) extract of Z. montanum rhizomes (5-100 μg/mL) was found to significantly inhibited phorbol-12-myristate-13-acetate (200 nmol/L)-induced total mucin production, including MUC2 and MUC5AC genes and proteins expression in the human airway epithelial NCI-H292 cells, compared to phorbol-12-myristate-13-acetate (200 nmol/L) alone (p< 0.05). The potency is comparable to the standard drug budenoside (50 nmol/L). The extract was found to supress phorbol-12-myristate-13-acetate-induced extracellular signal-regulated kinase (ERK) activation in the NCI-H292 cells [ 46 ].

Uterine relaxant effect
(E)-4-(3′,4′-Dimethoxyphenyl)-but-3-en-1-ol isolated from the hexane extract of Z. montanum rhizomes exhibited a dose-related uterine relaxant effect when tested on non-pregnant rat uterus in situ with ED50 of 95.52 mg/kg. It also decreased uterine contraction of isolated rat uterus induced by oxytocin with ED50 of 0.12 mg/mL [ 47 ].

Pediculicidal effect
Crude extract (10% v/v) of Z. montanum rhizomes showed pediculicidal effect against human head lice Pediculus humanus capitis using an immersion method. At 20 sec of exposure, the extract caused 100% mortality compared to the positive control 1.0% w/v malathion (83 ± 12.52%). The median lethal time (LT50) of the exract was 11.30 sec, whereas that of 1.0% w/v malathion was 12.39 sec and the commercial shampoo as a negative control was 87.43 sec [ 48 ].

Laxative effect
Decoction of Z. montanum rhizome (40, 120 and 400 mg/100 g) was administered orally to female Wistar rats (150 – 170 g) for a duration of six hours. The decoction possessed a dose-dependently laxative effect. At the highest dose (400 mg/100 g), the frequency of defecation was 27 times compared to castor oil (23 times of defecation) [ 49 ].

Anthelminthic activity
N–hexane extract of Z. montanum rhizome (189.3 mg/100 mL) showed anthelmintic activity against Ascaris suum by using immersion method. The extract caused 27.8% worm mortality compared to pyrantel pamoate (positive control) [ 50 ].

Anti-allergic activity
Ethanol extract of Z. montanum rhizomes were evaluated for their anti-allergic effect against antigen-induced β-hexosaminidase release on RBL-2H3 cell line. The extract possessed higher activity with IC50 values of 12.9 µg/mL compared to ketotifen fumarate (positive control) with IC50 value of 20.2 µg/mL [ 51 ].

Clinical studies

Information and data have not been established.

SAFETY INFORMATION

Preclinical studies (Toxicology studies)

14-days acute oral toxicity
Granules of Z. montanum rhizome ethanol extract (20% w/w) was used in the acute toxicity study. The granules were prepared by removal of essential oil using steam distillation method prior to extraction of the dried plant residue with ethanol. The ethanol extract was later mixed by wet granulation with excipients. The Z. montanum extract (20% w/w) granules was administered orally as a single dose to male and female Sprague Dawley rats (200 – 280 g). The toxicity effect was observed for 14 days and showed no mortality; however, caused significant (p < 0.05) decrease in body weight of treated male rats, increase in lung and kidney weights in treated female rats and increase in spleen and epididymis weights in treated male rats compared with the control group (LD50 value > 5000 mg/kg) [ 52 ].

Chronic toxicity
Granules of Z. montanum rhizome ethanol extract (20% w/w) was used in the chronic toxicity study. The granules were prepared by removal of essential oil using steam distillation method prior to extraction of the dried plant residue with ethanol. The ethanol extract was later mixed by wet granulation with excipients. The Z. montanum extract (20% w/w) granules was administered orally at doses of 0.3 – 1125 mg/kg/day to male and female Sprague Dawley rats (200 – 280 g) for 270 days. The toxicity effect was observed for 270 days and showed no clinical toxicity signs and no mortality, however, caused significant (p < 0.05) decrease in kidney and spleen weights in high dose female rats. The oral no-observed-adverse effect level (NOAEL) for Z. montanum extract (20% w/w) granules was 1125 mg/kg/day [ 52 ].

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.

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