Temu hitam Rhizome
Curcuma aeruginosa Roxb.
Zingiberaceae
Figure 1 : Curcuma aeruginosa. (a) Whole plant, (b) leaf and (c-d) rhizome. (Photos courtesy of Mohd Fauzi Sayuti, UKM, 2015)
DEFINITION
Temu hitam rhizome consists of the powder of dried rhizomes of Curcuma aeruginosa Roxb. (Zingiberaceae).
SYNONYM
Information and data have not been established [ 1 ].
VERNACULAR NAMES
Pink and blue ginger (English); Temu hitam (Malay); Ngo suk, ezhu (Chinese); Karimanjal (Indian) [ 2 , 3 , 4 , 5 , 6 ].
CHARACTER
IDENTIFICATION
Plant Morphology
C. aeruginosa is a rhizomatous herb 70–100 cm tall; pseudostem 30–35 cm tall and green sheath. Rhizome is 5–6 x 9–10 cm, blue in the centre, merging towards grey, the blue colour is highly variable with age of the rhizome, strongly aromatic; sessile tubers branched, condensed; roots fleshy; root tubers many, ovate-oblong, creamy; short and smooth fracture. Leaves are distichous, 30–40 × 10–12 cm; lamina is oblong-lanceolate with acute apex, acuminate base, glabrous purple or reddish-brown patch along the sides of the distal half of the midrib on the adaxial and fading at maturity, green groove of the midrib; petiole is as long as lamina. Inflorescence is lateral, 25–30 cm long; peduncle is 12–18 cm; spike is 12–15 × 5 cm; coma bracts are large, pink to violet and lower ones are streaked green; fertile bracts are 18–20, green with a pink tip, 4.5–5 × 4.4–5 cm, lower half is fused, tip is rounded; bracteoles 3.5 × 2.5 cm, white with a median light green patch. Flowers are 4.5–5 cm, in cincinnus of 8–10 in a bract, equal to or slightly shorter than the bracts; calyx is 1 cm, truncate, 3-lobed at apex, split on one side; corolla tube is 3–3.3 cm long, pink, lobes unequal; dorsal lobes are 1.5 × 1.2 cm, concave, hooded; lateral lobes are 1.5 × 1 cm, tip rounded, pink; labellum is 1.5–1.7 × 1.8 cm tip emarginated, yellow with a deep yellow median band; lateral staminodes are 1.5 × 1 cm and yellow; anther is 7 mm, without crest, spurred at base, spurs are 3 mm long and divergent; epigynous glands are two, 5 mm long, linear and yellowish green; ovary is 5 mm, trilocular, with many ovules; style is long, filiform; stigma bilipped, slightly exerted above the anther lobes [ 7 ].
Microscopy
Powdered material consists of rectangular parenchyma cells; simple starch granules that are oval-ellipsoidal and sometimes polyglonal in shape with hilum circular or 2–5 rayed cleft; long unicellular trichomes; annular and reticulate vessels; bundle of lignified fibres and prism calcium oxalate crystals [ 7 ].
Figure 2 : Microscopic characters of Curcuma aeruginosa dried rhizome powder of 0.106 mm size. (a) Parenchyma cells (magnification 100×); (b-c) starch granules (magnification 400×); (d) unicellular trichome (magnification 100×); (e) reticulate vessel (R) and annular vessel (A) (magnification 100×); (f) lignified fibres (magnification 100×); (g-h) prism calcium oxalate crystals (magnification 400×). [Scale bars: a, d-f = 100 µm; b-c, g-h= 20 µm]
Colour Tests
Observed colour of solution after treatment with the following reagent:
| KOH (5%) | Brown |
Thin Layer Chromatography (TLC)
Figure 3 : TLC profiles of curcumin (S), methanol extract of Curcuma aeruginosa dried rhizome powder (L) observed under (a) visible light after derivatisation with four prominent bands.
| Test Solutions | Weigh about 5.0 g of C. aeruginosa dried rhizome powder of 0.106 mm size in a 100 mL screw cap conical flask. Add 50 mL of methanol and sonicate for 30 minutes (without temperature setting). Filter the mixture with filter paper and use the filtrate as test solution. |
|
Standard solution (for system suitability test) |
Dissolve curcumin standard [CAS no. 458-37-7] in methanol to produce a standard concentration 0.3 mg/mL solution. |
| Stationary Phase | HPTLC Silica gel 60 F254, 10 x 10 cm |
| Mobile phase | Dichloromethane-methanol, 25:1 (v/v) |
| Application |
|
| Development distance | 8 cm |
| Drying | Air drying |
| Detection |
|
High Performance Liquid Chromatography (HPLC)
| Test solution | Weigh about 5.0 g of C. aeruginosa 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 minutes 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 methanol to produce 0.3 mg/mL solution. |
| Chromatographic system |
Detector: UV 430 nm Column: C18 (5 μm, 4.6 mm x 250 mm) (Zorbax Eclipse XDB –C18 if necessary) Column oven temperature: 25oC Flow rate: 0.4 mL/min Injection volume: 10 µL |
| Mobile phase (Isocratic mode) | Acetonitrile : 0.1% orthophosphoric acid in water (55:44 v/v) |
| Run time | 30 min |
| System suitability requirement |
Perform at least five replicate injections of the standard solution (0.3 mg/mL). The requirements of the system suitability parameters are as follow:
|
| Acceptance criteria |
|
(a)
(b)
Figure 4 : Whole HPLC chromatogram of (a) demethoxycurcumin in curcumin standard solution (0.3 mg/mL) at tr= 16.322 min and (b) ethanol extract of Curcuma aeruginosa dried rhizome powder showing peak corresponding to demethoxycurcumin in curcumin standard solution at tr= 16.300 min.
(a)
(b)
Figure 5 : HPLC chromatogram highlighting the elution region of demethoxycurcumin in (a) curcumin standard solution (0.3 mg/mL) at tr= 16.322 min and (b) ethanol extract of Curcuma aeruginosa dried rhizome powder showing peak corresponding to demethoxycurcumin in curcumin standard solution at tr= 16.300 min.
Table 1: The Relative Retention Time (RRT) of the characteristic peaks for all of the standards used
| Standard | RRT (min) |
| bisdemethoxycurcumin | 14.772 |
| demethoxycurcumin (as reference) | 16.322 |
| curcumin | 18.103 |
Note: The RRTs listed above serve only as guidance
PURITY TESTS
The purity tests are based on C. aeruginosa 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 5% |
| Loss on Drying |
| Not more than 12% |
| Extractive Values | |
| Water-soluble extracts | |
| Hot method | Not less than 17% |
| Cold method | Not less than 8% |
| Ethanol-soluble extracts | |
| Hot method | Not less than 5% |
| Cold method | Not less than 3% |
SAFETY TESTS
The safety tests are based on C. aeruginosa 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
Methanol extract was found to contain guaine sesquiterpene lactones (e.g. zedoalactone A, zedoalactone B, zedoarondiol) [ 8 ].
Chloroform extract was found to contain sesquiterpene (e.g. isocurcumenol) [ 9 ].
Petroleum ether extract was found to contain sesquiterpenes (e.g. zedoarol, curcumenol) [ 9 ].
Hexane extract was found to contain sesquiterpenes (e.g. germacrone, zederone, dehydrocurdione, curcumenol, zedoarondiol, isocurcumenol) [ 3 ].
The essential oils of C. aeruginosa rhizomes has been found to contain monoterpenoids (e.g. isoborneol, α-pinene, sabinene, α-terpene, camphor, borneol, tumerone, ar-tumerone, zerumbone, terpinen-4-ol, furanodienone, camphene, α-terpineol, β-pinene, carvone, curcumenol, myrcene, isocurcumenol, 1,8-cineole, α-selinene, β-selinene, linalool, mycrene, limonene, β-bisabolene, δ-cadinene, β-sesquiphellandrene, curzerene, T-cadinol, T-muurolol, α-cadinol, epicurzerenone, 3-carene, 2-thujene, epi-bicyclosesquiphellandrene), caryophyllane-type sesquiterpenoids (e.g. caryophyllene, β-caryophyllene, caryophyllene oxide, β-caryophyllene oxide), germacrane-type sesquiterpenoids (e.g. germacrone, germacrene D, germacrene B, germacrone, 8,9-β-dimethyl-4α,9β-dihydrodibenzo[b,d]furan-3(4h)-one, isofuranogermacrene), elemane-type sesquiterpenoids (e.g. β-elemene, α-elemene, δ-elemene), sesquiterpenoids (e.g. humulene, β-farnesene, (Z)-α-farnesene, (Z)-β-farnesene, β-cubebene, ar-curcumene) and other substances (e.g. eucalyptol; L-camphor; eudesma-4(14),11-diene; α-bulnesene; cycloisolongifolene, 8,9-dehydro-9-formyl; alloaromadendrene oxide-(2); dihydrocostunolide; velleral; zedoarol; 2-nonanone; 2-undecanone; myrtenol; trans-pinocarveol; acetophenone; propiolic acid, 3-(1-hydroxy)-2-isopropyl-1,5-methylcyclohexyl); 4-oxo–β-isodamascol; methenolone; labd-13-en-15-oic acid,8,12-epoxy-12-hydroxy-γ-lactone; cholesta-22,24-dien-5-ol-4,4-dimethyl-; β-sitosterol; heptan-2-ol; tricyclene; ρ-cymene; nonan-2-ol; linalol; ρ-cymene-4-ol; undecan-2-one; undecan-2-ol; α-guajene; selina-3,7(11)-diene, β-eudesmol, humuladienone, curdione, curcumanolides A and B; dehydrocurdione; curcumenone; oxalic acid, bis(trimethylsilyl) ester; malonic acid bis(trimethylsilyl) ester; butanoic acid, 4-[(trimethylsilyl)oxy]-, trimethylsilyl ester; L-alanine, N-octanyl-ethyl ester; butanedioic acid, [(trimethylsilyl) oxy]-, bis(trimethylsilyl) ester; citric acid, ethyl ester, tri-trimethylsilyl; isocitric acid (trimethylsilyl); D-fructose-1,3,4,5,6-pentakis-O-(trimethylsilyl)-O-methyloxime; D-glucose-2,3,4,5,6-pentakis-O-(trimethylsilyl)-O-methyloxime; hexadecanoic acid, trimethylsilyl ester; myo-inositol,1,2,3,4,5,6-hexakis-O-(trimethylsilyl); 4,4-dimethyl-N-(2-phenylethyl)-5α-androst-2-en-17-amine; stearic acid, trimethylsilyl ester; tetracosane; 17-hydroxy-3,20-dioxopregna-1,4,9(11)-trien-21-yl acetate; triacontane; tetratriacontane; α-D-glucopyranoside-1,3,4,6-tetrakis-O-(trimethylsilyl)-β-D-fructofuranosyl-2,3,4,6-tetrakis-O-(trimethylsilyl); 3-methyl cyclopentane-1-yl-trimethylsilyl ether; thiosalicylic acid O, S-di-trimethylsilyl-; butane-1,3-diol; 1-methylene-3-methyl-,bis(trimethylsilyl)ether; glycine, N-(trimethylsilyl)-, trimethylsilyl ester; borneol-trimethylsilyl ether, phenylethanolamine; tris (trimethylsilyl) phosphate; (2,6-ditert-butylphenoxy)(trimethyl) silane; formic acid, 2-bro,omethyl-4, 4-dimethyl-3-(3-oxobut-1-enyl)cyclohex-2-enyl ester; germacra-1(10),4-diene-12-oic acid 6-α-hydroxy gamma lactone; propiolic acid, 3-(1-hydroxy-2-isopropyl-5-methylcyclohexyl); androst-4-en-17-one,3,16, bis(trimethylsilyl) oxy))-3α; 2-isopropenyl-2,3-dihydro-7H-furo (3,2-g) chromen-7-one; pregna-1,4,16-triene-3,20-dione,11,22-diacetoxy-; heptadecanoic acid, trimethylsilyl ester; linoleic acid, trimethylsilyl ester; oleic acid, trimethylsilyl ester; 4α-methylandrostane-2,3-diol-17-dione; anthiaergostan-5,7,9,22-tetraen-14-ol-15-one; 19-norpregn-4-en-20-yn-3-one, 17-(trimethylsilyl)oxy; androst-5-en-17-one,3,16-bis[(trimethylsilyl)oxy)],(O-methyloxime,(3β, 16α)) [ 1 , 7 , 10 , 11 , 12 , 13 , 14 , 15 ].
MEDICINAL USES
Uses described in folk medicine, not supported by experimental or clinical data
Traditionally used for cough and shortness of breath. It is also used externally and pounded with coconut oil for scurf [ 2 ].
Biological and pharmacological activities supported by experimental data
Cytotoxicity activity
Germacrone isolated from C. aeruginosa dried powdered rhizomes (10 µg/mL) showed significant (p < 0.05) decrease in cell viability on human prostate cancer cells (LNCaP cell) (70%) compared to vehicle control group (100%) [ 3 ].
Anti-androgenic activity
Hexane extract of C. aeruginosa dried powdered rhizomes and germacrone isolated from the extract inhibited the conversion of testosterone to dihydrotestosterone (DHT) with IC50 values of 0.22 ± 0.03 and 0.42 ± 0.05 µg/mL, respectively, compared to the positive control, ethinylestradiol (0.26 ± 0.02 µg/mL) using rat liver enzyme test [ 3 ].
Germacrone isolated from C. aeruginosa dried powdered rhizomes (3, 30 and 100 µg) was applied to flank glands of androgen-stimulated growth male Syrian golden hamster for 4 weeks. The compound showed significant (p < 0.05) increase in change of flank gland size (1.6 ± 2.7 – 1.9 ± 0.8 mm) compared to testosterone treated group (9.0 ± 1.8 mm) [ 3 ].
Anti-thrombocytopenia activity
Ethanol extract of C. aeruginosa dried rhizomes (250 and 500 mg/kg body weight) was administered orally to heparin‐induced thrombocytopenia male Wistar rats (150-250 g) for seven days. The extract increased the number of thrombocytes (250 mg/kg: 24.72%; 500 mg/kg: 26.95%), erythrocytes (250 mg/kg: 4.52%; 500 mg/kg: 9.53%) and hematocrits level (250 mg/kg: 2.28%; 500 mg/kg: 8.19%) [ 17 ].
Antinociceptive effect
Chloroform extract of C. aeruginosa dried rhizomes (400 mg/kg) administered orally to male Swiss mice (30 – 38 g) 30 min before induction of abdominal constriction using acetic acid significantly (p < 0.05) decreased the number of writhing by 10.7 ± 3.8 counts/20 min compared to positive control, aspirin (200 mg/kg) (9.6 ± 2.6 counts/20 min) [ 18 ].
Chloroform extract of C. aeruginosa dried rhizomes (400 mg/kg) administered to male Wistar rats (150 – 220 g) 30 min after induction of pain using formalin significantly (p < 0.05) decreased the licking activity in the late phase by 7.41 ± 4.24 sec compared to positive control, aspirin (200 mg/kg) (9.89 ± 16.83 sec) [ 18 ].
Clinical studies
Information and data have not been established.
SAFETY INFORMATION
Preclinical studies (Toxicology studies)
Hepatic necrosis
Chloroform extract of C. aeruginosa rhizome (0.004 g/kg and 0.06 g/kg BW) was administered orally to male Balb C mice (two-month old; 25 – 30 g) once a day for 10 days. On day 11, the mice were sacrificed and the livers were analysed. The extract showed significant (p < 0.05) result in mean percentage of hepatocyte apoptosis (0.004 g/kg BW: 1.49 ± 0.22%; 0.06 g/kg BW: 1.33 ± 0.24%) compared to control group 0.30 ± 0.13% [19 ].
Acute toxicity
Oral single dose acute toxicity study on female Sprague Dawley rats ( aged between 8 and 12 weeks old) using aqueous extract of C. aeruginosa rhizomes showed no toxic effect on the parameters observed, including behaviors, 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 more than 2,000 mg/kg body weight [ 20 ].
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
- The Plant List 2016. [Internet] Curcuma aeruginosa Roxb; 2013 [cited on 24 May 2016]. Available from: http://www.theplantlist.org/tpl1.1/record/kew-235184.
- Burkill IH. A dictionary of the economical products of Malay Peninsula. Volume 1. Kuala Lumpur: Ministry of Agriculture & Cooperative. 1966; pg. 716.
- Suphrom N, Pumthong G, Khorana N, Waranuch N, Limpeanchob N, Ingkaninan K. Anti-androgenic effect of sesquiterpenes isolated from the rhizome of Curcuma aeruginosa Roxb. Fitoterapia. 2012;83:864-871.
- Globinmed. [Internet] Curcuma aeruginosa Roxb. [cited on 25 May 2016]. Available from: http://www.globinmed.com/index.php?option=com_content&view=article&id=104732;curcuma-aerugenosa-1&catid=199&Itemid=139.
- Lim TK. Edible Medicinal an Non Medicinal Plants. Volume 12. Springer International Publishing Switzerland. 2016.
- Zhanwen Liu. Essentials of Chinese Medicine. Springer Science & Business Media. 2011:241.
- Srivastava S, Chitranshi N, Srivastava S, Dan M, Rawat AKS, Pushpangadan. Pharmacognostic evaluation of Curcuma aeruginosa Roxb. 2006;12(3):162-165.
- Takano I, Yasuda I, Takeya K, Itokawa H. Guaiane sesquiterpene lactones from Curcuma aeruginosa. Phytochemistry. 1995;40(4):1197-1200.
- Mohd Aspollah HS, Suhaila MS, Nordin HL, Mawardi R, Radzali M, Umi Kalsom Y, Sugeng R. Chemical constituents and bioactivity of Curcuma aeruginosa Roxb. Natural Product Sciences. 2007;13(3):175-179.
- Tg Kamazeri TSA, Abd Samah O, Taher M, Susanti D, Qaralleh H. Antimicrobial activity and essential oils if Curcuma aeruginosa, Curcuma mangga, and Zingiber cassumunar from Malaysia. Asian Pacific Journal of Tropical Medicine. 2012:202-209.
- Sirat H, Jamil S, Hussain J. Essential oil of Curcuma aeruginosa Roxb. from Malaysia. Journal of Essential Oil Research. 1998;10(4):453-458.
- Jarikasem S, Thubthimthed S, Chawananoraseth K, Suntorntanasat T. Essential oils from three Curcuma species collected in Thailand. Acta Hocticulturae. 2005;37-40.
- Simoh S, Zainal A. Chemical profiling of Curcuma aeruginosa Roxb. rhizome using different techniques of solvent extraction. Asian Pacific Journal of Tropical Biomedicine. 2015;5(5):412-417.
- Aromdee C, Polrat S, Wangsomnuek P. Constituents of the rhizome of Curcuma aeruginosa and its DNA fingerprint. International Journal of Phytomedicine. 2011;31:98-203.
- Zwaving JH, Bos R. Analysis of the essential oil of five Curcuma species. Flavour and Fragrance Journal. 1992;7:19-22.
- Ibrahim J, Abu Said A, Nor Azah MA, Abdul Rashih A, Halijah I. Chemical composition of the rhizome oils of four Curcuma species from Malaysia. Journal of Essential Oil Research. 1999; 11(6):719-723.
- Moektiwardoyo MW, Tjitraresmi A, Susilawati Y, Iskandar Y, Halimah E, Zahryanti D. The potential of dewa leaves (Gynura pseudochina (L) DC) and temu ireng rhizomes (Curcuma aeruginosa Roxb.) as medicinal herbs for dengue fever treatment. Procedia Chemistry. 2014;13:134-141.
- Reanmongkol W, Subhadhirasakul S, Khaisombat N, Fuengnawakit P, Jantasila S, Khamjun A. Investigation the antinociceptive, antipyretic and anti-inflammatory activities of Curcuma aeruginosa Roxb. Extracts in experimental animals. Songklanakarin Journal of Science and Technology. 2006;28(5):999-1008.
- Hestianah EP, Kusumawati I, Suwanti LT, Subekti S. Toxic compounds of Curcuma aeruginosa causes necrosis of mice hepatocytes. Universa Medicina. 2014;33(2):118-125.
- Norazlina Z, Emylyn M, Siau TC, Izwah H, Wan Abdul Hakim WL, Wan Mohammad Adham Afiq WZ, Teh BP, Hussin M. Acute oral toxicity study of selected Malaysian medicinal herbs on Sprague Dawley rats. Institute for Medical Research, Ministry of Health; 2015. Report no.: HMRC 11-045/01/CA/RH/C.
