Comparison study for the Phytochemical Constituents of two Curcuma species by GC-MS Technique

Introduction

Curcuma caesia Roxb. and Curcuma angustifolia Roxb. are two species belonging to the Curcuma genus, which is a part of the Zingiberaceae family. Curcuma angustifolia Roxb. is distributed in the Northeast, Madhya Pradesh, Uttar Pradesh, and Himachal Pradesh, while Curcuma caesia Roxb. is found in the Indian states of West Bengal, Orissa, Uttar Pradesh, Madhya Pradesh, Sikkim, and Chhattisgarh. In terms of importance and use, Curcuma angustifolia Roxb.is used in the production of arrowroot powder, and its rhizomes are used to make food. It is also used to treat diarrhoea, fever, and pain, and is a demulcent and blood-clotting agent.¹ Curcuma caesia Roxb.  is utilised as a folk medicine and has antiasthmatic, anticancer, antiallergy, and anti-inflammatory qualities. The rhizome has been used as a condiment and food preservative. ^2,3

Medicinal plants are appreciated for their therapeutic phytoconstituents, which may lead to the development of new medications. Because of the phytochemicals found in medicinal plants, as well as the pharmaceutical and cosmetic industries’ shift towards organic products, medicinal plant research is just as essential as traditional drug research.⁴ Phytoconstituents from plants must be extracted and measured in order to discover novel compounds or use them as a lead molecule in the production of more effective medicinal molecules.⁵ Extraction is critical in phytochemical processing for identifying and evaluating bioactive phytoconstituents from plant sources. Common extraction methods include decoction, hot continuous extraction, percolation, maceration, infusion, and others.⁶ Modern extraction methods, such as supercritical fluid extraction (SFE), microwave-assisted extraction (MAE), and ultrasound-assisted extraction (UAE), are constantly being developed to improve production while lowering costs.⁴ Heat extraction has the potential to destroy thermolabile natural substances. Ultrasound-Assisted Extraction (UAE) accelerates and cools the extraction process. Ultrasound cavitates and ruptures cell walls, accelerating the extraction of active plant components from the matrix and enhancing mass transfer.⁷

Solvent plays a vital part right through the extraction process and the type of solvent to be used is determined based on the nature of phytoconstituents to be extracted. For the extraction of the nonpolar secondary metabolites from plants, non-polar solvents are usually used. The extracts are subjected to GC-MS testing  to understand the phytoconstituents present and to uncover information about their mass and structures. ^8–11  .  The use of UAE in chloroform extracts of Curcuma caesia Roxb. rhizomes are reported in this study for the first time. There have been no reports of Curcuma angustifolia Roxb. rhizome extracts being investigated by the UAE. This study of the phytoconstituents of a UAE rhizome chloroform extract sheds insight on its therapeutic potential The information presented in this study will help reaffirm the usage of Curcuma caesia Roxb. and Curcuma angustifolia Roxb. rhizomes as reserves for medicinal phytoconstituents and can provide a pathway for developing herbal products based on the identification of the components and  understanding its nature from the present study..

Materials and Methods

Plant Collection

The Curcuma caesia Roxb. rhizome was gathered in March 2020 and 2021 from the ICAR-IISR (Indian Institute of Spices Research Kozhikode), located in Kerala. ICAR-IISR  authenticated and maintained the rhizome under accession number Acc. 292 (IC 349014). 

Curcuma angustifolia Roxb. rhizome was obtained from Jorhat, Assam, in the month of February every year between 2020-2021.Plant authentication with Accession No. was given by CSIR-National Botanical Research Institute (NBRI) Herbarium (LWG) 109910.

Chemicals

Analytical grade chloroform was used for the study. Whatman Filter Paper 41 and MDI 0.45 micron nylon syringe filters were used for the filtration of extracts.

Sample preparation

GC-MS Analysis

The rhizomes underwent a thorough scrubbing with water, followed by division into small pieces and subsequent drying for five days in shade. They were then powdered in a mixer grinder and sifted to obtain a fine powder. The powdered rhizomes were stored in covered jars and used for upcoming experiments.  . The rhizome powder and the solvent (chloroform) were taken in a ratio of 1:25 and sonicated for 30 minutes using an ultrasound sonicator water bath.¹³ The extraction was performed in a pulsed manner over 11 and 19 minutes, with 2-3 minute breaks in between. The temperature was maintained between 25 °C and 30 °C. Following sonication, ashless filter paper 41 from Whatman was used to sift the extracts, which were then filtered again through a nylon syringe filter of 0.45 micron before being introduced into the GC-MS instrument.

Phytochemical screening

The extracts filtered through Whatman Filter Paper 41 were utilized in the qualitative phytochemical screening studies.

Instrumentation

Sonicator

The Dakshin, 200H sonicator had a stainless steel tank with a capacity of 6.5 litres, an ultrasonic frequency of 36 ± 3 kilohertz (KHz), and an ultrasonic power of 200 watts. It required an electric supply of 230 volts A.C, 50 Hz, and could operate at a maximum temperature of 60 °C.

GC-MS

The GC-MS is a Shimadzu GCMS-QP2010 Plus model coupled with the GCMS-QP2010 Ultra Mass Spectrometer.

GC Condition

The analysis by GC-MS was carried out on a capillary column, Restek Rtx-5MS, having dimensions measuring 30 metres x 0.25 millimetres  ID, 0.25 micron with helium being the carrier gas and pressure as the flow control mode set at 83.5 kilopascals (kPa). The oven was programmed to heat at 80 °C during the first 5 minutes, then upstretched to 150 °C at a scale of 10 °C each minute and kept for 2 minutes, afterwards elevated to 220 °C at an amount of 10 °C for every minute and placed constant for 2 minutes, and ultimately enhanced to 290 °C at the same proportion of 10 °C every single minute and stationed for 5 minutes. The entire duration of the run was 35 minutes.¹⁴ The temperature of the injector port was 250 °C. The volume injected was 2 microlitres (µl) with a split ratio of 90.0.

MS Condition

The ion source and interface were heated to 220 °C and 260 °C, respectively. The detector gain was set at 1.03 kilovolts (kV). The solvent cut time was maintained at 2.0 minutes, and the scan start time and end time were 2.0 minutes and 34.0 minutes, respectively, with a start mass to charge (m/z) of 35.0 and an end m/z of 700.0. The scan speed was set at 2500. The data processing of mass spectra and chromatograms was performed using LabSolutions’ GCMS Solution Version 2.70 software. The NIST 11 library database was used for identifying the chemical components.

Results

A crucial tool in the bioactive component investigations, phytochemical screening is an easy and rapid process that provides a quick answer to the various types of phytochemicals in the extracts. This screening helps to gain awareness of the types of phytochemicals that are existent in the extracts.¹⁵ The outcomes of the phytochemical diagnosis of both extracts presented in Table 1 reveal the existence of steroids, terpenoids, phenolic compounds, and cardiac glycosides, but tannins, quinones, flavonoids, alkaloids, and amino acids are absent.

Table 1: Phytochemicals screening data for Curcuma caesia Roxb.and Curcuma angustifolia Roxb. chloroform extracts

Phytochemical Constituents	Test	Observation		Inference		References
		C. caesia	C. angustifolia	C. caesia	C. angustifolia
Terpenoids	extract + 1ml CHCl3 + few drops of concentrated. H2SO4	reddish brown colour interface appears	reddish brown colour interface appears	positive	positive	16
Steroids	extract + 5ml CHCl3 + 5ml concentrated. H2SO4	reddish top layer, sulphuric acid layer changed to yellowish	reddish top layer, sulphuric acid layer changed to yellowish	positive	positive
Tannins	extract + 1 ml of 5% FeCl3	Two layers observed. upper red and lower yellow	Two layers observed. upper red and lower yellow	negative	negative
Amino acid	extract + 3 drops of 5% lead acetate and heat the resulted solution	No blue/purple colour was observed	No blue/purple colour was observed	negative	negative
Flavonoids	extract + 2ml of 10% lead acetate	Two layers observed. and lower yellow and upper layer colourless	Two layers observed. and lower yellow and upper layer colourless	negative	negative
Alkaloids	extract + few drops of picric acid solution (in alcohol)	No yellow-coloured precipitate observed	No yellow-coloured precipitate observed	negative	negative
Phenolic compounds	extract + few drops of diluted Iodine solution	Transient reddish colour observed	Transient reddish colour observed	positive	positive	17
Quinone	extract + 4 drops of Isopropyl alcohol + 1ml of concentrated. H2SO4	Wine red colour observed	Wine red colour observed	positive	positive
Cardiac Glycosides	extract + 1ml glacial acetic acid + 1ml FeCl3 + 4 drops of concentrated. H2SO4	Slight greenish blue colour observed	Slight greenish blue colour observed	positive	positive

The assessment of phytoconstituents encompassed in the prepared extract from Curcuma caesia Roxb. and Curcuma angustifolia Roxb. rhizomes were carried out, and the results obtained were shown in Figures 1 and 2, respectively,  

Figure 1: GC-MS spectrum of Curcuma caesia Roxb. rhizome extract. Click here to View Figure

Figure 2: GC-MS spectrum of Curcuma angustifolia Roxb. rhizome extract Click here to View Figure

The GC_MS spectrum for Curcuma caesia Roxb. rhizome extract showed 26 peaks with different retention times, and peak areas with molecular weights for each identified compound, as shown in Table 2.

Table 2: Phytoconstituents in Curcuma caesia Roxb. rhizome chloroform extract

Peak No	Name	Retention Time (min)	Area %	Molecular weight	Molecular formula
1	Eucalyptol	3.558	2.15	154	C10H18O
2	Bicyclo[7.2.0]undec-4-ene, 4,11,11-trimethyl-8-methylene-,[1R-(1R*,4Z,9S*)]-	10.117	2.91	204	C15H24
3	(-)-Aristolene	10.815	6.50	204	C15H24
4	n-Tridecan-1-ol	11.143	1.73	200	C13H28O
5	Phenol, 2,4-bis(1,1-dimethylethyl)-	13.101	1.40	206	C14H22O
6	2H-3,9a-Methano-1-benzoxepin, octahydro-2,2,5a,9-tetramethyl-, [3R-(3.alpha.,5a.alpha.,9.alpha.,9a.alpha.)]-	14.004	1.10	222	C15H26O
7	1-Pentadecene	14.558	1.22	210	C15H30
8	2-Cyclohexen-1-one, 4-ethynyl-4-hydroxy-3,5,5-trimethyl-	14.912	24.42	178	C11H14O2
9	Bufa-20,22-dienolide, 3-hydroxy-15-oxo-, (3.beta.,5.beta.,14.alpha.)-	16.646	0.91	384	C24H32O4
10	1-Nonadecene	17.766	1.56	266	C19H38
11	Cyclohexane, 1,1-bis(5-methyl-2-furyl)-	19.317	2.20	244	C16H20O2
12	1,6-Dimethyl-9-(1-methylethylidene)-5,12-dioxatricyclo[9.1.0.0(4,6)]dodecan-8-one	19.742	1.84	250	C15H22O3
13	n-Hexadecanoic acid	19.913	0.74	256	C16H32O2
14	2,11-Dioxatetracyclo[4.3.1.1(3,10).0(6,9)]undec-4-ene, 3,7,7,10-tetramethyl-	20.067	2.16	206	C13H18O2
15	n-Tetracosanol-1	20.215	1.30	354	C24H50O
16	2,2,7,7-Tetramethyltricyclo[6.2.1.0(1,6)]undec-4-en-3-one	20.384	15.59	218	C15H22O
17	1,2-Dimethyl-5-nitroadamantane	20.722	3.05	209	C12H19NO2
18	Phenol, 3-ethyl-, acetate	20.860	1.48	164	C10H12O2
19	2-(1-(Beta-d-glucopyranosyloxy)-1-methylethyl)-2,3-dihydro-7-oxo-7H-furo(3,2-g)chromene, (R)-	20.996	5.72	408	C20H24O9
20	Pregn-4-ene-1,20-dione, 12-hydroxy-16,17-dimethyl-	21.289	9.04	358	C23H34O3
21	2-Propenal, 3-(2,6,6-trimethyl-1-cyclohexen-1-yl)-	21.646	8.93	178	C12H18O
22	Cyclopropa[c,d]pentalene-1,3-dione, hexahydro-4-(2-methyl-2-propenyl)-2,2,4-trimethyl-	22.638	1.23	232	C15H20O2
23	Cyclohex-2-enone, 3-(N’,N’-dimethylhydrazino)-5-(3-methoxyphenyl)-	23.223	1.27	260	C15H20N2O2
24	1-Nonadecene	25.131	0.62	266	C19H38
25	Purine-2,6-dione, 8-(3-ethoxypropylamino)-1,3-dimethyl-3,9-dihydro-	27.847	0.42	281	C12H19N5O3
26	1,10-Diazacyclooctadecane	28.186	0.51	254	C16H34N2

 

The chromatographic sketching showed the presence of phenolic compounds, terpenoids, steroid and aromatic compounds. 2-Cyclohexen-1-one, 4-ethynyl-4-hydroxy-3,5,5-trimethyl- referred as 2CEHT was the component with the highest area concentration with 24.42%, followed by 2,2,7,7-Tetramethyltricyclo[6.2.1.0(1,6)]undec-4-en-3-one with 15.59%, Pregn-4-ene-1,20-dione, 12-hydroxy-16,17-dimethyl- with 9.04%, 2-Propenal, 3-(2,6,6-trimethyl-1-cyclohexen-1-yl)- with 8.93%, and (-)-Aristolene with 6.50% were the other major components detected. Eleven components had an area concentration greater than 2%.

The results also indicate 32 peaks in the GC-MS spectra for Curcuma angustifolia Roxb. rhizome extract with different retention times, and peak areas, with molecular weights for each identified compound as shown in Table 3.

Table 3: Phytoconstituents in Curcuma angustifolia Roxb. rhizome chloroform extract

Peak No	Name	Retention Time (min)	Area %	Molecular  weight	Molecular formula
1	(+)-2-Bornanone	6.411	6.62	152	C10H16O
2	Isoborneol	6.729	4.67	154	C10H18O
3	2-Cyclohexen-1-one, 4-ethynyl-4-hydroxy-3,5,5-trimethyl-	14.916	20.55	178	C11H14O2
4	2-Naphthalenemethanol, decahydro-.alpha.,.alpha.,4a-trimethyl-8-methylene-, [2R-(2.alpha.,4a.alpha.,8a.beta.)]-	15.743	2.98	222	C15H26O
5	1-Naphthalenol, decahydro-1,4a-dimethyl-7-(1-methylethylidene)-, [1R-(1.alpha.,4a.beta.,8a.alpha.)]-	15.819	1.49	222	C15H26O
6	3,7-Cyclodecadien-1-one, 3,7-dimethyl-10-(1-methylethylidene)-, (E,E)-	16.470	1.34	218	C15H22O
7	dl-Phenylephrine	16.835	0.52	167	C9H13NO2
8	Tricyclo[4.3.1.1(3,8)]undecane-1-carboxylic acid	16.915	1.75	194	C12H18O2
9	Acetic acid, trifluoro-, octahydro-4-hydroxy-1,5-methano-1H-inden-1-yl ester (1.alpha.,3a.beta.,4.beta.,5.beta.,7a.beta.)-	17.515	1.41	264	C12H15F3O3
10	Propanoic acid, 2-[(1-cyclohexylethyl) carbamoyl]-, ethyl ester	17.590	0.47	255	C14H25NO3
11	(-)-Spathulenol	17.644	1.78	220	C15H24O
12	2-Methyl-6,7-dihydro-5H-benzofuran-4-one	17.796	0.31	150	C9H10O2
13	1-(3,5-Dimethyl-1-adamantanoyl) semicarbazide	17.834	0.11	265	C14H23N3O2
14	2-Propen-1-amine, N, N-di-2-propenyl-	18.042	0.36	137	C9H15N
15	1-Indolinecarboxaldehyde, 2-hydroxy-5-methoxy-	18.181	0.37	193	C10H11NO3
16	2,11-Dioxatetracyclo [4.3.1.1(3,10).0(6,9)] undec-4-ene, 3,7,7,10-tetramethyl	19.320	2.84	206	C13H18O2
17	1,6-Dimethyl-9-(1-methylethylidene)-5,12-dioxatricyclo [9.1.0.0(4,6)] dodecan-8-one	19.747	2.36	250	C15H22O3
18	Columbin	19.820	0.41	358	C20H22O6
19	n-Hexadecanoic acid	19.924	3.69	256	C16H32O
20	2,11-Dioxatetracyclo [4.3.1.1(3,10).0(6,9)]undec-4-ene, 3,7,7,10-tetramethyl	20.068	3.65	206	C13H18O2
21	6-(1-Hydroxymethylvinyl)-4,8a-dimethyl-3,5,6,7,8,8a-hexahydro-1H-naphthalen-2-one	20.209	1.77	234	C15H22O2
22	Androst-5-en-7-one, 3-(acetyloxy)-4,4-dimethyl-, (3.beta.)-	20.563	0.85	358	C23H34O3
23	1,2-Dimethyl-5-nitroadamantane	20.724	4.75	209	C12H19NO2
24	4-Isopropyl-3,4-dimethylcyclohexa-2,5-dienone	20.860	1.65	164	C11H16O
25	2-(1-(Beta-d-glucopyranosyloxy)-1-methylethyl)-2,3-dihydro-7-oxo-7H-furo(3,2-g)chromene, (R)-	20.999	6.21	408	C20H24O9
26	Spiro[2,4,5,6,7,7a-hexahydro-2-oxo-4,4,7a-trimethylbenzofuran]-7,2′-(oxirane)	21.294	7.52	208	C12H16O3
27	Diethylmalonic acid, 2-methoxyethyl tetradecyl ester	21.451	2.09	414	C24H46O5
28	4H-1,3,2-Dioxaborin, 4,6-diethenyl-2-ethyl-4-methyl-	21.647	10.00	178	C10H15BO2
29	Oxacyclopentadecan-2-one, 15-methyl-	21.892	0.87	240	C15H28O2
30	Diisooctyl maleate	22.005	1.71	340	C20H36O4
31	Octadecanoic acid	22.169	2.51	284	C18H36O2
32	Isoxazole, 5-chloro-4-(2-phenylethyl)-	25.859	2.37	207	C11H10ClNO

 

The components class detected comprises of sesquiterpenoid alcohols, terpenoids, fatty acids and phenolic compounds. 2CEHT with 20.55%, was the component with the highest area concentration. 4H-1,3,2- Dioxaborin, 4,6-diethenyl-2-ethyl-4-methyl- with 10.00%, Spiro[2,4,5,6,7,7a-hexahydro-2-  oxo-4,4,7a-trimethylbenzofuran]-7,2′-(oxirane) with 7.52 %, (+)-2-Bornanone with 6.62, 2- (1-(beta-d-glucopyranosyloxy)-1-methylethyl)-2,3- dihydro-7-oxo-7H-furo(3,2-g) chromene,  (R)- denoted as 2BGDFC with 6.21, 1,2-dimethyl-5- nitroadamantane with 4.75%, and  isoborneol with 4.67% were some of the other major components detected. Fifteen components had an area concentration greater than 2%.          

Discussion

GC-MS method conditions

By carefully establishing the analysis method conditions, the extracts underwent GC-MS screening in order to achieve uniform peak responses, optimal peak separation, and peak resolution. The detectable peaks were identified by name, mass, and structure. A nonpolar column, Restek Rtx-5MS, with an optimum length of 30 m, a moderate ID of 0.25 mm, and a thin film thickness of 0.25 micron, was used for the study. Helium was used as the carrier gas due to its inertness, safety, and its ability to provide good separations.

A gradient temperature programme was finalised for faster elution of components from the column. A constant pressure mode was used for reduced consumption of carrier gas.¹⁸ The initial column oven temperature was 100 degrees lower than the injection surface temperature, as this difference encouraged the analyte concentration to reach the column head at the earliest.¹⁹ Direct injection was used for injecting the sample, as the heat at the injection interface vaporised the sample mixture before it entered the column.

To minimise the quantity of plant extract entering the column, a split ratio of 90 was used, resulting in narrow and sharp peaks. Ion source temperatures of 220°C were vital for EI ionisation.

GC-MS assessment findings

Six components were found to be common in the GC-MS data of both curcuma species. The area percentages of these phytoconstituents were equated graphically for better understanding in Figure 3.

Figure 3: Graphical comparison of area percentages of common phytoconstituents found in Curcuma caesia Roxb. and Curcuma angustifolia Roxb. chloroform extracts. Click here to View Figure

From the graphical representation, it is evident that the area concentration of 2CEHT is present in higher amounts as compared to other phytoconstituents in both species. This phytoconstituent is reported to be present in the essential oil of a plant species that demonstrates anticancer activity.²⁰

The data from GC-MS examination of the rhizome chloroform extract of Curcuma caesia Roxb. prepared by UAE from this investigation were judged with the GC-MS statistics of the chloroform extract generated by Soxhlet extraction by Atom et al.¹² In contrast to the twenty-six components discovered in the GC-MS study of the of Curcuma caesia Roxb.rhizome chloroform extract prepared by the UAE, twenty components were detected in the chloroform extract of Curcuma caesia Roxb. rhizomes prepared by Soxhlet extraction. Phenol, 2,4-bis(1,1-dimethylethyl)-, a phenolic component, was found common in both studies.

The ability of plants or herbs to treat disease depends on the phytochemical makeup of those substances, which displays a variety of intriguing and unique biological functions. It has been found that the various phytochemicals identified in this study have a wide range of biologic functions.²¹ Table 4 lists the bioactivities reported for the phytoconstituents detected in the GC-MS evaluation of Curcuma caesia Roxb. and Curcuma angustifolia Roxb. rhizomes extracted in chloroform by UAE.

Table 4: Biological activities reported for some phytoconstituents detected in GC-MS analysis

Curcuma caesia Roxb. rhizomes
Peak No	Retention Time	Name	Area %	Activity	References
1	3.558	Eucalyptol	2.15	anti-inflammatory	22
3	10.815	(-)-Aristolene	6.50	Insecticidal	23
5	13.101	Phenol, 2,4-bis(1,1-dimethylethyl)-	1.40	larvicidal, repellent, acaricidal	24
8	14.912	2CEHT	24.42	anticancer	20
13	19.913	n-Hexadecanoic acid	0.74	antibacterial	25
15	20.215	n-Tetracosanol-1	1.30	antimutagenic	26
19	20.996	2BGDFC	5.72	Improving working memory dysfunction	27
Curcuma angustifolia Roxb. rhizomes
1	6.411	(+)-2-Bornanone	6.62	Antineoplastic, pain reliever, microbicidal, Inflammation reducer, antimycotic	28
2	6.729	Isoborneol	4.67	use against atherosclerotic disease	29
3	14.916	2CEHT	20.55	anticancer	20
11	17.644	(-)-Spathulenol	1.78	anti-inflammatory, antimicrobial, anti-proliferative, antioxidant, antifungal, antibacterial	30–32
19	19.924	n-Hexadecanoic acid	3.69	antibacterial	25
25	20.999	2BGDFC	6.21	Improving working memory dysfunction	27

As certain phytoconstituents detected in Curcuma caesia Roxb. and Curcuma angustifolia Roxb. rhizomes demonstrate biological activity, the chloroform extracts of both these species can be standardised further and used as herbal medications for reported ailments. These phytoconstituents can be used as marker compounds and quality control tools in the standardisation of plant extracts. The data presented in this study gives scientists the chance to investigate phytoconstituents that have not yet been linked to any biological activity.

Additionally, UAE had previously been utilised to unearth metabolites, ecological pigments, and bioactive compounds from other curcuma species.^33–40 The detection of bioactive phytoconstituents in the GC-MS examination of rhizome chloroform extracts of Curcuma caesia Roxb. and Curcuma angustifolia Roxb. extracted by UAE defines a straightforward approach for the extraction of phytoconstituents from different plant parts with minimal processing time.

Conclusions

The study assessed chloroform ultrasonic-assisted extracts of Curcuma caesia Roxb. and Curcuma angustifolia Roxb. rhizomes by GC-MS analysis and phytochemical screening. Results showed significant phytoconstituent extraction, highlighting the need for recurring usage of UAE in plant standardisation studies. The extracts exhibited potential therapeutic use that can be converted to prospective novel medications by performing further studies for the isolation and separation of the bioactive components found in the study. Overall, Curcuma caesia Roxb. and Curcuma angustifolia Roxb. could be crucial sources of medicine in contemporary treatment.

Acknowledgements

The authors gratefully recognise and convey their thanks for the support given by Dr. Siddhartha. P. Saikia, Principal Scientist & Head, Agrotechnology and Rural Development Division, CSIR-NEIST, Jorhat, Assam, in providing Curcuma angustifolia rhizomes.

The authors appreciatively acknowledge and express thanks for the help rendered by Dr. D. Prasath, Principal Scientist, ICAR-IISR, Kozhikode, Kerala, in sharing the Curcuma caesia rhizomes.

Conflict of Interest

The authors state that they have no competing interests to declare.

Funding Sources

There is no funding sources

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