A Comprehensive Review on Pharmacologically Active Phyto-Constituents from Hedychium species

In this review, we describe and discuss the phytoconstituents present in Hedychium species and emphasize their potential as drug candidates. Though they are widely validated in vitro and in vivo models, to date, no efforts have been made to compile in a single review all the pharmacologically active phytoconstituents from Hedychium species, and their pharmacological and toxicity profile. In this study, we present a reinvestigation of the chemical constituents present in Hedychium species obtained from the essential oil and solvent extraction of the flowers, leaves and rhizomes under consideration. Key databases such as PubMed, Science Direct, Scopus, and Google Scholar amongst others were probed for a systematic search using keywords to retrieve relevant publications on this plant. An exhaustive electronic survey of the related literature on Hedychium species resulted in around 200 articles. Articles published between the years 1975–2021 were included. The studies conducted on either crude extracts, solvent fractions or isolated pure compounds from Hedychium species reported with a varied range of biological effects such as anti-inflammatory, analgesic, antidiabetic, potentially anti-asthmatic, and cytotoxic, among other related activities of the chemical constituents present in its essential oil and solvent extract deployed in this review. Traditional and herbal medication around the world that uses different parts of Hedychium species were considered for anti-inflammatory, skincare, analgesic, anti-asthmatic, anti-diabetic, antidotal uses, among others. These uses support the idea that chemical constituents obtained from solvent extraction may also exert the same action individually or in a synergistic manner. The review concluded that there is scope for computation and biological study to find out possible new targets for strengthening the potency and selectivity of the relevant compounds, and to find a commercial method for extraction of active pharmaceutical ingredients.


Introduction
The Himalayas Mountain range is an abundant source of medicinally active plants, herbs, shrubs etc. There are many naturally growing plant species. One of the plant genera found in Himalaya region is Hedychium. We consider Hedychium species in this study. Hedychium species (spiked ginger lily) [1] are annual-perennial, rhizomatous, erect flowering plants. They are abundant with aromatic and medicinally active compounds. The herb belongs to the family Zingiberaceae. About 100 species of Hedychium are found Hedychium species (spiked ginger lily) [1] are annual-perennial, rhizomatous, erect flowering plants. They are abundant with aromatic and medicinally active compounds. The herb belongs to the family Zingiberaceae. About 100 species of Hedychiumare found worldwide [2], and the most-explored species are Hedychium spicatum [3], Hedychium coronarium, Hedychium coccineum, Hedychium flavescens, Hedychium gardnerianum [4] etc. They are situated in the subtropical area of Himalayan region, India, and widely found in China, Myanmar, Nepal, and Thailand, Madagascar (Africa), hot tropical regions of Asia, Indo-China, Malasia, Indonesia etc. [5]. Traditional applications and previous investigations carried out by many researchers on Hedychium species have suggested various medicinal applications. These attracted us to consider Hedychium species for our research. The plant's whole body, essential oil and solvent extracts from rhizomes are the main sources of biological activity. Compositions of essential oil obtained from dried rhizomes with good anti-inflammatory activity [1,[6][7][8], and extracts obtained from different parts (i.e., leaves, rhizomes, flowers of plant (as shown in Figure 1)) that contain numerous phytoconstituents (such as Hedychenone, Coronarin-D, Hedychilactone-D, Hedychinal and many more) that exhibit anti-inflammatory activity and other biological activity (such as antihistaminic, hair growth, skin care, and cytotoxic (in breast cancer) effects etc.) have also been reviewed several times. Therefore, herein we reviewed and discussed the current updated information found in our search of traditional applications and knowledge of the plant, emphasizing and aiming to identify its pharmacological applications for human use. For that purpose, details on ethnopharmacology, phytochemistry, extraction, purification, isolation, identification, characterization, and the pharmacological properties of the chemical constituents obtained from Hedychium species are under consideration.

Materials and Methods
An exhaustive literature search was accomplished by using different online search engines such as PubMed, PubChem, Sci-finder, ChemSpider, Science Direct, Mendeley, Sco- Table 1. Phytoconstituents and their semisynthetic derivatives from Hedychium species.

Chemical Structure
Phytoconstituents and Their Derivatives Chemical Nature Reference
Labdane diterpene: Cytotoxic compounds Coronarin A, Coronarin B, Coronarin C, and Coronarin D were isolated from Hedychium coronarium, also used in rheumatism in Brazil.

Saponins
Water and alcoholic extract passed a foaming test for Saponins, and a physiochemical test of Hedychium spicatum, a rhizome powdered drug [3] with individual compounds not yet identified.

Essential Oil
The physiochemical properties of essential oil obtained from Hedychium spicatum are given in Table 2. The oil contains alpha-pinene, beta-pinene, Limonene, 1:8 Cineole, Linalool found in major amounts, and Camphore, Linalyl acetate, Terpineol, Borneol, Caryophyllene, r-Cadinene, humulene, Terpineolene and P-Cymene in low quantities. These compounds were studied by TLC and GLC methods [8]. The chemical composition of the essential oil of Hedychium spicatum rhizomes by gas chromatography shows that the essential oil contains Caryophyllene, monoterpenes, sesquiterpenes, and sesquiterpene alcohol [8,36]. The essential oil was isolated from chopped rhizome with steam distillation, and the distillate was further saturated with NaCl, then further extracted with petroleum ether (60-80 • C) and hexane. Isolation of compounds from the essential oil at a boiling point of 60-80 • C yielded a (0.3%) compound using fractional distillation, with a spinning band distillation assembly based on the boiling point. 1,8-cineol (boiling point 62 • C), (+)-linalool (boiling point 120 • C), five alcohol, (+)-Elemol (26%), (−)-epi-10-Gama-eudesmol (19%), and (30%) of (−)-alpha-Cadinol, alpha-Eudesmol, (+)-beta-Eudesmol mixture were isolated. Bottini et al. reported the essential oil constituents isolated from different Hedychium species, which are listed in Figure 4 and Table 3.           129 Larixol is a Labdane isolated from Larch oleoresin, converted into Aldehyde using a 3-step transformation using osmium tetroxide/sodium periodate oxidation. Free alcohol is protected by silyl ether followed by elimination of tertiary acetate using 2,4,6-collidine, which is identified by the presence of vinylic protons. 3-furyl lithium is used for addition reaction to the aldehyde group and converted into a mixture of Epimeric alcohols. The Epimeric alcohols are mesylated in the presence of 2,6-lutidine, which produces an elimination product with trans configuration. Cleavage of silyl ether followed by oxidation produces an intermediate which produces Hedychenone on isomerization, and upon reduction with di-isobutyl, aluminum hydride yields Yunnacoronarin A. The reaction scheme is shown in Figure 5.

Synthesis of Hedychenone, Yunnacoronarin A from Larixol
Larixol is a Labdane isolated from Larch oleoresin, converted into Aldehyde using a 3-step transformation using osmium tetroxide/sodium periodate oxidation. Free alcohol is protected by silyl ether followed by elimination of tertiary acetate using 2,4,6-collidine, which is identified by the presence of vinylic protons. 3-furyl lithium is used for addition reaction to the aldehyde group and converted into a mixture of Epimeric alcohols. The Epimeric alcohols are mesylated in the presence of 2,6-lutidine, which produces an elimination product with trans configuration. Cleavage of silyl ether followed by oxidation produces an intermediate which produces Hedychenone on isomerization, and upon reduction with di-isobutyl, aluminum hydride yields Yunnacoronarin A. The reaction scheme is shown in Figure 5.

Synthesis of Yunnacoronarin D from Hedychenone
Yunnacoronarin D is synthesized from Hedychenone by oxidation of the allylic methyl group and reduction of the aldehyde group. Photo-oxidation of Hedychenone also produces Yunnacoronarin [33]; the reaction scheme is shown in Figure 5.

Synthesis of Yunnacoronarin D from Hedychenone
Yunnacoronarin D is synthesized from Hedychenone by oxidation of the allylic methyl group and reduction of the aldehyde group. Photo-oxidation of Hedychenone also produces Yunnacoronarin [33]; the reaction scheme is shown in Figure 5 they are Exo and Endo isomers. The Exo isomer is asymmetrically converted to Hedychenone by reduction, oxidation, olefination, and de-sialylation in the presence of 3-furyl ylide [42]. The Ester cycloadduct intermediate is reduced to give intermediate aldehyde, which reacts with tri-phenylphosphoranylidene lactone in methane dichloride to yield Hedychilactone B [21]; the reaction scheme is shown in Figure 6.
Hedychenone undergoes epoxidation with m-CPBA in DCM at room temperature. The synthesised compound was further characterized by NMR, Mass and FTIR. The SAR indicates that the Furanoid ring system has good cytotoxicity, rather than the Decalone nucleus. Dimerization through C-8 was found to significantly enhance the cytotoxic activity of Hedychenone. ranylidene lactone. (i), (ii), and (iii) are synthetic roots for the preparation of intermediates.

Other Reactions
Enzymatic hydrolysis of Coronalactosides I with Naringinase Coronalactoside-I in 0.1 M acetate buffer (pH 3.8, 1.0 mL) was treated with Naringinase solution at 40 • C for 24 h, thereby forming Coronalactone. Workup and chromatography was performed on the mixture over reverse-phase silica gel. Coronalactone was found to be a colourless oil, and [α] D -12.9 • . The reaction scheme is shown in Figure 8.
nucleus. Dimerization through C-8 was found to significantly enhance the cytotoxic activity of Hedychenone.

Other Reactions
Enzymatic hydrolysis of Coronalactosides I with Naringinase Coronalactoside-I in 0.1 M acetate buffer (pH 3.8, 1.0 mL) was treated with Naringinase solution at 40 °C for 24 h, thereby forming Coronalactone. Workup and chromatography was performed on the mixture over reverse-phase silica gel. Coronalactone was found to be a colourless oil, and [α]D-12.9°. The reaction scheme is shown in Figure 8. Labdane diterpenes Hedychenone was converted to Yunnacoronarin A, Yunnacoronarin D, Hedychilactone B, 6,7-Dihydrohedychenone, 6,7,11,12-Tetra hydro Hedychenone, Aldehyde product of Hedychenone, and Dimer of Hedychenone were found to be more cytotoxic than Hedychenone. Coronalactone had an active moiety of Coronalactoside, and hepatoprotective activity.

Herbal and Traditional Uses of Hedychium species
Various Hedychium species are used to treat ailments and diseases in traditional herbal medication. Extracts, decoctions, infusions, macerates, oils and squeezed liquid forms are used for different administrations, as listed in Table 4.

Herbal and Traditional Uses of Hedychium species
Various Hedychium species are used to treat ailments and diseases in traditional herbal medication. Extracts, decoctions, infusions, macerates, oils and squeezed liquid forms are used for different administrations, as listed in Table 4.

Pharmacological Activity of Hedychium species
Hedychium species possess various pharmacodynamic activities based on different activity that has been carried out by researchers. Table 5 contains the pharmacodynamic activities of the extract, and the chemical constituents present in extract.

Anti-Inflammatory and Analgesic Activity
In a personal communication report by Dhawan, B.N, (Pharmacology division, CDRI, Lucknow) it was reported that the ethanolic extract of the Hedychium spicatum plant rhizome has anti-inflammatory properties [31].   This test found that chloroform and methanolic extract of 400 mg/kg bw both inhibited writhing reflux (27.23% by chloroform and 40.59% by methanolic extract) in eight groups of pre-screened Swiss albino mice with significant p < 0.001 and 50 mg/body weight of aminopyrine as control. Inhibition was measured using formula [67]:

Radiant Heat Tail-Flick Method for Analgesic Activity
Tail-flick latency was assessed using morphine 2 mg/kg body weight as control and an analgesiometer. The percentage of elongation measures showed significant response for radiant heat, with p < 0.001 [67].

Carrageenan-Induced Rat Hind Paw Edema Animal Model for Anti-Inflammatory Study
This model was used to estimate acute inflammation. Different concentrations of hexane, chloroform, methanol extract were tested against PBZ (Phenylbutazone), with an 80 mg/kg dose and percentage inhibition calculated using the following formula: where V c and V t represent paw volume. The control study found that chloroform and the methanolic extract of Hedychium coronarium rhizome extract exert significant p < 0.01 inhibition of paw volume, at 27.46% and 32.39%, respectively, with 400 mg/kg body weight; meanwhile, the control, with 80 mg/kg, showed a 42.54% inhibition with a significance value of p < 0.001 [67].

• In Vitro Inhibitory Assay Oof NO Produced in LPS and IFN-g-Stimulated 264.7 Macrophages
Labdane diterpenes, Hedychenoids A, Hedychenoids B, Hedychenone, Forrestin A, and Villosin were isolated from the rhizomes of Hedychium yunnanense by ethanol maceration, and further purified by liquid-liquid extraction using ethyl acetate and water, then butanol. Hedychenoids B and Villosin had an inhibitory effect, with IC 50 values of 6.57 ± 0.88 and 5.99 ± 01.20 µg/mL respectively [85].

Inhibition of Acetic Acid-Induced Vascular Permeability in Mice (Anti-Inflammatory)
Histamine and serotonin play an important role in the vascular permeability induced by acetic acid, an exudative state of inflammation. The anti-inflammatory effects of methanolic extract of Hedychium coronarium and some labdane diterpenes (Coronarin D, Coronarin D methyl ether) were tested, and the study found that methanol extract (Dose 250-500 mg/kg), Coronarin D (Dose 25-50 mg/kg), and Coronarin D methyl ether (Dose 25-50 mg/kg) had a significance of p < 0.05-p < 0.01 [17].

Inhibition of Acetic Acid-Induced Vascular Permeability in Mice (Anti-Inflammatory)
Histamine and serotonin play an important role in the vascular permeability induced by acetic acid, an exudative state of inflammation. The anti-inflammatory effects of methanolic extract of Hedychium coronarium and some labdane diterpenes (Coronarin D, Coronarin D methyl ether) were tested, and the study found that methanol extract (Dose 250-500 mg/kg), Coronarin D (Dose 25-50 mg/kg), and Coronarin D methyl ether (Dose 25-50 mg/kg) had a significance of p < 0.05-p < 0.01 [17].

Antioxidant Activity
Chan et al., 2008 carried out estimation of total phenolic content and free radicalscavenging activities using a Folin-Ciocalteu, and DPPH radical scavenging assay. The

Antioxidant Activity
Chan et al., 2008 carried out estimation of total phenolic content and free radicalscavenging activities using a Folin-Ciocalteu, and DPPH radical scavenging assay. The methanolic extract of leaves of Hedychium coronarium (from the Lake Gardens of Kuala Lumpur, Malaysia) were used for the study. The methanolic extract obtained from rhizomes of Hedychium spicatum was found to have potent antioxidant activity [71,73,76]. The total phenolic content was estimated using the gallic acid calibration equation y = 0.0111 × 0.0148 (R 2 = 0.9998), and the total phenolic content of Hedychium coronarium was 820 ± 55 mg GAE/100 g, with significance of p < 0.05. The DPPH radical scavenging activity [88] was calculated as IC 50 and expressed as the ascorbic acid equivalent antioxidant capacity (AEAC), at IC 50 = 0.00387 mg/mL, whereas Hedychium coronarium had a capacity of 814 ± 116 mg AA/100 g, with significance of p < 0.05. AEAC (mg AA/100 g) = IC 50(ascorbate) / IC 50(sample) ·10 5 [58]. Essential oil from a Hedychium gardnerianum Sheppard ex Ker-Gawl leaf was evaluated for DPPH antioxidation activity, and good antioxidant activity was found against ascorbic acid and BHT, as standard [86].

Anti-Microbial Activity and Anti-Fungal Activity Anti-Microbial Activity and Anti-Fungal Activity by Disk Diffusion Method
Aqueous, methanol, ethanol, acetone, and hexane extracts of Hedychium spicatum rhizome were evaluated against B. subtilis, S. aureus, M. luteus, E. coli, A. flavus, A. fumigatus, M. gypseum, and C. albicans, and the zone of inhibition was recorded in (mm) at different doses (mg/disc). The results found that the extracts were active against B. subtilis, M. luteus, E. coli, A. flavus, and C. albicans, and not active against S. aureus, A. fumigatus, and M. gypseum [74]. The essential oil of Hedychium coronarium was obtained through hydro distillation, and the composition of the essential oil was identified by gas chromatography combined with mass spectroscopy with a flame ionization detector; the major components were B-pinene, eucalyptol, linalool, Coronarin-E, etc. The essential oil exhibited DPPH radical-scavenging activities, and also inhibited C. albicans and F. oxysporum [71,89].
Bisht, G.S et al., 2006 carried out an anti-microbial study for petroleum ether, benzene, chloroform, ethyl acetate, acetone, ethanol, aqueous extract, and essential oil from Hedychium spicatum rhizome. Dimethyl sulfoxide (DMSO) was used as a dilunt for the extract, and Tween-20 was used as the diluent for essential oil. Trichodermalignorum, were used. The study found that the concentrations of anti-microbial and anti-fungal compounds were 20 mg/disc for extract and 500 µL/disc for essential oil, respectively. Gentamycin (10 µg/disc), penicillin (10 unit/disc), Vancomycin (30 µg/disc), and Methicillin (5 µg/disc) were used as standard anti-microbial agents, and Cyclohexamide (30 µg/disc) was used as a fungicide, using a Petri dish diffusion/agar diffusion test [75].
S. Joshi et al. carried out an antimicrobial assay using a Petri dish diffusion method, testing the rhizome essential oil (50 µL/disc) of Hedychium ellipticum, Hedychium aurantiacum, hedychium coronarium, and Hedychium spicatum and control Amikacin, Ciprofloxacin, Ampicillin, Gentamycin, and Tetracycline on bacterial strains S. aureus, Sal. Enterica, Pasteurella multocida, Shigella flexneri and Escherichia coli. The minimum inhibitory concentration of essential oil was found to range from 0.97 to 62.5 µL/mL, depending on the susceptibility of the tested organism [72].
In Vitro Antimicrobial Activity Using Agar Diffusion/Disk Diffusion Method A Mueller-Hinton agar plate was cultured with microbial broth culture to study the zone of inhibition, using Hedychium coronarium rhizome essential oil (Contains Monoterpenes, Di-terpenes, and sesqui-terpenes), by cylinder plate method. The plates were incubated for 24 h at 37 • C for bacteria and 24-48 h at 28 • C for fungi. The bacteria Bacillus subtilis and Pseudomonas aeruginosa and the yeast-like fungus Candida albicans and Trichoderma sp. (Dermatophyte) were studied. The study found that the essential oil of rhizomes has a better inhibitory effect against Trichoderma sp. and Candida albicans than against Bacillus subtilis and Pseudomonas aeruginosa [41].

Caenorhabditis Elegans Mobility Test for Anthelmintic Activity
Lima A et al., 2021 carried out anthelmintic activity using adult C. elegans, both susceptible (wild-type and Bristol N2), and Ivermectin-resistant; a balance saline solution was used in 24-well plates for treatments with nematodes. The concentration of Hedychuim coronarium rhizome essential oil and standard monoterpenes, i.e., α-Pinene, β-Pinene, (S)-(−) Limonene, (R)-(+) Limonene 1,8-Cineole, and p-Cymene, was found to range from 0.009 to 10 mg/mL, diluted in DMSO 1%. After 24 h at 24 • C, mortality was evaluated with a negative control mixture of M-9 solution and Ivermectin as positive control. The study found that the essential oil of Hedychium coronarium rhizomes had an IC 50 concentration of 0.082 mg/mL (IC 95 concentration 0.058-0.117 mg/mL) for the Bristol N2 strain, and an inhibitory concentration of IC 50 of 0.82 mg/mL (IC 95 0.556-1.2 mg/mL) for the Ivermectinresistant strain, with a significance value of p < 0.05 [40].

Anti-Histaminic, Mast Cell-Stabilizing and Bronchodilator Effect
An in vitro study of hydroalcoholic extract of root composition containing Hedychium spicatum root was carried out to examine its antihistaminic effect (with Histamine dihydrochloride as control) on Guinea pigs, with a composition of 50 mg/kg of Hydroalcoholic extract. It effectively works as a preventive-type antagonist. When investigating mast cell stabilization (Ketotifen fumarate as control) on rats, a 1000 µg/mL concentration composition produced 54-58% inhibition of mast cell degranulation, with a significance value of p < 0.001. The bronchodilatory effect of the extract composition on histamine-induced bronchospasm (80-86%) was investigated in guinea pigs; a 200 mg/kg-500 mg/kg composition increased pre-convulsion time by 27-36%, with a significance value of p < 0.001 [84].

In Vitro Cytotoxicity Assay
Labdane diterpenes, Hedychenoids A, Hedychenoids B, Hedychenone, Forrestin A, and Villosin were isolated from the rhizomes of Hedychium yunnanense by ethanol maceration, and further purified by liquid-liquid extraction using Ethyl acetate and water, then Butanol. The compounds were tested using an SRB method on SGC-7901 (the human gastric cancer cell line) and HELA (human cervical carcinoma). The study found that Hedychenoids B, Hedychenone, and Villosin had cytotoxicity against SGC-7901, with IC 50 values of 14.88 ± 0.52, 7.08 ± 0.21 and 7.76 ± 0.21 µg/mL, and against HELA, with IC 50 values of 9.76 ± 0.48 and 13.24 ± 0.63 µg/mL, respectively [85].

Ameliorating Potential
The protective effect of Hedychium spicatum rhizome powder with concentrations of 4000 ppm and 2000 ppm was tested against 250 ppm IC 50 of Indoxacarb-induced toxicity on a group of cockerels. The ameliorative effect of Hedychium spicatum root powder and its ability to restore the gene activities and expression of antioxidant, biotransformation, and immune system genes were demonstrated in cockerels fed Indoxacarb [77,93,94]. The study concluded that 80% Aqueous acetone extract of Hedychium coronarium flower and other chemical constituents had a hepatoprotective effect, with a significant p < 0.01 value of percentage inhibition [25]. The expression of hepatic genes associated with biotransformation, antioxidant, and immune systems in WLH cockerels fed indoxacarb was evaluated, and a protective effect of Hedychium spicatum root extract was found. The extract prevents changes in expression of antioxidant, biotransformation, and immune system genes [65]. Figure 10 shows inhibition of D-GalN-induced cytotoxicity. The hydro distillate essential oil of Hedychium spicatum rhizomes has been tested on P.humanuscapitis (Phthiraptera: Pediculidae). Essential oil of 1, 2 and 5% concentration was blended with coconut oil as a base. A 1% permethrin-based preparation was used as a positive control. Lice clasping hair strands were immersed completely in the test solutions and the marketed preparation for 1 min. Vital signs were measured after 5,10,15,20,30,45,60,90, and 120-min; lice were judged to be dead if a vital sign was zero. Mortality was observed as 85%, 80%, and 75% for the 5%, 2%, and 1% Hedychium spicatum essential oil; after 2 h, 100% mortality was observed, which is as significant as 1% permethrin preparation [78].

Hair-Growth Promotion Activity
Pentadecane and Ethyl para methoxy cinnamate were isolated from Hedychium spicatum rhizome hexane extract and evaluated for the in vivo hair growth promotion activity on female Wistar rats weighing 120-150 g. The results found that pentadecane demonstrates good reduction in hair growth time, but hexane extract shows better-than-individual compound activity [30].

CNS Depressant Activity of Hedychium Spectrum Extract on Rats
Ethanolic, hexane, and chloroform extracts were evaluated at a dose of 100 mg/kg body weight for CNS activity, and it was found that the extracts have CNS depressant activity using Gabapentin and Caffeine 250 mg/kg body weight as control [79].

Skin Protective Composition
A composition of Hedychium extract has been reported to treat environmental damage to the skin, regulating firmness, tone, wrinkles, and skin texture with a cosmetically acceptable carrier.

Inhibition of UV-Induced Matrix Metalloproteinase-(MMP)
Epidermal equivalents derived from human epidermal keratinocytes topically treated with 0% or 0.5% w/w of Hedychium spicatum were irradiated with solar spectrum light and analysed with ELISA, which proved that compositions containing Hedychium spicatum extract provide protection against UV-induced matrix mettalloproteinase-1 (MMP-1) (the UV light in 15 MED's effect on MMP-1: 19.3 pg/mL was reduced to 11.2 pg/mL with 0% Hedychium extract, and on MMP-1: 31.2 was reduced to 2.1 pg/mL composition with 0.5% Hedychium extract) [80].

Prevention of Smoke-Induced Loss of Thiols in Normal Human Dermal Fibroblasts
Glutathione works as a redox buffer by maintaining a balance of oxidants and antioxidants. UV exposure depletes antioxidants and glutathione, which leads to higher UVR sensitivity that causes wrinkling on the skin and environmental damage. Ten minutes of exposure to smoke reduces the thiols percentage, hence a 100 µg/mL Hedychium spicatum extract concentration afforded thiols protection in 106 ± 15.8 (mean ± SD) smoke groups [80].

Inhibition of Nitric Oxide Production
The ability of Hedychium extract to inhibit nitric oxide production was tested in LPSstimulate murine macrophages. Nitric oxide is involved in physiological processes such as vasodilation, neurotransmission, inflammation, and growth of cancers. Nitric oxide combined with one superoxide radical produces peroxy nitrite, a highly toxic free radical. Murine macrophage RAW 264.7 was treated with Hedychium extract with concentrations of 10 to 200 µg/mL, and lipopolysaccharide from E. coli. with an IC 50 of 69.97 µg/mL [80].

Compositions Used to Darkenthe Skin
The composition of Hedychium extract, some peptides and other extracts were tested for their ability to darken to skin and have been studied in in vitro and in vivo on human skin.

Induced Pigmentation in Human Cell Culture
Keratinocyte-melanocyte culture (Human HaCaT keratinocytes) was used to test different peptide (at 50 µM), pigment melanin and derivatives of melanin (0.0001% w/v t 1% w/v), and plant extract Forskolin was used as a pigmentation inducer. L-3,4-Dihydroxyphenylalanine (DOPA) staining and computerized image analysis were carried out (the parameters measured were the surface area of stained material within the melanocytes and keratinocytes, the total surface area of cells in culture, and the related pigmented area). Cell viability was assayed using Alamar Blue TM , and the increased pigmentation level of a peptide + Hedychium extract (50 µM, 0.1% w/v) composition was tested during analysis; the mean related pigmented area was 0.050, and the control mean area was 0.150 [81].

Induced Pigmentation In Vivo
Dark-skinned Yucatan micro swine were used for analysis of the increase in pigment deposition Forskolin or Coleus extract. Versus a positive control (1% w/v); pigment increased from 1% to 5% w/v, and peptide from 250 µM to 500 µM. Hedychium spicatum extract dissolved in ethanol: propylene glycol at a ratio of 70:30 v/v indicated a strong increase in pigment deposition, and some increase in caps [81].

Darkening of Human Skin
Human skin was tested (after being obtained from patients undergoing cosmetic surgery). Human graft was treated with composition of peptide (500 µM) and soluble melanin Melasyn-100 tm (1% w/v) in Lysosome (20 mg/mL) as a control; histological sections were evaluated for changes in pigment deposition and capped epidermal cells above the basal layer [81].
The effect of ethanol extract of leaves and pseudo stem of Hedychium coronarium on melanogenesis in B16 cells was evaluated (through melanin titration); stimulation of melanin release was inhibited by the extract at 1 mg/mL concentration. This indicates that it may help to inhibit sun-induced pigmentary spots [96].

Composition Containing Active Sunscreen Agent
Hexane and ethyl acetate fractions were isolated over silica gel and 60-120 mesh containing p-Methoxy cinnamic acid esters from Hedychium spicatum extract. Formulations containing Hedychium spicatum extract were analyzed for SPF 13.97 using an SPF 290S analyzer; the formulations were sun protection cream, sun protection shampoo, and sun protection gel, all containing Cinnamic acid ester active fractions. Skin irritation was tested in Guinea pigs, and localized reversible dermal responses were checked without the involvement of immune response [99].

Composition Treating Tenia Infection
Hedychium spicatum extract prepared by pulverized rhizome was extracted with chloroform at 60 • C. 6.5% w/w. Ethyl-p-methoxycinnamate was extracted and isolated by silica gel column chromatography using pet ether in ethyl acetate. Final crystallization was carried out using pet ether. An M.P of 48-50 • C was characterized by 1 H NMR and MS 206 (M+) [82,100].

Antidiabetic Activity of Extract and Composition Used to Reduce Blood Glucose Level
Ethanol extract of the leaves and pseudo stem of Hedychium coronarium was tested for glucose tolerance in normal rats (with a dose of 750 mg/kg reducing blood glucose in 120 min) and mice with type-II diabetes (with a dose of 1.5 g/kg causing a significant reduction in blood glucose, with a significance value of p < 0.01). An intraperitoneal glucose tolerance test was carried out in normal mice (with a dose of 1.5 g/kg, with p < 0.01), while an insulin increase test was carried out in normal mice (with a dose of 1.5 g/kg, with p < 0.01 value). A glucose tolerance test was carried out in rats with type-I diabetes (with a dose of 1.5 g/kg, with p< 0.01), and an insulin increase test was performed on mice with type-II diabetes (with a dose of 1 g/kg, with p < 0.01). An insulin resistance test was carried out in rats with type-I diabetes (with a dose of 1 g/kg, with p < 0.01) [64]. The water extract of leaves and the pseudo-stem of Hedychium coronarium were tested for glucose tolerance in normal rats (with a dose of 1.5 g/kg, with p < 0.01), and water-ethanol extract was tested for glucose tolerance in normal rats (with a dose of 0.8 g/kg, with p < 0.01) [64,101,102].

Anti-Inflammatory Composition in Cream
Inflammatory cytokines (TNF-alpha, IL-6, and IL-1beta in pcg/mg protein) were tested in the blood serum of mice by an enzyme-linked immune-sorbent assay (ELISA), which showed the topical synergistic anti-inflammatory activity of three essential oil blends, including 60% Cymbopogon citratus oil, 20% Zenthoxylumarmatum oil, and 20% Hedychium spicatum oil, which is beneficial in inflammatory arthritis [103]. The macrobiotic composition of Hedychium spicatum extract, along with other ingredients, shows health benefits that are effective in facial and body care for acne, dermatitis, eczema [104], and conditioning of the skin [105]. Various extract-and essential oil-based formulations have been produced for the treatment of immune system disorders such as cancer and lupus [106].

Uses in Cracked Heels
A cracked heel cream composition containing Hedychium spicatum extract may be effective as an anti-inflammatory and used as barrier to protect the skin [83].
2.6.13. Therapeutic Effect of Composition of Plant Extract of Hedychium Coronarium Root for Treatment of the Human Body A mitochondrial network in fibroblasts was irradiated with UVA (Mito-tracker staining, ATP, and NAD+/NADH titration). Cytotoxicity and viability were checked by an XTT assay. Concentrations of 0.005% and 0.01% extract were non-cytotoxic to fibroblasts [96]. The lysosomal network in fibroblasts was irradiated with UVA (Lysotracker staining).
Evaluation of the effect of the root extract on human skin explants was then performed using a pollution model. The extract has the potential to fight against inflammatory stress mediators induced by pollution [100]. The effect of β-endorphin production by normal human keratinocytes was evaluated; treatment of normal keratinocytes with extract concentrations of 0.001 to 0.005% induced stimulation of β-endorphin release [96].
The antioxidant potential of the ethanol extract of the leaves and pseudo stem of Hedychium coronarium was evaluated in normal human keratinocytes (using ROS detection with an H2DCFDA probe). The extract showed inhibitory action on ROS production with a concentration of 0.01%, p < 0.122 [96]. The effect of the ethanol extract of leaves and the pseudo stem of Hedychium coronarium on autophagic activity in fibroblasts was evaluated by irradiation with blue light (according to MDC assay, the autophagic activity of fibroblast cells decreases by 13-35% at concentrations of 0.001% to 0.005%) [96].

Discussion
Hedychium plants are easily available in the Himalayan regions of India and China [44,52]. Hedychium plants are used in gardening and for some traditional applications. Utilization of plants for suitable and precise medicinal activity is needed. In our study, we found Hedychium species are abundant with terpenes and terpenoids. Diterpenes and diterpenoids i.e., Hedychenone, Hedychilactone D, Coronarin D, Coronarin D ethyl ether etc. [9,32] may have future potential as anti-inflammatories, and may be effective against inflammatory mediators and precursors. Terpenoids are hexane-soluble, while hexane extracted materials are not likely to be used for human intake. Suitable green technology may help the extract to become more prominent in human uses, such as super critical fluid extraction, and ethanol-based extraction methods. Extracts and individual compounds were isolated from Hedychium spicatum and Hedychium coronarium, and both species were studied in vitro for anti-inflammatory, antifungal, antimicrobial, and cytotoxic qualities [44,45,69].
Identification of possible pharmacological pathways and the specific medicinal activity of individual drug candidates may help to improve and strengthen plant-based herbal API (active pharmaceutical ingredients). Screening of pharmacological activity using artificial learning/machine learning (AL/ML) methods [107] helps to draw conclusions on the specificity of a given compound. Computational approaches such as docking, MD simulation, virtual screen for different and most suitable protein-binding sites, and ADMET study [108] for stabilizing pharmacokinetic parameters may be used To increase compounds' potency and receptor specificity, de novo drug design [109] and receptor-based drug design approaches may be used.

Conclusions
Terpenes are used in perfumery and aromatherapy due to their pleasant odor and aromatic effect. This study found that Hedychium species are abundant with aromatic properties. Hedychium species contain monoterpenes and sesquiterpene present in the essential oil of leaves, flowers, rhizomes, and roots. Terpenes are chiral in nature, meaning they provide distinct physiological characteristics to the compounds, such as odor, medicinal activity, and toxicity. Unsaturation is a key feature of terpenes, as is the presence of oxygen atoms in terpenoids. Hexane, ethyl acetate, and chloroform extracts of plant parts of the Hedychium species contain various diterpenes, diterpenoids, and labdane-type diterpenoids. Methanol, ethanol, hydroalcoholic, and aqueous extract fractions contain polyphenolic compounds, flavonoids, xanthones, and some glycosides.
The essential oil of Hedychium plant parts contains aromatic, anti-inflammatory, and antimicrobial activity due to mono and sesquiterpenes. Hexane, ethyl-acetate and chloroform fractions exhibit significant anti-inflammatory, cytotoxic, antimicrobial activity due to terpenoids and labdane diterpenes. The methanol and ethanol extracts of Hedychium plant parts contain antioxidant and hydroalcoholic extract and possess antioxidant and bronchodilator activity. Herbal ancient medication using the different parts (e.g., leaves, rhizomes, and flowers) of Hedychium extracts has effective anti-inflammatory, analgesic, antidiabetic, and anti-asthmatic qualities; it is also used as an antidote to snake bites, and in various other synergistic activities. This is because in traditional medication systems, the plant's whole parts, dried powder and hydroalcoholic or alcoholic extract are used, which contain all the active molecules needed to treat a disease or ailment.
Our study suggests that the extracts and essential oils of Hedychium spicatum, Hedychium coronarium, Hedychium ellipticum, Hedychium aurantiacum, Hedychium gardnerianum have anti-inflammatory, anti-pyretic, skin protective (via sunscreen), and anti-bacterial effects. The chemical constituents present in hexane, chloroform, and ethyl acetate, and in the methanolic extract of Hedychium spicatum and Hedychium coronarium, i.e., Hedychenone, Hedychilactone D, Coronarin D, Coronarin E, 9-Hydroxy Hedychenone, 7-Hydroxy Hedychenone, Yunnacoronarin A and Coronarian D methyl ether, Coronarian D ethyl ether (diterpenes and diterpenoids), are identified as having anti-inflammatory, anti-allergic, antibacterial, and cytotoxic effects. The study comprises useful findings regarding the extracts and isolated compounds of Hedychium species, which may be fruitful in the commercialization of specific compounds or extracts obtained from the Hedychium genus.