Reflux Extract Research Articles

Antioxidant activity and Cytotoxicity against the Cervical Epithelial Carcinoma (HeLa) Cell Line of Crude Ganoderma lucidum mycelial extracts

This study aimed to investigate the total polysaccharide content, antioxidant activity and cytotoxicity on the Cervical Epithelial Carcinoma (HeLa) cells of the ethanol and water reflux extracts from Ganoderma lucidum mycelia. The phenol-sulfuric acid method was used for the standard method of total polysaccharide analysis, the antioxidant was evaluated by DPPH scavenging assay, ABTS assay, and FRAP assay. The MTT assay was used to determine the cytotoxicity of the extracts. The results showed that the contents of total polysaccharide of the ethanol and water reflux extracts were 8.34 ± 0.47 and 207.81 ± 2.45 mg glucose g extract–1, respectively. The assays of antioxidant capacity against DPPH and ABTS demonstrated that the water reflux extract showed a strong radical scavenging effect than the ethanol extract. For the reducing capability of both extracts with FRAP method, they showed the reducing antioxidant activity and possessed concentration dependence by which the water reflux extract had a higher FRAP antioxidant activity than the ethanol extract. The cytotoxic assay of the extracts on normal mammalian cells (Vero cells) revealed that the ethanol extract of Ganoderma lucidum mycelia had a cytotoxic effect on normal mammalian cells at 62.5 – 1,000 mg mL–1, whereas the water reflux extract had no cytotoxicity to normal mammalian cells in all tested concentration. Then, the water reflux extract was selected to determine the cytotoxicity against the HeLa cancer cell line. The results indicated that the water reflux extract of Ganoderma lucidum mycelia exerted a significant cytotoxic effect on HeLa cancer cell line in possessed concentration dependence. The water reflux extract caused a 56.30% decrease in cervical epithelial carcinoma cell viability with concentration of 1 mg mL–1. Therefore, the results of this research demonstrated that the extract of Ganoderma lucidum mycelia provide both antioxidant and anti-cancer activities, which could be used as ingredients pharmaceutical products and functional foods. GRAPHICAL ABSTRACT HIGHLIGHTS Antioxidant activity and Cytotoxicity of Crude Ganoderma lucidum mycelial extracts

Anti-obesity and antihyperlipidemic effects of Phaleria macrocarpa fruit liquid CO2 extract: In vitro, in silico and in vivo approaches

ObjectivePhaleria macrocarpa fruit has been reported to be effectively used in Malaysia and neighboring countries to prevent obesity. Despite its anti-obesity potential, no research has ever predicted the compound-lipase, compound-HMG-CoA reductase interactions and in vivo investigations to further confirm its anti-obesity properties. Thus, the purpose of this study was to assess the anti-obesity and antihyperlipidemic effects of P. macrocarpa by in vitro, computational (in silico), and in vivo assays. MethodInitially, fruit was extracted through liquid CO2 and heating under reflux extraction methods to obtain liquid CO2 extracts (LCE-1 and LCE-2) and heating under reflux extract (HRE), respectively. Subsequently, all three extracts were assessed for antioxidant potentials through in vitro bioassays and FTIR analysis to identify different types of functional groups present in the existing bioactive compounds. Successively, the most active fraction (LCE-2) was subjected to GCMS analysis for the identification of lipid lowering and lipase inhibitors through molecular docking approach. Finally, anti-obesity and lipid lowering effects were further confirmed through an in vivo assay using mice. ResultLCE-2 exhibited higher DPPH (IC50 = 0.172 mg/mL) and FRAP (78.98 AAEmg/g) antioxidant activities and showed more potent peaks in FTIR chromatograms than LCE-1 and HRE. Among the identified tentative bioactive compounds viz. methyl palmitate, palmitic acid, ethyl palmitate, methyl oleate, oleic acid, cis-vaccenic acid, 3-deoxyestradiol and phenol, 2,2′-methylenebis[6-(1,1-dimethylethyl)-4-methyl- were found as an anti-obesity and lipid lowering compounds whose protein–ligand interaction was confirmed by binding affinity, amino acid residues and bonding interactions. Similarly, anti-obesity and lipid lowering findings were also found in a mice model after 6 weeks treatment at a dose of 250 mg/kg b.w. ConclusionBased on the aforementioned in vitro, in silico and in vivo findings, it is concluded that the LCE-2 possesses lipase and HMG-CoA reductase inhibitors that can assist to develop this plant’s extract as an alternative safe lipid lowering herbal medicine in future.

THE EFFECT OF SOLVENT TOWARDS ANTIOXIDANT ACTIVITY OF Vernonia amygdalina Delile LEAVES

Antioxidants guard against reactive oxygen free radicals. Antioxidants neutralize free radicals by adding electrons to them, preventing bodily harm. African leaves (Vernonia amygdalina Delile.) contain an antioxidant. African leaves are the leaves of a thousand diseases because they have pharmacological effects on various diseases. African leaves contain steroids/triterpenoids, glycosides, saponins, tannins, and flavonoids. This study examined Vernonia amygdalina Delile. leaf combination ethanol extract antioxidant properties. The extracts were prepared with the reflux method using methanol, butanol, isopropanol, ethyl acetate, chloroform, and n-hexane. Antioxidant activities were determined with DPPH, ABTS, FRAP, CUPRAC, O-Phenanthroline, Hydroxyl radical scavenging, total phenolics, and total flavonoids method. The data were analyzed using the Principle Component Analysis (PCA) multivariate exploration technique, which was analyzed using the Minitab software. The highest antioxidant activity was found in African leaf reflux extract with methanol solvent by DPPH method (43.8968 ± 0.021), followed by ABTS (73.6223 ± 0.091), FRAP (89.2924 ± 0.087), CUPRAC (13.4745 ± 0.042), O-Phenantroline (99.0977 ± 0.30), and Hydroxyl radical scavenging (33.5703 ± 0.071). The findings show that Vernonia amygdalina Delile. extract exhibit antioxidant activity. According to the findings, Vernonia amygdalina Delile leaf extract with methanol solvent provided the maximum antioxidant activity.

In Vitro, In Silico and Network Pharmacology Mechanistic Approach to Investigate the α-Glucosidase Inhibitors Identified by Q-ToF-LCMS from Phaleria macrocarpa Fruit Subcritical CO2 Extract.

The fruit of Phaleria macrocarpa have been traditionally used as an antidiabetic remedy in Malaysia and neighbouring countries. Despite its potential for diabetes treatment, no scientific study has ever been conducted to predict the inhibitor interaction of the protein α-glucosidase identified in an extract prepared with a non-conventional extraction technique. Hence, the major aim of this research was to evaluate the in vitro antioxidant, the α-glucosidase inhibitors, and the molecular dynamic simulations of the α-glucosidase inhibitors identified by Quadrupole Time-of-Flight Liquid Chromatography Mass Spectrometry (Q-ToF-LCMS) analysis. Initially, dry fruit were processed using non-conventional and conventional extraction methods to obtain subcritical carbon dioxide extracts (SCE-1 and SCE-2) and heating under reflux extract (HRE), respectively. Subsequently, all extracts were evaluated for their in vitro antioxidative and α-glucosidase inhibitory potentials. Subsequently, the most bioactive extract (SCE-2) was subjected to Q-ToF-LCMS analysis to confirm the presence of α-glucosidase inhibitors, which were then analysed through molecular dynamic simulations and network pharmacology approaches to confirm their possible mechanism of action. The highest inhibitory effects of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and α-glucosidase on SCE-2 was found as 75.36 ± 0.82% and 81.79 ± 0.82%, respectively, compared to the SCE-1 and HRE samples. The Q-ToF-LCMS analysis tentatively identified 14 potent α-glucosidase inhibitors. Finally, five identified compounds, viz., lupenone, swertianolin, m-coumaric acid, pantothenic acid, and 8-C-glucopyranosyleriodictylol displayed significant stability, compactness, stronger protein-ligand interaction up to 100 ns further confirming their potential as α-glucosidase inhibitors. Consequently, it was concluded that the SCE-2 possesses a strong α-glucosidase inhibitory effect due to the presence of these compounds. The findings of this study might prove useful to develop these compounds as alternative safe α-glucosidase inhibitors to manage diabetes more effectively.

Absolute Quantification of Isoflavones in the Flowers of Pueraria lobata by qHNMR.

Pueraria lobata (Willd.) Ohwi. is a widely used medicinal plant in Korea, China, and Japan. The flower of P. lobata (Puerariae Flos) contains various bioactive substances such as triterpenoidal saponins and isoflavonoids. In this study, we developed a quantitative analysis of the isoflavones of Puerariae Flos by quantitative proton nuclear magnetic resonance (qHNMR) spectroscopy using the internal calibrant (IC). From the qHNMR results, the isoflavone content was found to be 7.99% and 10.57% for the MeOH sonication extract (PLs) and the MeOH reflux extract (PLr) of Puerariae Flos, respectively. The quantified isoflavone content was validated using the conventional analytical method, high-performance liquid chromatography with ultraviolet detection (HPLC-UV). The present study shows that validated qHNMR spectroscopy is a reliable method for quantifying and standardizing the isoflavone content in Puerariae Flos.

Potency of moringa (Moringa oleifera L.) leaves extract containing quercetin as a depigmentation agent inhibiting the tyrosinase enzyme using in-silico and in-vitro assay

Hyperpigmentation is a disorder of facial skin pigments due to an increase in the process of melanogenesis, which can cause a darkening of skin color. A flavonoid compound with potential as a skin-lightening agent is quercetin, commonly found in Moringa oleifera L. leaves. This study aims to determine quercetin’s affinity and molecular mechanism on tyrosinase enzyme target proteins using an in-silico molecular docking method. Docking of quercetin with the tyrosinase enzyme produced a bond energy value of -7.08 kcal/mol. In comparison, the tropolone as a native ligand with the tyrosinase enzyme produced -4.79 kcal/mol. Quercetin has a strong affinity for the tyrosinase enzyme, indicated by the bond energy results from docking. Quercetin extraction from Moringa oleifera L. leaves using three different extraction methods: maceration, soxhlation, and reflux were made. The chromatogram from the TLC-Densitometry method showed the identification result in maceration and soxhlation extract containing quercetin, while reflux extract did not contain quercetin. The highest quercetin was obtained in the maceration method with a level of 21.57% w/w, while the soxhlation received quercetin as much as 18.49% w/w. In-vitro tests were carried out using a spectrophotometric method using a comparison of kojic acid. The in-vitro test found that IC50 from kojic acid was 48.90 µg/mL and IC50 of the extract from moringa leaf maceration of 115.36 µg/mL. Based on this research, quercetin compounds in Moringa oleifera L. leaves from maceration can potentially be skin-lightening agents.

High or low temperature extraction, which is more conducive to Triphala against chronic pharyngitis?

ObjectiveExplore the effects of high-temperature reflux extraction and low-temperature decompressing inner ebullition on Triphala's chemical composition and anti-chronic pharyngitis activity. MethodsThe network pharmacology was used to analyze the material basis, targets and pathways of Triphala for chronic pharyngitis. HPLC were used to compare the fingerprint profile and content of components between the two extracts. The antioxidant and anti-chronic pharyngitis activities of the two extracts were compared by DPPH assay and ammonia induced chronic pharyngitis model in rats. ResultsThe network pharmacology results showed that the active ingredients of Triphala for chronic pharyngitis are epigallocatechin-3-gallate, (+)-catechin, epicatechin, epicatechin gallate, (+)-gallocatechin, quercetin, luteolin, leucodelphinidin and other flavonoids; phenolic acids such as gallic acid and ellagic acid; alkaloids such as ellipticine, cheilanthifoline; hydrolyzed tannins such as corilagin and chebulic acid. The high-temperature reflux extract and the low-temperature decompressing inner ebullition extract have extremely significant differences in the fingerprint profile. Among them, the content of gallic acid, ellagic acid, chebulic acid, catechin, epicatechin, corilagin, quercetin, and epicatechin gallate in the reflux extract is 1.1–5.3 times as much as decompressing inner ebullition extract. The free radical scavenging ability of reflux extract is significantly stronger than that of decompression extract (p < 0.01), and it has a repairing effect on pharyngeal mucosal damage (reducing keratinization or hyperplasia of mucosal epithelium, reducing inflammatory cell infiltration and bleeding), and reducing IL-1β (P＜0.05), IL-6 (p＜0.05), TNF-α overexpression ability is stronger than the decompressing inner ebullition extract. Conclusionsgallic acid, ellagic acid, chebulic acid, catechin, epicatechin, corilagin and epicatechin gallate are the basic aglycones or oligomers of tannin. High temperature reflux extraction can significantly promote the occurrence of the hydrolysis of tannins and significantly increases the content of these components.Therefore, its anti-chronic pharyngitis activity is enhanced. It is suggested that high temperature reflux extraction should be used in the treatment of chronic pharyngitis.

Chemical constituents from the roots of Asarum sieboldii Miq. var. Seoulense Nakai

Fourteen compounds were isolated from the 95% ethanol reflux extract of Asarum sieboldii Miq. var. Seoulense Nakai, including five phenanthrene derivatives (1–5), three isobutyl amides (6–8), three phenylpropanoids (9–11) and three lignins (12–14). The structures of these compounds were identified by spectroscopic methods and by comparison with the reported spectroscopic data. Among them, compounds 6 and 11 were firstly reported from the family Aristolochiaceae, and compounds 3 and 4 were reported for the first time from the genus Asarum. Additionally, compounds 1, 2 and 8 were isolated from A. sieboldii Miq. var. Seoulense Nakai for the first time. These compounds have shown chemical relationships between A. sieboldii Miq. var. Seoulense Nakai and other species of Asarum as well as those found in the genus Aristolochia in the family Aristolochiaceae.

High-Pressure Supercritical CO2 Extracts of Ganoderma lucidum Fruiting Body and Their Anti-hepatoma Effect Associated With the Ras/Raf/MEK/ERK Signaling Pathway.

As a noted medicinal mushroom, Ganoderma lucidum (G. lucidum) has been reported to have a number of pharmacological effects such as anti-tumor and liver protection. Compared with the common ethanol reflux method, supercritical CO2 extraction has obvious advantages in obtaining antitumor extracts from G. lucidum fruiting body such as short extraction time, low temperature and no solvent residue. However, Using high-pressure supercritical CO2 without entrainer to obtain the antitumor extracts from G. lucidum and studying their anti-hepatoma effect have not been reported. In this study, high-pressure supercritical CO2 extracts obtained under 65, 85, and 105 MPa pressure named as G65, G85, G105 respectively and ethanol reflux extract (GLE) were used to investigate their anti-hepatoma activity and the underlying molecular mechanism. The total triterpenoid content of G85 was significantly higher than that of G65 and GLE, but did not differ significantly from that of G105 by UV and high-performance liquid chromatography. GLE, G65, and G85 could inhibit cell proliferation, arrest cell cycle in G2/M phase, and induce apoptosis in two liver cancer cell lines (QGY7703 and SK-Hep1), of which G85 had the strongest effect. The results showed that the potency of their cytotoxicity of the high-pressure supercritical CO2 extracts on human hepatoma carcinoma cells in vitro was consistent with their total triterpenoid content. G85 exhibited significant anti-hepatoma effect with low toxicity In vivo. Further mechanistic investigation revealed that the anti-tumor effect of these extracts was associated with their inhibition of Ras/Raf/MEK/ERK signaling pathway. Our findings suggest that the high-pressure supercritical CO2 extraction of G. lucidum fruiting body can be used to obtain a triterpenoid-rich anti-tumor agent, which may have potential clinical significance for the treatment of human hepatoma.

FORMULASI SEDIAAN SERBUK EFERVESEN DARI EKSTRAK ETANOL ANGKAK (Monascus purpureus) DENGAN METODE FOAM-MAT DRYING

Indonesia has various types of plants that have potential as medicine to overcome health problems in line with the expansion of the traditional medicine industry. One example of plants from the Zingiberaceae family is Hedychium coronarium, which is not widely known by the community despite it has efficacy as an antibacterial which is not inferior to other Zingiberaceae family. The research was conducted to determine whether the Hedychium coronarium rhizome shown to have antibacterial activity against the growth of Staphylococcus aureus and Escherichia coli, knowing antibacterial compound that contained in Hedychium coronarium rhizome, also knowing the most effective extraction method to take the antibacterial compound from Hedychium coronarium rhizome. The gandasuli rhizome was extracted by maceration and reflux with 96% ethanol. Antibacterial activity test carried out by the agar diffusion method, and the analysis of antibacterial compounds with bioautography, and the results are monitored with vanillin-sulfuric acid. Maceration and reflux extract at a concentration of 90% have antibacterial activity with inhibition zones, each for 8 and 10 mm against Escherichia coli as well as 9 and 10 mm against Staphylococcus aureus. Bioautography results show a positive effect on the appearance of vanillinsulfuric acid. Reflux extract has better antibacterial activity, while the antibacterial compound is an essential oil group.

Bioassay-Guided Different Extraction Techniques of Carica papaya (Linn.) Leaves on In Vitro Wound-Healing Activities.

Herbal plants are traditionally utilized to treat various illnesses. They contain phytochemicals that can be extracted using conventional methods such as maceration, soxhlet, and boiling, as well as non-conventional methods including ultrasonic, microwave, and others. Carica papaya leaves have been used for the treatment of dengue, fungal, and bacterial infections as well as an ingredient in anti-aging products. Phytochemicals analysis detected the presence of kaempferol, myricetin, carpaine, pseudocarpaine, dehydrocarpaine I and II, ferulic acid, caffeic acid, chlorogenic acid, β-carotene, lycopene, and anthraquinones glycoside. Conventional preparation by boiling and simple maceration is practical, simple, and safe; however, only polar phytochemicals are extracted. The present study aims to investigate the effects of three different non-conventional extraction techniques (ultrasonic-assisted extraction, reflux, and agitation) on C. papaya phytochemical constituents, the antioxidant capacity, and wound-healing activities. Among the three techniques, the reflux technique produced the highest extraction yield (17.86%) with the presence of saponins, flavonoids, coumarins, alkaloids, and phenolic metabolites. The reflux technique also produced the highest 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging with an IC50 value of 0.236 mg/mL followed by ultrasonic-assisted extraction (UAE) (IC50: 0.377 mg/mL) and agitation (IC50: 0.404 mg/mL). At tested concentrations (3.125 µg/mL to 500 µg/mL), all extracts do not exhibit a cytotoxicity effect on the human skin fibroblast, HSF1184. Interestingly, reflux and UAE were active fibroblast proliferators that support 85% (12.5 µg/mL) and 41% (6.25 µg/mL) better cell growth, respectively. Additionally, during the early 24 h of the scratch assay, the migration rate at 12.5 µg/mL was faster for all extracts with 51.8% (reflux), 49.3% (agitation), and 42.5% (UAE) as compared to control (21.87%). At 48 h, proliferated cells covered 78.7% of the scratch area for reflux extract, 63.1% for UAE, 61% for agitation, and 42.6% for control. Additionally, the collagen synthesis was enhanced for 31.6% and 65% after 24 and 48 h of treatment for reflux. An HPLC-MS/MS-QTOF (quadruple time-of-flight) analysis of reflux identified nine phytochemicals, including carpaine, kaempferol 3-(2G-glucosylrutinoside), kaempferol 3-(2″-rhamnosylgalactoside), 7-rhamnoside, kaempferol 3-rhamnosyl-(1->2)-galactoside-7-rhamnoside, luteolin 7-galactosyl-(1->6)-galactoside, orientin 7-O-rhamnoside, 11-hydroperoxy-12,13-epoxy-9-octadecenoic acid, palmitic amide, and 2-hexaprenyl-6-methoxyphenol. The results suggested that reflux was the best technique as compared to ultrasonic and agitation.

Comparison of Three Extraction Protocols for the Characterization of Caper (Capparis spinosa L.) Leaf Extracts: Evaluation of Phenolic Acids and Flavonoids by Liquid Chromatography – Electrospray Ionization – Tandem Mass Spectrometry (LC–ESI–MS) and the Antioxidant Activity

Three extraction methods, involving maceration, reflux, and ultrasonic extractions, were used to evaluate the phytochemical content and antioxidant activity of leaf extracts from Capparis spinosa L. collected from five localities in South Tunisia. The results showed that the maceration extract contained the highest content of phenolic acids and flavonoid compounds. The maceration extraction method exhibited the highest antioxidant activities using the reducing power assay, the azinobis (3-ethylbenzothiazoline)-6-sulfonic acid (ABTS+), and the 2,2-diphényl-1-picrylhydrazyle (DPPH°) radical scavenging activity. The determination of the phytochemicals in the extracts was carried out using a quadrupole mass spectrometer equipped with an electrospray ionization source that was operated in the negative ionization mode (LC–ESI–MS). The results allowed the detection of seven phenolic acids in the maceration extract and ultrasonic assisted extraction, but only six phenolic acids in the reflux extract. The major identified phenolic acids were quinic acid, gallic acid, and protocatchuic acid. Ten flavonoids were detected, and their concentrations varied according to the extraction method. Catechin was the most abundant flavonoid in all of the extracts. This investigation provides additional support for the use of caper leaves in medicinal and food applications.

Optimization of choline chloride-glycerol based natural deep eutectic solvent for extraction bioactive substances from Cinnamomum burmannii barks and Caesalpinia sappan heartwoods

Indonesian cassia (Cinnamomum burmannii Blume) is commonly used as a condiment. It reportedly contains a number of major phytochemical constituents such as trans-cinnamaldehyde and coumarin. Sappan wood (Caesalpinia sappan) is a native plant of Southeast Asia that contains brazilin, a widely known red pigment. This study aimed to determine the optimal extraction conditions using a choline chloride-glycerol (ChCl-glycerol)-based natural deep eutectic solvent (NADES) to obtain greater trans-cinnamaldehyde and brazilin levels from Indonesian cassia and sappan wood. The powders of Indonesian cassia and sappan wood were extracted using ChCl-glycerol-based NADES varied at three different levels: ratio of ChCl to glycerol, ratio of powder to NADES, and the amount of water in NADES. All variables were designed using the Box-Behnken design of response surface methodology to provide 15 extraction conditions. The extraction was performed using ultrasonication-assisted extraction for 30 and 50 min for Indonesian cassia and sappan wood, respectively. Determination of the active compound contents was performed using a high-performance liquid chromatography system equipped with a UV-VIS detector at λmax = 280 nm. The optimization results revealed that the highest levels of trans-cinnamaldehyde, coumarin, and brazilin in NADES extracts were 1907.32, 1735.68, and 368.67 μg/ml, respectively, whereas the lowest levels of these compounds were 453.59, 616.76, and 74.21 μg/ml, respectively. The maximal levels exceeded those obtained using a conventional extraction method, in which 5000 μg/ml Indonesian cassia reflux extract contained only 108.45 μg/ml trans-cinnamaldehyde. Similarly, 1000 μg/ml sappan wood contained only 124.64 μg/ml brazilin. ChCl-glycerol-based NADES was suitable for extracting active compounds from Indonesian cassia and sappan wood; moreover, this solvent is more effective than organic ethanolic coventional solvent.

UJI ANTIBAKTERI RIMPANG GANDASULI (HEDYCHIUM CORONARIUM) TERHADAP BAKTERI STAPHYLOCOCCUS AUREUS DAN ESCHERICHIA COLI DENGAN PERBANDINGAN METODE EKSTRAKSI

&lt;p&gt;&lt;em&gt;Indonesia has various types of plants those have potential as medicine to overcome health problems in line with the expanding of traditional medicine industry. One example of plants from the Zingiberaceae family is Hedychium coronarium, which is not widely known by the community despite it has efficacy as an antibacterial which is not inferior to other Zingiberaceae family. The research was conducted to determine whether the Hedychium coronarium rhizome shown to have antibacterial activity against the growth of Staphylococcus aureus and Escherichia coli, knowing antibacterial compound that contained in Hedychium coronarium rhizome, also knowing the most effective extraction method to take the antibacterial compound from Hedychium coronarium rhizome. The gandasuli rhizome was extracted by maceration and reflux with 96% ethanol. Antibacterial activity test carried out by the agar diffusion method, and the analysis of antibacterial compounds with bioautography, and the results are monitored with vanillin-sulfuric acid. Maceration and reflux extract at a concentration of 90% has antibacterial activity with inhibition zones, each for 8 and 10 mm against Escherichia coli as well as 9 and 10 mm against Staphylococcus aureus. Bioautography results show a positive effect on the appearance of vanillin-sulfuric acid. Reflux extract has better antibacterial activity, while the antibacterial compound is an essential oil group.&lt;/em&gt;&lt;/p&gt;&lt;p&gt;&lt;strong&gt; &lt;/strong&gt;&lt;strong&gt;&lt;em&gt;Keywords:&lt;/em&gt;&lt;/strong&gt;&lt;strong&gt;&lt;em&gt; &lt;/em&gt;&lt;/strong&gt;&lt;em&gt;Hedychium coronarium, antibacterial, Staphylococcus aureus, Escherichia coli, bioautography&lt;/em&gt;&lt;em&gt;.&lt;/em&gt;&lt;/p&gt;

In Vitro Assay of Ethanolic Heat Reflux Extract of Nicotiana tabacum L. var Virginia Against Nosocomial Bacteria Pathogen

Tobacco plays an important role in international trade as one of the export commodities. Indonesia is one of the good quality export contributors of tobacco leaves in the world. Nevertheless, tobacco is used only as a raw material for the cigarette industries, and the rise on anti-cigarette regulations prompted the exploration of alternative product from tobacco plants. The content of alkaloids, flavonoids, terpenoids and steroids in tobacco leaves were reported in literatures as antibacterial. Therefore, this study proposed in vitro assay of the ethanolic heat reflux extract (EHRE) of Nicotiana tabacum var. Virginia against nosocomial bacteria pathogen ((Pseudomonas aeruginosa (ATCC 27853), Eschericia coli (ATCC 25922), Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 29212)). Kirby-bauer diffusion method was used for this assay. The concentration of the EHRE for Kirby-bauer assay were 20; 40; 60; 80; and 100%. The presence of clear zones on Kirby-bauer test, against the growth of each nosocomial bacteria pathogen show that tobacco extract has antibacterial effect. Statistical analysis result showed that each extract concentration had significant difference value (p <0,05). This study indicated that the content (alkaloids, flavonoids, terpenoids and steroids) of tobacco leaf extracts (N. tabacum) has potential as antibacterial against nosocomial bacteria pathogen. Nevertheless, optimization of tobacco leaf extract to obtain maximum active ingredient still needs to be done. This study is important for further development of the tobacco leaf extract as antibacterial

Antioxidant Activity and Antibacterial Effects on Clinical IsolatedStreptococcus suisandStaphylococcus intermediusof Extracts from Several Parts ofCladogynos orientalisand Their Phytochemical Screenings

The in vitro antioxidant and antibacterial assays against clinically isolated Streptococcus suis and Staphylococcus intermedius of the extracts prepared by decoction and ethanolic reflux of different parts of Chettaphangki (Cladogynos orientalis Zipp. ex Span), including the leaves, roots, and stems, using 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay and disc diffusion method were conducted. Quantitative analysis of total phenolic and total flavonoid contents in the extracts using spectrophotometric methods was also performed. Finally, phytochemical screening by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) was conducted. Leaf ethanolic reflux extract (100 g) contained the highest total phenolic and total flavonoid contents of 7.21 ± 0.28 μg gallic acid equivalent (GAE) and 11.51 ± 2.02 μg rutin equivalent (RE), respectively. Chettaphangki extracts promoted low antioxidant activity with EC50 values in the range of 0.27–0.48 mg/mL. Extracts and fractions from the roots and stems of this plant promoted low to intermediate antibacterial activity against S. intermedius with the inhibition zones between 7 and 14 mm. The chromatographic data suggested that the leaf extracts of C. orientalis contained rutin while the root and stem extracts contained scopoletin and chettaphanin I. Rutin promoted strong antioxidant activity while chettaphanin I showed low antibacterial activity against Staphylococcus intermedius.

Study on the inhibitory effect of &lt;i&gt;Radix Semiaquilegiae&lt;/i&gt; extract on human hepatoma HepG-2 and SMMC-7721 cells

The main objective of this paper was to investigate the extraction process of ethanol extract of Radix Semiaquilegiae, as well as its inhibitory activity on human hepatoma HepG-2 and SMMC-7721 cells, and to compare the inhibitory effects of different concentrations of ethanol extracts against these two hepatoma cells. Ethanol reflux extraction and ultrasound-assisted extraction with ethanol at room temperature were used in the extraction process, and MTT assay was mainly used in the activity experiment to perform in-vitro anti HepG-2 and SMMC-7721 cell activity screening of ethanol extract, and to calculate the cell inhibition rates of the extracts. The results showed that among the two types of extracts, ethanol reflux extract had more superior antitumour activity to that of the ultrasonic extract, but all of the extracts obtained had certain anti-cancer activities, and the anti-proliferative activity increased with the increase of concentration.

한약복합추출물 HT008의 제조방법에 따른 항염증 효능 및 성분함량 비교연구

Objectives : The biological activities and compound contents of herbal medicine vary depending on manufacturing processes. In this study, we compared anti-inflammatory effects and compound contents of three kinds of multi-herbal extract HT008 produced by different manufacturing processes in order to determine chemical and biological equivalence. Methods : HT008 was produced by three different manufacturing methods: 1. Freeze dried extract of Eleutherococcus senticosus, Scutellaria baicalensis and Angelica sinensis (HT008 FD), 2. Spray dried extract of E. senticosus and S. baicalensis combined with reflux extract of A. sinensis (HT008 SD), 3. Spray dried extract of E. senticosus and S. baicalensis combined with supercritial fluid extract of A. sinensis (HT008 SF). Anti-inflammatory effects were evaluated using acetic acid induced pain model and <TEX>${\lambda}$</TEX>-carageenan induced paw edema model. Compound contents were evaluated by HPLC quantitative analysis of standard compounds of HT008, eleutheroside E, baicalin, z-ligustilide. Results : HT008 FD, HT008 SD and HT008 SF significantly decreased acetic acid induced pain index and <TEX>${\lambda}$</TEX>-carrageenan induced paw edema volume compared with that of control group. There was no significant difference in efficacy among the HT008 FD, HT008 SD and HT008 SF. Standard compound contents of HT008 FD, HT008 SD and HT008 SF were quantified within the range of Korean pharmacopoeia or other research. Conclusions : Three different manufacturing methods of multi-herbal extracts have been developed without noticeable difference in the efficacy or compound contents. The results might be used to establish manufacturing process and industrialization of herbal extracts.

Isolation and Quantitation of Amygdalin in Apricot-kernel and Prunus Tomentosa Thunb. by HPLC with Solid-Phase Extraction

Apricot-kernel and Prunus Tomentosa Thunb. are traditional Chinese herb medicines that contain amygdalin as their major effective ingredient. In this report, three methods for the extraction of amygdalin from the medicinal materials are compared: ultrasonic extraction by methanol, Soxhlet extraction by methanol, and reflux extraction by water. The results show that reflux extraction water containing 0.1% citric acid is the best option. The optimal reflux is 2.5 h and water bath temperature is 60 degrees C. The solid-phase extraction method using C18 and multiwalled carbon nanotube as adsorbents is established the pretreatment of reflux extract, and the result shows that the two adsorbents have greater adsorptive capacity for amygdalin and good separation effect. In order to quantitate amygdalin in Apricot-kernel and Prunus Tomentosa Thunb., a reversed-phase high-performance liquid chromatography method using methanol-water (15:85, for 30 min and pure methanol after 30 min) as mobile phase is developed and a good result is obtained.

A search for amino acids in Apollo 11 and 12 Lunar fines

Lunar fines from the Apollo 11 and 12 missions have been analyzed for amino acids and a wide range of other derivatizable organic compounds by a highly sensitive gas-liquid chromatographic method. A minimum of 3 to 5 p.p.b. of each amino acid would have been detected, but there were no indications of the presence of free amino acids in the samples. The N-trifluoroacetyl n-butyl ester/gas-liquid chromatographic method has been shown to give recoveries greater than 80% when samples containing 5 ng of each of seventeen amino acids were taken through the total derivatization and chromatography, thus showing that losses due to adsorptive and substrate-derivative interactions were minimal. The method was also shown capable of detecting other organic classes as higher alcohols, amines, fatty acids, and dicarboxylic acids. The unhydrolyzed water reflux extract (17 h) of the Apollo 11 fines showed that the concentration of each of the free common amino acids were less than 4 ng/g, however, on hydrolysis of the water extract with 6 N HCl, a large peak was eluted on the glycine—serine region of the chromatogram, and two other peaks at 153° (80 ng/g) and 178° (40 ng/g). The large peak could be contamination as a smaller peak was observed at the same retention temperature in the parallel blank; or these peaks could be due to organic precursors which are hydrolyzed as reported by Fox. The Apollo 12 unhydrolyzed water reflux extract (13 h) was also investigated, and none of the free common amino acids were detected above background ( ca.