Galangal-Cinnamon Spice Mixture Blocks the Coronavirus Infection Pathway through Inhibition of SARS-CoV-2 MPro, Three HCoV-229E Targets; Quantum-Chemical Calculations Support In Vitro Evaluation.

Natural products such as domestic herbal drugs which are easily accessible and cost-effective can be used as a complementary treatment in mild and moderate COVID-19 cases. This study aimed to detect and describe the efficiency of phenolics detected in the galangal-cinnamon mixture in the inhibition of SARS-CoV-2's different protein targets. The potential antiviral effect of galangal-cinnamon aqueous extract (GCAE) against Low Pathogenic HCoV-229E was assessed using cytopathic effect inhibition assay and the crystal violet method. Low Pathogenic HCoV-229E was used as it is safer for in vitro laboratory experimentation and due to the conformation and the binding pockets similarity between HCoV-229E and SARS-CoV-2 MPro. The GCAE showed a significant antiviral effect against HCoV-229E (IC50 15.083 µg/mL). Twelve phenolic compounds were detected in the extract with ellagic, cinnamic, and gallic acids being the major identified phenolic acids, while rutin was the major identified flavonoid glycoside. Quantum-chemical calculations were made to find molecular properties using the DFT/B3LYP method with 6-311++G(2d,2p) basis set. Quantum-chemical values such as EHOMO, ELUMO, energy gap, ionization potential, chemical hardness, softness, and electronegativity values were calculated and discussed. Phenolic compounds detected by HPLC-DAD-UV in the GCAE were docked into the active site of 3 HCoV-229E targets (PDB IDs. 2ZU2, 6U7G, 7VN9, and 6WTT) to find the potential inhibitors that block the Coronavirus infection pathways from quantum and docking data for these compounds. There are good adaptations between the theoretical and experimental results showing that rutin has the highest activity against Low Pathogenic HCoV-229E in the GCAE extract.

Evaluation of Antifungal Activity of Languas galangal Rhizome and Development of a Topical Antifungal Cream.

Background: The rhizome of Languas galangal is traditionally used in Sri Lanka for the treatment of skin infections caused by fungi. The aim of the present study was to evaluate the antifungal activity of L. galangal rhizome and to develop a topical antifungal formulation from it. Methods: The dried, powdered rhizome of L. galangal was successively extracted with hexane, dichloromethane, ethyl acetate and methanol using Soxhlet extraction. The agar well diffusion method was used to assess the antifungal activity against Candida albicans and Aspergillus nger. The antifungal activities of the extracts were compared with clotrimazole as the positive control and dimethyl sulfoxide (DMSO) as the negative control. The most active hexane extract was used to prepare the cream. The antifungal activity of the formulated cream was tested. Results: The hexane extract of L. galangal rhizome powder was more effective on C. albicans and A. niger. The hexane extract of L. galangal showed the maximum zone of inhibition against C. albicans and A. niger (20.20 mm ± 0.46, 18.20 mm ± 0.46) compared to the other three extracts, while clotrimazole, which was used as a positive control, produced a larger zone of inhibition (36.10 mm ± 0.65) and dimethyl sulfoxide (DMSO), the negative control, did not produce inhibitory zones. Stability testing of the formulated cream showed a stable and good appearance. Conclusions: The cream developed using the hexane extract showed in vitro antifungal activity against C. albicans and A. niger. Further evaluations on shelf life, stability and safety are required.

Lactating buffalos' productive performance, ruminal kinetics, nutrients digestibility and oxidative status as response to supplementing diets with alpinia galanga.

The current study investigated the effect of adding different doses of galangal rhizome powder (Alpinia galanga) to the diets of lactating buffaloes on milk production, composition and in vitro ruminal fermentation kinetics. Twenty buffaloes were divided into four groups: (T1) a control diet comprising of concentrates, corn silage and rice straw at 50: 35: 15 on a DM basis, (T2) a control diet plus 15 g of galangal/animal/day and (T3) a control diet plus 30 g of galangal/animal/day and (T4) a control diet plus 45 g of galangal/animal/day. Results showed that inclusion of galangal in diets decreased (p < 0.05) the concentrations of butyrate, ammonia-nitrogen, CO2, and total protozoa count as compared with T1. Acetate concentration was lower in T4 as compared with other groups (p < 0.05). While propionate concentration was lower (p < 0.05) in T2 as compared with T1. T3 and T4 lowered (p < 0.05) methane, CO2 productions, and protozoa count. T3 increased (p < 0.05) milk yield, energy corrected milk (ECM), yield of total solids, total protein, and solids not fat (SNF) than recorded for T1. While T4 recorded the lowest values (p < 0.05) of milk yield, ECM, total solids, fat, and total protein. The current results support the hypothesis that the inclusion of galangal in lactating buffaloes' diets at 30 g per animal daily improved feed intake and milk yield.

Pectin-type polysaccharide from galangal: An efficient emulsifier to construct the emulsion-based delivery system for galangal flavonoids.

Galangal is rich in flavonoids and polysaccharides but underutilized. In this study, galangal flavonoids and polysaccharides (GP-HN and GP-UN) were obtained by segmented extraction, used for chemical composition determination/structural characterization, and constructed for the emulsion delivery system. The results showed that galangin accounted for 71.45 % of total flavonoids. GP-HN and GP-UN were prepared by enzymatic-assisted high-temperature and ultrasonic extraction, which were low-molecular-weight pectin-type polysaccharides mainly constructed by galacturonic acid, galactose, and arabinose. GP-UN was the best emulsifier due to interfacial activities, emulsifying properties, interfacial resistance to bile salts displacement abilities, and anti-lipid digestion abilities of GPs. GP-UN emulsion could stably deliver flavonoids. This study presented a method for orderly reorganizing flavonoids and polysaccharides, guiding for utilization of whole bioactive components in galangal.

Pulsed electric field energy calculation to damage red galangal (Alpinia purpurata, K. Scumm) rhizome slices and its essential oil yield and quality with hydrodistillation.

This study aimed to determine the amount of energy to damage the red galangal rhizome sliced cell tissue and the amount and quality of the essential oil obtained by steam-water distillation. This study was a randomized block design, with pulsed electric field (PEF) voltage treatment starting at 1000, 2000, 3000, 4000, and 5000 V and without PEF and repeated three times. The results showed that the voltage between 3000 and 4000 V (E = 120-160 V/cm) or equivalent to 271.5-365.0 kJ/cm3 had damaged the red galangal rhizome slice cell tissue. The increase in yield due to PEF ranged from 13% to 73%, and there was no change in the value of the refractive index and specific gravity, but there was a change in the chemical composition of the essential oil constituents. The benefits of research with PEF treatment are the increase in yield and shorter distillation time.

Stability Study of Alpinia galanga Constituents and Investigation of Their Membrane Permeability by ChemGPS-NP and the Parallel Artificial Membrane Permeability Assay.

Alpinia galanga Willd., greater galangal, has been used for thousands of years as a spice as well as in traditional medicine. Its central nervous system (CNS) stimulant activity and neuroprotective effects have been proved both in animal models and human trials. However, the compounds responsible for these effects have not been identified yet. Therefore, the main constituents (p-OH-benzaldehyde (1), trans-p-coumaryl-alcohol (2), p-coumaryl-aldehyde (4), galanganol A (5), galanganol B (6), trans-p-acetoxycinnamyl alcohol (7), 1'S-1'-acetoxychavicol acetate (ACA, 9), and 1'S-1'-acetoxyeugenol acetate (AEA, 10)) were isolated to investigate their aqueous stability and passive diffusion across the gastro-intestinal tract (GIT) membrane and the blood-brain barrier (BBB) by the parallel artificial membrane permeability assay (PAMPA). Our positive results for compounds 1, 2, 4, 7, 9, and 10 suggest good permeability, thus potential contribution to the effects of greater galangal in the CNS. The results of the PAMPA-BBB were corroborated by in silico chemography-based ChemGPS-NP framework experiments. In addition, examination of the chemical space position of galangal compounds in relation to known psychostimulants revealed that all the molecules in proximity are NET/SERT inhibitors. As ACA and AEA did not show much proximity to either compound, the importance of further investigation of their degradation products becomes more pronounced.

Acaricidal potential of active components derived from Alpinia galanga rhizome oils and their derivatives against Haemaphysalis longicornis (Acari: Ixodidae).

Acaricidal activities and acetylcholinesterase (AChE) inhibitory activities were evaluated of active constituents of the essential oil extracted from Alpinia galanga rhizomes cultivated from India and their derivatives against Haemaphysalis longicornis nymphs. In addition, the effect was investigated of active components of A. galanga oil on egg laying of adult females of H. longicornis and egg hatchability. Of the volatile components identified in A. galanga oil, ethyl cinnamate, ethyl methoxycinnamate, and methyl cinnamate at 0.32 mg/cm2 resulted in 100% mortality, respectively, indicating that the acaricidal activity of the A. galanga oil against H. longicornis nymphs could be attributed to these compounds. To evaluate the structure-activity relationship between cinnamate derivatives and their acaricidal activities, allyl cinnamate, benzyl cinnamate, isopropyl cinnamate, isobutyl cinnamate, and isoamyl cinnamate were selected. Among cinnamate derivatives tested, allyl cinnamate exhibited the most potent toxicity (LC50 = 0.055 mg/cm2) against H. longicornis nymphs. The allyl cinnamate was also tested for AChE activity in vivo in H. longicornis nymphs and was found to affect the AChE activity. Allyl cinnamate at 10-50 mg/mL inhibited egg laying of adult females of H. longicornis by 10-43%. Egg hatching was suppressed completely by treatment with allyl cinnamate at 50 mg/mL, whereas allyl cinnamate was minimally toxic against non-target earthworms, Eisenia fetida. These results suggest that allyl cinnamate can be used as an active ingredient for the development of eco-friendly tick acaricides against H. longicornis, a vector for Sever fever with thrombocytopenia syndrome (SFTS) virus.

Quantification of Flavonoids in Alpinia officinarum Hance. via HPLC and Evaluation of its Cytotoxicity on Human Prostate Carcinoma (LNCaP) and Breast Carcinoma (MCF-7) Cells.

BACKGROUND: Various plant species have been shown to be effective in the prevention or adjuvant therapy of cancer. Alpinia officinarum and its main phytochemicals have also been the subject of several studies for their anticancer properties. OBJECTIVE: The objective of this study is to analyze the extracts of A. officinarum to quantify flavonoids and to evaluate the growth inhibitory effects of the extracts on MCF-7 and LNCaP cells. METHODS: A. officinarum aqueous and hydroalcoholic extracts were analyzed by using High-Performance Liquid Chromatography (HPLC) for the quantification of three flavonoid compounds. Then, MCF-7, LNCaP, and fibroblast cells were treated with several concentrations (25, 50, 100, 200, and 400 µg/mL) of extracts (24, 48 and 72h). Cell viability was assessed using an MTT assay. Flow cytometry was conducted to evaluate apoptosis. RESULTS: Galangin and kaempferol (3.85 and 1.57 mg/g dry extract) were quantified, respectively, in hydroalcoholic and aqueous extracts using a validated method. The hydroalcoholic extract significantly decreased the viability of MCF-7 (IC50: 43.45µg/mL for 48h) and LNCaP cells (IC50: 168 µg/mL for 48h). The aqueous extract reduced cancer cell viability by more than 50% only at 200 and 400 µg/mL (72 h). Treatment of primary fibroblasts with both extracts showed no significant decrease in cell viability (25-100 µg/mL; 24 and 48h). The hydroalcoholic extract induced a significant increase in apoptotic cells in both MCF-7 and LNCaP cells. CONCLUSION: Obtained results demonstrated the cytotoxicity of A. officinarum through apoptosis induction in two cancer cell lines. Further investigations are required to determine the underlying apoptotic cell death mechanisms induced by A. officinarum in cancerous cells.

Effect of Galangal Essential Oil Emulsion on Quality Attributes of Cloudy Pineapple Juice.

Galangal essential oil is obtained from the rhizomes of galangal with proven anti-inflammatory, antioxidant, antiviral, and antimicrobial properties, which are valuable in the food industry. To explore the effect of galangal essential oil on the quality of pineapple juice, 0.05, 0.1, 0.2, and 0.4% galangal essential emulsion were added, and their influence on the physical stability, physicochemical properties, microbial quantity, and aroma profiles of cloudy pineapple juice were evaluated. The essential oil emulsion of galangal is a milky white liquid with a strong aroma of galangal. The pH values of emulsion increased from 4.35 to 5.05 with the increase in essential oil concentration, and there was no significant difference in the particle size of the pineapple juice. The results showed that the galangal essential oil emulsion was stable and the stability of the cloudy pineapple juice was significantly enhanced by the essential oil emulsion determined using LUMiSizer. The cloudy pineapple juice with a 0.2% essential oil emulsion showed the most stability during storage. The lightness of the cloudy pineapple juice increased instantly with the essential oil emulsion addition. In addition, the microbial quantity of the cloudy pineapple juice was decreased by the individual essential oil emulsion or combined with thermal treatment to hold a longer shelf life. The microbial counts in pineapple juice treated by 0.4% essential oil emulsion and thermal treatment only increased from 1.06 to 1.59 log CFU/ml after 4 days of storage at 25°C. Additionally, the pH and total soluble solids showed a slightly increasing trend; however, the value of titratable acidity, free radical scavenging capacity, and ascorbic acid content of the cloudy pineapple juice showed no significant change. Finally, the results of the electronic nose showed that the aroma components of the pineapple juice were changed by the essential oil emulsion and thermal treatment, and the difference was especially evident in the content of the sulfur, sulfur organic, and aromatics compounds. Consequently, the results indicated that galangal essential oil emulsion can be used as juice additives to improve the quality attributes and extend the shelf-life of cloudy pineapple juice.

Comparative Study of Chemical Composition and Antioxidant Activity of Essential Oils and Crude Extracts of Four Characteristic Zingiberaceae Herbs.

The ginger family (Zingiberaceae) includes plants that are known worldwide to have a distinctive smell and taste, which are often used as spices in the kitchen, but also in various industries (pharmaceutical, medical, and cosmetic) due to their proven biological activity. The aim of this study was to investigate and compare the chemical composition and antioxidant activity (AA) of essential oils (EOs) of four characteristic ginger species: Elettaria cardamomum L. Maton (cardamom), Curcuma Longa L. (turmeric), Zingiber Officinale Roscoe (ginger), and Alpinia Officinarum Hance (galangal). Furthermore, the total phenolic content (TPC) and AA of crude extracts obtained after using ultrasound-assisted extraction (UAE) and different extraction solvents (80% ethanol, 80% methanol and water) were evaluated. A total of 87 different chemical components were determined by GC-MS/MS in the EOs obtained after hydrodistillation, 14 of which were identified in varying amounts in all EOs. The major compounds found in cardamom, turmeric, ginger, and galangal were α-terpinyl acetate (40.70%), ß-turmerone (25.77%), α-zingiberene (22.69%) and 1,8-cineol (42.71%), respectively. In general, 80% ethanol was found to be the most effective extracting solvent for the bioactivities of the investigated species from the Zingiberaceae family. Among the crude extracts, ethanolic extract of galangal showed the highest TPC value (63.01 ± 1.06 mg GA g-1 DW), while the lowest TPC content was found in cardamom water extract (1.04 ± 0.29 mg GA g-1 DW). The AA evaluated by two different assays (ferric-reducing antioxidant power-FRAP and the scavenging activity of the cationic ABTS radical) proved that galangal rhizome is the plant with the highest antioxidant potential. In addition, no statistical difference was found between the AA of turmeric and ginger extracts, while cardamom rhizome was again inferior. In contrast to the crude extracts, the EOs resulted in significantly lower ABTS and FRAP values, with turmeric EO showing the highest AA.

Carboxymethyl chitosan-pullulan edible films enriched with galangal essential oil: Characterization and application in mango preservation.

This study aimed to develop an active edible film based on carboxymethyl chitosan (CMCS) and pullulan (Pul) incorporated with galangal essential oil (GEO) by the casting method. And their physical properties, structural and preservation effect on mangoes were characterized. The CMCS/Pul ratio was determined to be 2.5:2.5 after the optimization of physical properties, mechanical properties and barrier properties of the blend film. The results of FT-IR and XRD showed that hydroxyl groups of Pul interacted with the carboxyl groups of CMCS and the blend films had good compatibility. Good thermal stability of CMCS/Pul-GEO films was further proven by TGA curves. The CMCS/Pul-8 %GEO film showed effective preservations on mango fruits during 15 days of storage at 25 ± 1 °C, based on the characterization by fruits weight loss, firmness, titratable acidity, soluble solids. Consequently, CMCS/Pul-GEO blend films may be a promising eco-friendly packaging material for the industrial application of fruit preservation.

A Randomized Placebo Controlled Clinical Trial Demonstrating Safety & Efficacy of EnXtra® in Healthy Adults.

Objective: The present randomized, placebo-controlled study aimed to assess the long-term safety and perceivable mental acuity benefits of EnXtra® in healthy individuals.Methods: Study participants were administered EnXtra® with or without caffeine for a period of 12 weeks. The cardiovascular safety was evaluated by assessing change in QT interval, blood pressure and heart rate. Further, other efficacy variables evaluated were change in perceived alertness and calmness by Bond and Lader mood scales, Sleep disturbance by Pittsburgh sleep quality Index and daytime sleepiness by Epworth sleepiness scale.Results: None of the study group showed any significant change in the ECG or haemodynamic parameters as compared to baseline (p > 0.05). Post consumption, alertness and calmness scores were significantly increased in the EnXtra®, and EnXtra® plus caffeine group (p < 0.001) as compared to placebo. Daytime sleep scores decreased in the EnXtra® group however change was not significant. Sleep quality remained undisturbed in all three arms.Conclusion: The findings demonstrated the psychostimulant efficacy of EnXtra® with no safety concerns on long-term usage.

Action mechanisms and interaction of two key xanthine oxidase inhibitors in galangal: Combination of in vitro and in silico molecular docking studies.

Galangal extract (GE)-based hypouricemic functional food is under-developed due to ambiguous quality control standard that is closely associated with action mechanisms and interaction of key xanthine oxidase (XO) inhibitors (kaempferide and galangin) in GE. In terms of kinetics analysis, fluorescence quenching and molecular docking, kaempferide and galangin showed similar docking posture to xanthine in molybdopterin center, and formed flavonol-XO complexes driven by hydrogen bonding, hydrophobic interaction and van der Waals force, competitively inhibiting XO. Kaempferide, had stronger binding affinity for XO and three more hydrogen bonds with XO than galangin, interacting with critical amino acid residues (Arg880 and Glu802) in catalysis reaction of XO and showing stronger XO inhibitory activity than galangin. The combination of kaempferide and galangin enhanced their binding affinities for XO, showing synergistic inhibition on XO at optimal molar ratio 1:4 that could be quality control standard for GE. This study provided new insights into structure-XO inhibitory activity relationship of methoxylated flavonoids and quality control standard for GE-based hypouricemic functional food.

A combination extract of kaffir lime, galangal, and lemongrass maintains blood lipid profiles, hepatocytes, and liver mitochondria in rats with nonalcoholic steatohepatitis.

Kaffir lime leaves and the rhizomes of galangal and lemongrass are the main ingredients in several Thai foods with desirable medicinal effects. Based on their beneficial activities, this study aimed to indicate the chemical properties and in vivo efficacy of a combination of the herbs at a 1:2:1 ratio in a water extract form. Its volatile constituents were analyzed by gas chromatography coupled with mass spectrometer, which mainly consists of eucalyptol, citronellal, and citral. Clinicohistopathological and electron microscopic studies demonstrated that the extract corrected blood cholesterol, LDL, HDL, and triglyceride levels similarly as simvastatin treatment in association with its antioxidative properties, as indicated by the levels of superoxide dismutase and malondialdehyde in serum and the increment of nuclear factor erythroid 2-related factor 2 levels in hepatocytes. Hepatitis was significantly less severe in rats fed the extract for 14 days than in simvastatin-treated rats. Regarding its immunomodulatory properties, the extract also accelerated hepatic resolution from steatohepatitis during hypercholesterolemia as indicated by the upregulation of vimentin, cytokeratin, and CD206+. Interestingly, liver mitochondria were also preserved in hypercholesterolemic rats treated with the extract in relation to their architecture and the expression of haloacid dehalogenase-like hydrolase domain-containing protein 3 as well as metabolic and energy regulators. Therefore, the study concluded that the water extract of kaffir lime leaves and the rhizomes of galangal and lemongrass has beneficial effects on blood cholesterol, the severity of steatohepatitis, and the maintenance of mitochondrial architecture via its antioxidative and immunomodulatory activities.

Development of galangal essential oil-based microemulsion gel for transdermal delivery of flurbiprofen: simultaneous permeability evaluation of flurbiprofen and 1,8-cineole.

Flurbiprofen (FP) is one of the most potent nonsteroidal anti-inflammatory drugs with very low bioavailability of approximately 12% following transdermal administration, compared to that after oral administration. This study aimed to deliver FP as a microemulsion (ME) gel by transdermal administration. Galangal essential oil (GEO) was extracted from Rhizoma Alpiniae Officinarum and identified by GC-MS. The most abundant constituent was determined to be 1,8-cineole (52.06%). Compared to azone, GEO was proved to exert significantly higher (p < .01) penetration enhancement effect and significantly (p < .001) lower skin cell toxicity. The formulation (FP-GEO-ME gel) was prepared using GEO as an oil phase and a penetration enhancer. Compared to that of FP solution, the enhancement ratio (ER) of FP-GEO-ME gel was 4.06. In addition, more than 25% 1,8-cineole permeated through the rat skin. In vivo pharmacokinetic studies revealed that the AUC0-t of FP after transdermal administration of FP-GEO-ME gel was higher by approximately 4.56-fold than that of marketed FP cataplasms. The relative bioavailability of FP and 1,8-cineole after transdermal administration compared to oral administration of FP-GEO-ME were determined to be 96.58% and 85.49%, respectively. FP-GEO-ME gel significantly inhibited carrageenan-induced hind-paw edema and decreased PGE2 levels in rat serum. GEO-ME gel also exhibited significant anti-inflammatory effects at 2 h after the therapy (p < .05). The synergistic effects of FP and GEO were expected for the application of FP-GEO-ME gel. In conclusion, GEO-ME gel may be a promising formulation for transdermal administration of anti-inflammatory hydrophobic drugs, such as FP.

Discrimination of Two Cultivars of Alpinia Officinarum Hance Using an Electronic Nose and Gas Chromatography-Mass Spectrometry Coupled with Chemometrics.

Background: Alpinia officinarum Hance is both an herbal medicine and a condiment, and generally has different cultivars such as Zhutou galangal and Fengwo galangal. The appearance of these A. officinarum cultivars is similar, but their chemical composition and quality are different. It is therefore important to discriminate between different A. officinarum plants to ensure the consistency of the efficacy of the medicine. Therefore, we used an electronic nose (E-nose) to explore the differences in odor information between the two cultivars for fast and robust discrimination. Methods: Odor and volatile components of all A. officinarum samples were detected by the E-nose and gas chromatography-mass spectrometry (GC-MS), respectively. The E-nose sensors and GC-MS data were analyzed respectively by principal component analysis (PCA), the correlation between E-nose sensors and GC-MS data were analyzed by partial least squares (PLS). Results: It was found that Zhutou galangal and Fengwo galangal can be discriminated by combining the E-nose with PCA, and the E-nose sensors S2, S6, S7, S9 were important sensors for distinguishing different cultivars of A. officinarum. A total of 56 volatile components of A. officinarum were identified by the GC-MS analysis, and the composition and content of the volatile components from the two different A. officinarum cultivars were different, in particular the relative contents of 1,8-cineole and α-farnesene. The classification result by PCA analysis based on GC-MS data was consistent with the E-nose results. The PLS analysis demonstrated that the volatile terpene, alcohol and ester components primarily interacted with the sensors S2 and S7, indicating that particular E-nose sensors were highly correlated with some aroma constituents. Conclusions: Combined with advanced chemometrics, the E-nose detection technology can discriminate two cultivars of A. officinarum, with GC-MS providing support to determine the material basis of the E-nose sensors' response.

Effect of Alpinia officinarum Hance rhizome extract on spermatogram factors in men with idiopathic infertility: A prospective double-blinded randomised clinical trial.

Despite scientific advances, many of the treatments in male infertility remained indeterminate. In recent years, the attention to herbal remedies as an effective treatment for male infertility is considerable. We designed this study to determine the effects of Alpinia officinarum on the results of semen analysis in men with idiopathic infertility. In this clinical trial, seventy-six participants with idiopathic infertility were included in the intervention (plant treatment: n = 31; placebo: n = 29). Participants were randomised to take capsules containing dried extract of A. officinarum rhizome or placebo on a daily (total daily dosage of 300 mg) basis for 3 months. After 12 weeks of intervention, the sperm count and total number of spermatozoa with normal morphology were increased in participants treated with A. officinarum extract compared with the placebo group. The mean sperm count was initially 52 × 106  ± 24 × 106 /ml which changed to 71 × 106  ± 23 × 106 /ml, after intervention (p = 0.043). Also, the mean percentage of spermatozoa with normal morphology was 14.34% ± 9.16% before the treatment which significantly increased to 19% ± 14.89% (p < 0.001). Alpinia officinarum, a traditional medicine remedy, can be effective in the improvement of sperm morphology and sperm count in idiopathic infertility without causing adverse effects.

[Extraction kinetics of volatile oil from galangal by steam distillation].

To reveal the extraction regularity of volatile oil from galangal by GC-MS analysis. The volatile oil in galangal was extracted by steam distillation. The extract was collected every 30 min, the oil part and the water part were separated. GC-MS was used to analyze the extraction liquid collected at different time periods. A total of 140 volatile components were obtained by GC-MS analysis. Among them, the main components were eucalyptus oil alcohol, alpha-pine oil alcohol and 4-terpene alcohol; 22 special components were dissolved in water, 77 special components were dissolved in oil and 41 components were dissolved in both oil and water. With the increase of specific components in water, the content of Eucalyptus in water increased in a linear manner. The increase of eucalyptus oil further promoted the dissolution or dispersion of alpha PN in water, and the change of specific components in oil was positively correlated with the content of Eucalyptus and alpha-terpilenol in oil. The results of principal component analysis show that the physical and chemical properties of the compounds were important factors affecting the distribution of components. PC1 (molecular weight, melting point, boiling point positive correlation), PC2 (negative correlation of refractive index) and PC3 (positive correlation of water solubility) were the main components that lead to the differences in composition distribution. The process of extracting volatile oil from galangal through steam distillation was affected by the physical and chemical properties of volatile components. Some components were specifically distributed in the fragrance and volatile oil system. The endemic components of aromatic water increased the content of the main components in the water system, which may lead to the "emulsification", reduction of the yield and low quality of the volatile oil.

Bioassay-guided isolation of active substances from Semen Torreyae identifies two new anthelmintic compounds with novel mechanism of action.

ETHNOPHARMACOLOGICAL RELEVANCE: Semen Torreyae, the seeds of Torreya grandis Fortune ex Lindley (Cephalotaxaceae) is a well-known traditional Chinese medicinal plant recorded in the Chinese Pharmacopeia (2010 version). It is widely used for treating intestinal parasites in China, owing to its desirable efficacy and safety. However, the anthelmintic compounds in Semen Torreyae have not yet been identified. AIM OF THE STUDY: This study aims to identify the compounds active against helminths from Semen Torreyae. In addition, we tested whether C. elegans strains resistant to currently-used anthelmintic drugs showed cross-resistance to these compounds. METHODS: A bioassay-guided isolation of anthelmintic compounds from Semen Torreyae was performed using a Caenorhabditis elegans (C. elegans) testing model. The structures of active compounds were elucidated by a combination of GC-MS, high resolution MS, and NMR. The median-effect method was employed to generate a combination index (CI) to evaluate the synergistic effect of the anthelmintic compounds. A panel of C. elegans mutant strains resistant against the major anthelmintic drug classes was used to study the cross-resistance to currently-used anthelmintic drugs. A panel of transient receptor potential (TRP) channel mutant strains was also tested to explore the possible mechanisms of action of the anthelmintic compounds. RESULTS: The bioassay-guided isolation led to two active compounds, i.e. galangal acetate (IC50: 58.5 ±â€¯8.9 µM) and miogadial (IC50: 25.1 ±â€¯5.4 µM). The combination of galangal acetate and miogadial resulted in a synergistic effect at IC50, IC70, and IC90 levels (CIs < 1). Galangal acetate and miogadial demonstrated similar activity against drug-resistant C. elegans strains compared to the wild-type strain. In addition, none of the TRP mutants was significantly resistant to galangal acetate or miogadial compared to wild type worms. CONCLUSIONS: We identified the bioactive compounds from Semen Torreyae responsible for its anthelmintic activity: galangal acetate and miogadial. The two anthelmintic compounds demonstrated a synergistic effect against C. elegans. Galangal acetate and miogadial are unlikely to act on the targets of currently-used anthelmintics (ivermectin, levamisole, benomyl and aldicarb), and an action on TRP channels appears to be ruled out as well. In summary, galangal acetate and miogadial are promising anthelmintic hits worth further investigation.

Phytochemical profiles, and antimicrobial and antioxidant activities of greater galangal [Alpinia galanga (Linn.) Swartz.] flowers.

Previous studies of galangal (Alpinia galanga) have focussed mostly on rhizomes but seldom on flowers. A comprehensive look at galangal flowers could reveal additional benefits. The chemical composition of galangal flowers was significantly (p < 0.05) different from that of galangal rhizome. Pentadecane and α-humulene were identified as major compounds of galangal flower essential oil. 1'-acetoxyeugenol acetate was identified as the highest compound in flowers and exhibited the strongest antimicrobial activity among all fractions, with MIC50 values of 34 µg/ml against Staphylococcus aureus and 68 µg/ml against Listeria monocytogenes. Galangal flowers had a 3-fold higher total phenols content than had rhizomes (10.5 vs. 3.33 mg GAE/g powder). The antioxidant activities of different flower fractions varied from 2 to 4.45 mmol  trolox/g phenolics. These findings suggest that antimicrobial and antioxidant agents extracted from galangal flowers could potentially be utilized as natural food preservatives or as therapeutic agents.