White Amorphous Powder Research Articles

Exploring the Optimal Extraction Conditions and Antibacterial Effects of Tea Polyphenols

Tea polyphenols are a general term for polyphenols in tea, which are a white amorphous powder that is easily soluble in water. It is well known that it has many functions, such as antioxidant activity, antibacterial activity, inhibitory enzyme activity and other physiological activities. And as a natural compound, tea polyphenols have good environmental friendliness, and as additives, they can help improve the current environmental pollution situation. Tea polyphenols have extensive application value in various industries, but there is currently a lack of research on their antibacterial properties when combined with daily chemical products. Due to the current lack of research in this area, this experiment used shower gel as an example to study the antibacterial effect of daily chemical products which are added tea extract. This article includes two experiments. One is study on the optimal extraction conditions for tea polyphenol, including two variables on temperature and extraction agent concentration. The other one is study on antibacterial effect of tea polyphenol, by combining variables on store temperature and concentration of tea extract solution, a total of 14 experimental conditions are set up, including the extract acting alone, and acting together with diluted shower gel, and the control group (water, diluted shower gel). The experimental study shows that the inherent content of tea polyphenols varies among different tea varieties and tea extracts is with good antibacterial properties. Generally speaking, the extraction effect at 80 degrees Celsius is significantly better than those at 50 degrees Celsius, and the content also changes with the extraction time. And tea extracts extracted with suitable extraction conditions shows the best antibacterial effect when diluted 100 to 200 times. This article aims to find better extraction methods and antibacterial effect, providing a new approach for the industrial application of tea polyphenols as natural bacteriostat.

Extraction, purification, and quantification of hesperidin from the immature Citrus grandis/maxima fruit Nepal cultivar

Abstract. Manikyam HK, Tripathi P, Patil SB, Lamichhane J, Chaitanya MVNL, Patil AR. 2022. Extraction, purification, and quantification of hesperidin from the immature Citrus grandis/maxima fruit Nepal cultivar. Asian J Nat Prod Biochem 20: 21-26. Hesperidin, a flavanone group of flavonoids and a 7-O-rutinoside of hesperitin is abundantly present in citrus fruits. Pomelo or Citrus grandis (L.) Osbeck/ C. maxima (Burm.) Merr. is grown and cultivated in Nepal as seasonal edible fruit. Pomelo has flavanone and other chemical constituents. Flavanones like naringin, hesperidin, neohesperidin, and naringenin are present in the fruits of C. grandis. Many extraction techniques have been reported in the extraction and purification of hesperidin citrus fruits like sweet lemons, grapefruits, mandarins, etc. However, little information has been provided on hesperidin content in Pomelo fruits. In order to estimate the quantity of hesperidin in the Pomelo fruits of Nepal, we have first time made study. The immature Pomelo fruits of size 2-7 cm were collected from the cultivation field and dried completely. The pulverized fruit powder with moisture content less than 15% was subjected to 5% methanolic acetic extraction and Dimethylformamide (DMF) solvent extraction in Soxhlet apparatus by refluxing at 90°C in 1:15 ration in three subsequent extractions of each 1 hour. The concentrates were crystalized at freezing temperatures in methanol and further purified by repeating the crystallization process. The white amorphous powder was subjected to RP-HPLC analysis method, chromatographic conditions 285 nm wavelength, C18 column 4.6 mm x 15 cm x 5 µm, flow rate 1.2 mL/minute with injection volume 10 µL run time of 30 minutes, column temperature 40°C. Hesperidin yields were 0.12 g/100 g with acetic acid methanol with a purity of 87% and 0.17 g/100 g in DMF extraction with a purity of 90% with recoveries of 87% in acetic acid methanol extraction and 90% in DMF extraction when compared with standard 93%. Further extraction does not yield any hesperidin content indicating a total of 0.15-0.17 g/100 g of hesperidin content in the whole immature fruit of C. grandis/maxima. Thus we found DMF and 5% methanolic acetic solvents as preferable solvents to extract hesperidin.

FORMULATION AND EVALUATION OF EMULGEL OF CEPHALEXIN TO ENHANCE TOPICAL DELIVERY

Emulgel dosage formulation of cephalexin which has enhanced solubility release and bioavailability properties with less inter and intra - subject variability would be desirable. Thus, it was aimed to formulate and evaluate emulgels ofthe cephalexin. By the preformulation studies it is observed that cephalexin is a white to off white amorphous powder having no odor and characterstic taste. Five different type formulations (E-1 to E-5) formed using oil phase having fixed amount of Capric/caprylic Triglyceride (CT) and different ration of Cetyl Palmitate (CP) and fixed quantity of oil soluble emulsifiers Cetyl dimethicone copolyol (CDC) and Sorbitan stearate (Span 60) with Aqueous Phase contain Purified water at pH 6.8. Then observed visually that all formulations were clear, there was no phase separation and Stable during Thermo dynamic changes, heating cooling cycle and centrifugation stressed test. Characterization of all cephalexin emulsion formulations were evaluated for Droplet size, Zeta potential and Poly Dispersity Index (PDI) and % drug content, all emulgels showed excellent results droplet size was found between 1.12 to1.97 µm. All five cephalexin emulgel formulations were odorless, washable, homogeneous, stable and free from grittiness and was evaluated under the various parameters, pH of all formulations were observed at 6.8 and viscosity between 40.23 to 76.38 centi Poise and spreaability between 4.326 to 5.341 gm.cm/sec. In-vitro Drug Release of Cephalexin emulgel formulations were studied and found EG-1 (64.75), EG-2 (68.37), EG-3 (51.32), EG-4 (90.32) and EG-5(94.89). Emulgel formulations EG-4 and EG-5 were found excellent on the basis of cumulative percentage of drug release profile. Release data of both selected formulations was fitted into kinetic models. Drug release data were fitted into the zero order, first order, Higuchi and Peppas-Korsemeyer model of drug release kinetics. Emulgel EG-4 and EG-5 both were followed Zero Order Kinetics explained as continuous and steady release of Cephalexin from the formulation. All five formulations were also tested for stability and found formulation EG-3, EG-4 and EG-5 were stable after 30 days against color change, creaming, creaking and phase separation. KEYWORDS: Emulgel, Cephalexin, Topical delivery, Cetyl dimethicone copolyol, Cetyl Palmitate

Inducible nitric oxide synthase downregulation and cellular antioxidant enzyme enhancing potential of oleanolic acid from Isodon wightii (Bentham) H. Hara against lipopolysaccharide-induced liver damage in bagg albino strain C mice

Background: Isodon wightii (Bentham) H. Hara is an aromatic medicinal herb belongs to Lamiaceae which has been considered as a prolific source of diterpenoids with diverse structural and medicinal values. Objective: The objective of the study is to evaluate in vivo anti-inflammatory activity of oleanolic acid from I. wightii against lipopolysaccharide-induced liver inflammation in Bagg albino strain C mice. Materials and Methods: Fractions were obtained using silica gel 60–120 mesh column chromatography, and structural elucidation was done using spectroscopic studies. Hepatoprotection, inducible nitric oxide synthase (iNOS) gene downregulation and the levels of cellular antioxidant enzymes with their respective gene expression were analyzed using reverse transcriptase-polymerase chain reaction (RT-PCR). Results: White amorphous powder (58 mg) was isolated from petroleum ether extract (15 g) eluted with petroleum ether: ethyl acetate mixture (90.5:9.5), and structure has been elucidated as oleanolic acid. 50 μg of oleanolic acid showed effective hepatoprotection and multiple fold increase in the level of antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase. RT-PCR analysis revealed that oleanolic acid had a remarkable effect on iNOS downregulation and antioxidant genes upregulation at 50 μg concentration. Conclusion: The present study proves that oleanolic acid possesses liver protecting activity with a positive effect on increased antioxidant enzymes production. The triterpenoid acid and oleanolic acid could be a suitable natural source for preparing hepatoprotective tonics.

High‐purity amorphous silica from rice husk: Preparation and characterization

AbstractAmorphous silica has a wide range of applications in chemical, biological and environmental sectors. In this work, we studied the preparation of amorphous silica from rice husk, an abundant silica‐rich agricultural waste in Vietnam. We investigated and compared two methods of preparation: (1) via selective removal of alkaline and alkaline‐earth oxides from rice husk ash to yield amorphous silica; and (2) via selective precipitation of silica from aqueous solutions which had been prepared by dissolving rice husk ask in NaOH solutions. The obtained silica was characterized by using XRD, EDX, SEM and BET techniques. The results show that the best obtained silica is a white amorphous and porous powder with high purity of SiO2, in the form of aggregates of 50 nm‐sized primary particles, and a specific surface area of 186 m2/g. The SiO2 recovery is around 8.3 (g) per 100 (g) rice husk. Moreover, in the context of the present work, method (1) can be a more straightforward route for the preparation of amorphous silica in terms of silica's purity, porousity and amorphousness. In contrast, method (2) is shown for a lower grade of silica purity which could be due to strong adsorption of ions from the solution onto the amorphous SiO2 precipitate during precipitation process. The obtained silica has promising applications as adsorbents, fillers and fertilizers.

From 5,000 crude plant and fungi extracts screening to leishmanicidal compounds identification: The challenge goes on

Leishmaniasis are diseases caused by protozoa parasites of the Leishmania genus. About 0.7 million to 1.3 million new cases occur worldwide annually of cutaneous leishmaniasis [1]. Furthermore, the number of cases is increasing globally at an alarming rate. The need for alternative treatments for leishmaniasis has been stimulating bioprospecting programs. Thus, we screened more than 5,000 thousands plant and fungi ethanol extracts (EX) at 20 µg/ml using the amastigote-like forms (ALF) of Leishmania (leishmania) amazonensis (IFLA/BR/67/PH8 strain) [2]. The active extracts were assayed for cytotoxic activity against macrophages (Mθ). Those without cytotoxicity were tested on Mθ infected with the amastigote form of L. amazonensis. Twenty-seven extracts passed the screen and were selected for chemical fractionation aiming at the identification of the leishmanicidal compounds. We hereby describe the investigation of two of them: Baccharis platypoda (B) and Cyrtocymura scorpioides (C), from the Asteraceae family. Both EX from the leaves were subjected to HSCCC. From B, 241 fractions were collected and grouped on 21 fractions according to TLC. These fractions were evaluated against the intracellular parasites. One of the fractions killed 100% of the amastigotes forms at 50 µg/mL and was fractionated (RP-HPLC) to afford a white amorphous powder. This compound killed 72% of the amastigotes at 50 µg/mL (155µM). Analysis of MS and NMR data allowed us to identify a new clerodane diterpene: (1R,3R,4R,8aR)-3-(hydroxymethyl)-4-[(E)-5-hydroxy-3-methyl-pent-3-enyl]-4,8,8a-trimethyl-1,2,3,4a,5,6-hexahydronaphthalen-1-ol. From C, 260 fractions afforded 24 groups after TLC analysis. Group 2 (20 µg/mL) killed 89% of the ALF. RP-HPLC separation afforded chlorogenic acid (CAc) which killed 70% of the ALF (56µM, 20 µg/mL). Amphotericin B was used as control. LC-MS/MS of the other six active fractions indicated the presence of rutin, isoquercitrin, quercitrin.

Chemical Constituents of Euphorbia sororia

Euphorbia sororia Schrenk, an annual herbaceous plant of the genus Euphorbia L. (Euphorbiaceae), is distributed mainly in Central Asia and the Hetian region of Xinjiang in China. Having diuretic, antihypertensive, antiasthmatic, and purgative properties, it is a commonly used medicinal material of Uighur doctors and has been recorded in the pharmacography of Uighur [1]. In this work, we report on the chemical constituents of the aerial parts of E. sororia. The 95% EtOH extracts of the plant were partitioned with EtOAc and n-BuOH, and the corresponding residues, submitted to different chromatographic techniques, such as silica gel, Sephadex LH-20, and MCI-gel columns, led to the isolation and identification of nine chemical compounds. Their structures were identified as astragalin (1), kaempferol-3-O-D-galactoside (2), rutin (3), 24-methylene cycloartanol (4), heptacosanol (5), octacosanol (6), adenosine (7), -sitosterol (8), and daucosterol (9), respectively. Compounds 1, 4–6 are reported from the plant for the first time [2, 3]. All the purified compounds were identified by spectral analysis (1H and 13C NMR, EI-MS) and by comparison with the literature data [2–8]. Plant Material. The plant materials were collected in the Tacheng region of Xinjiang (June 2004) and authenticated by Prof. Zhang Yan Fu (Xinjiang Institute of Materia Medica, China) for E. sororia. Extraction and Isolation. The air-dried aerial parts of E. sororia (10 kg) were extracted three times with 95% aqueous ethanol (v/v) (50 L 3 h) at reflux to give a crude extract (1.8 kg) after concentration at reduced pressure. The extract was suspended in water and extracted with petroleum ether, EtOAc, and n-BuOH successively. The petroleum ether fraction (45 g) was subjected to repeated silica gel column chromatography and eluted with petroleum ether–EtOAc gradient to give compounds 4–7. The EtOAc fraction (45 g) was subjected to repeated silica gel column chromatography and eluted with petroleum ether–EtOAc–MeOH gradient to give compounds 8 and 9. The n-BuOH fraction (30 g) was chromatographed over an AB-8 macroporous absorbent resin column with gradient elution by water, 10% EtOH, and 30% EtOH successively. The 30% EtOH fraction (10 g) was chromatographed over Sephadex LH-20 and MCI gel columns and eluted with H2O–CH3OH in a gradient manner to afford compounds 1–3. Astragalin (1). C21H20O11, light yellow needles (MeOH), mp 163–165 C. HCl-Mg reducion reaction is positive. 1H NMR (600 MHz, CD3OD, , ppm, J/Hz): 8.05 (2H, d, J = 8.4, H-2 , 6 ), 6.88 (2H, d, J = 8.4, H-3 , 5 ), 6.20 (1H, d, J = 1.2, H-6), 6.40 (1H, d, J = 1.2, H-8), 5.35 (1H, d, J = 6.8, H-1 ). 13C NMR (150 MHz, CD3OD, , ppm): 156.6 (C-2), 133.4 (C-3), 177.0 (C-4), 160.1 (C-5), 98.2 (C-6), 164.2 (C-7), 93.7 (C-8), 156.6 (C-9), 104.2 (C-10), 120.9 (C-1 ), 131.0 (C-2 , 6 ), 115.1 (C-3 , 5 ), 160.1 (C-4 ), 101.1 (C-1 ), 74.5 (C-2 ), 76.4 (C-3 ), 71.9 (C-4 ), 76.6 (C-5 ), 60.9 (C-6 ). Kaempferol-3-O-D-galactoside (2). C21H20O11, yellow powder (MeOH), mp 231–232 C. IR (KBr, , cm–1): 3455, 3173, 1656, 1590, 1558, 149, 1302, 1264, 1081, 798. 1H NMR (600MHz, CD3OD, , ppm, J/Hz): 12.50 (H, s, 5-OH), 8.08 (2H, d, J = 9.0, H-6 ), 6.84 (2H, d, J = 9.0, H-3 , 5 ), 6.40 (1H, d, J = 1.2, H-8), 6.20 (1H, d, J = 1.2, H-6), 5.35 (1H, d, J = 7.2, H-1 ). 13C NMR (150 MHz, CD3OD, , ppm, J/Hz): 176.6 (C-4), 164.2 (C-7), 160.1 (C-5), 159.1 (C-4 ), 156.3 (C-9), 156.3 (C-2), 133.2 (C-3), 131.4 (C-2 , 6 ), 120.2 (C-1 ), 114.6 (C-3 , 5 ), 104.4 (C-10), 102.0 (C-1 ), 98.2 (C-6), 93.7 (C-8), 75.7 (C-5 ), 73.9 (C-3 ), 71.7 (C-2 ), 68.0 (C-4 ), 60.1 (C-6 ). 24-Methylene Cycloartanol (4). C41H52O, white amorphous powder (CHCl3), mp 122–123 C. EI-MS m/z 441 [M + H]+. 1H NMR (600 MHz, CDCl3, , ppm, J/Hz): 0.33, 0.55 (each 1H, d, J = 4.2, H2-19), 0.81 (3H, s, H3-30), 0.90 (3H, s, H3-28), 0.90, (3H, d, J = 6.5, H3-21), 0.97 (6H, s, H3-18, 29), 1.02 (3H, d, J = 6.9, H3-27), 1.03 (3H, J = 6.9, H3-26), 4.62, 4.67 (each 1H, br.s, H2-31). 13C NMR (150 MHz, CDCl3, , ppm, J/Hz): 32.0 (C-1), 30.40 (C-2), 78.9 (C-3), 40.5 (C-4), 47.1

Chemical Components of the Tuber of Alocasia cucullata

Alocasia cucullata Schott is mainly distributed in Guangxi, Guangdong, Sichuan, Guizhou, and Fujian Provinces in China (Wu, 1990). Its tuber (Chinese name “jianweiyu”) is a well-known ethnic medicine of the Zhuang nationality in China and is used to treat cancer, reduce swelling, for detoxification, and to ease pain. The antitumor effects of this herb have been preliminarily confirmed because of its widespread clinical application in China for centuries. Nevertheless, up to now, no systematic study has been done on the antitumor constituents of A. cucullata, and the occurrence of an N-acetyl-D-lactosamine (LacNAc)-specific lectin has been reported in a previous investigation of A. cucullata [1]. Our previous pharmacological study further showed the presence of cytotoxic activity in a 50% ethanol extract [2]. In this article we report the isolation of a total of 12 substances: steroid (1), alkaloids (2–4), and phenolic substances (5–12) for the first time from the genus Alocasia. Plant Material. The tubers of A. cucullata were collected in Sichuang Province, China, in May 2011 and identified by Prof. Bao-Kang Huang (Second Military Medical University, Shanghai, China). A voucher specimen (Zheng 5580) has been deposited in the Herbarium of the Shanghai University of Traditional Chinese Medicine. Processing of Plant Material. The tubers of A. cucullata (5 kg) were powdered and extracted three times with 50% EtOH. The extract was concentrated under reduced pressure to give a crude residue (200 g), which was extracted successively with petroleum ether, ethyl acetate, n-butanol. The n-butanol fraction, concentrated to dryness (30 g), was further fractionated on a silica gel chromatography column (600 g of silica gel 60, 70–230 mesh; CHCl3–MeOH–H2O [(15:0.4:1,10:0.4:1, and 5:0.4:1) gradient)] to give four fractions, Frs. A, B, C, and D which were further fractionated on a silica gel chromatographic column (RP-18): CC of Fr. A (2.2 g) (RP-18, 200 g; CH3OH–H2O 4:1) gave 1 (30 mg), 2 (36 mg), 3 (30 mg), and 4 (28 mg); CC of Fr. B (6.2 g) (RP-18, 400 g; CH3OH–H2O 1:1) gave 3 (20 mg), 4 (100 mg), 5 (46 mg), 6 (30 mg), 7 (58 mg), and Fr. B (0.9 g), which were pooled. Fraction B was chromatographed on Sephadex LH-20 (CH3OH–H2O 1:1) yielding 8 (32 mg). CC of Fr. C (5.2 g) (RP-18, 400 g; CH3OH–H2O 2:1) gave 9 (30 mg) and 10 (34 mg). CC of Fr. D (2.1 g) (RP-18, 400 g; CH3OH–H2O 4:1) gave 11 (21 mg) and 12 (25 mg). Compound 1, C35H60O6, white amorphous powder (MeOH), mp 134–135 C. ESI-MS m/z 577 [M + H]+. Its physical constants and spectral data were in agreement with those of -daucosterol [3]. Compound 2, C10H13N5O4, white amorphous powder (MeOH), mp 234.4–235.4 C. ESI-MS m/z 268 [M + H] +. Its physical constants and spectral data were in agreement with those of -adenosine [4]. Compound 3, C6H6N2O, white amorphous powder (MeOH), mp 129–130 C. ESI-MS m/z 121 [M – H]–. Its physical constants and spectral data were in agreement with those of niacinamide [5]. Compound 4, C9H7NO, white gum (MeOH). ESI-MS m/z 146 [M + H] +. Its physical constants and spectral data were in agreement with those of 1H-indole-3-carboxaldehyde [5]. Compound 5, C9H10O3, yellow oil, mp 49–50 C. ESI-MS m/z 167 [M + H]+. Its physical constants and spectral data were in agreement with those of paeonol [6]. Compound 6, C7H6O5, yellow powder. ESI-MS m/z 171 [M + H] +. Its physical constants and spectral data were in agreement with those of gallic acid [6].

Chemical Constituents of Inula helenium

The genus Inula comprises about 100 species, which are widely distributed in Asia, Europe, and predominantly in the Mediterranean region. Among them, more than 20 species are distributed in China. Inula helenium (Compositae) is a widely occurring perennial herb in Europe and East Asia [1]. Its roots have been traditionally used as an expectorant, antitussive, in diaphoresis, and as a bactericidal agent in folk medicine. The roots of I. helenium were collected in August 2004 from the An Guo herbal material market (China). A voucher specimen (No. 2005–06) was deposited in the laboratory of medicinal natural product chemistry, Hebei Medical University. Several kinds of chemical compounds have been isolated from this plant, including sesquiterpene lactones and two new eudesmanes, 1 -hydroxy-11,13-dihydroisoalantolactone and 3 -hydroxy-11,13-dihydroalantolactone [2]. The air-dried roots (3.0 kg) of I. helenium were pulverized and extracted three times with 95% EtOH. The extract (791.2 g) was suspended in brine and partitioned sequentially with petroleum ether, dichloromethane, and ethyl acetate, successively. The dichloromethane extract (232.3 g) was chromatographed on silica gel using CH2Cl2–Me2CO (30:1 1:1) with subsequent preparative TLC and reversed-phase HPLC, leading to compounds 1 and 2. The ethyl acetate extract (88.3 g) was separated by silica gel with EtOAc–MeOH (10:1) followed by stepwise addition of MeOH to afford 15 fractions. Fractions 2 and 3 were purified over Sephadex LH-20 column chromatography to afford compound 3. All compounds were confirmed by 1H NMR and 13C NMR methods. Compound 1, C27H28N2O4, colorless crystals (MeOH), mp 182–184 C, identified as aurentiamide acetate [3–5]. Compound 2, C36H60O6, white amorphous powder (MeOH), mp 92–93 C, characterized as urs-12-en-18 -H-3-O-D-glucopyranoside [6]. Compound 3, C18H14O7, bright yellow amorphous powder (MeOH). Characterized as caffeic acid anhydride [7]. Melting point was not avaliable. Compounds 1–3 were isolated from I. helenium for the first time.

Phenolic compounds from roots of Rhodiola litvinovii

Plants of the genus Rhodiola (Crassulaceae) typically have a high content of biologically active compounds. R. rosea (golden root) is a medicinal plant that is used in folk and modern medicine. Therefore, it seemed interesting to study other species of Rhodiola. Four species of this genus are indigenous to Uzbekistan [1]. The chemical composition of the population of R. litvinovii Boriss. growing in Uzbekistan is practically unstudied [2]. Six phenolic compounds were isolated previously from a population of this plant growing in Siberia (Russia) [3–5]. We studied roots of R. litvinovii collected during flowering in the western Tien-Shen (Pskem Range, Bostandyk District, Tashkent Region, RU) in September 2010. For this, roots of R. litvinovii (2000 g) were extracted exhaustively with EtOH (40%) at room temperature by percolation (24 h). The combined extracts were evaporated in vacuo. The condensed thick residue (0.7 L) was diluted with H2O (1:1) and worked up successively with petroleum ether, CHCl3, EtOAc, and n-BuOH. The condensed EtOAc extract was placed on a chromatography column of silica gel that was eluted sequentially with CHCl3 and CHCl3:MeOH mixtures of various proportions. The resulting separate eluates were re-chromatographed over Sephadex LH-20 with elution by H2O:MeOH mixtures with increasing gradient of MeOH. The chromatographic separation isolated four known compounds that were identified by comparing their IR, UV, PMR, and 13C NMR spectra with the literature. p-Tyrosol (p-hydroxyphenyl-ethanol) (1), white needle-like crystals, C8H10O2, mp 94–95°C (CHCl3:MeOH, 4:1). UV spectrum (MeOH, max, nm): 204.36, 223.93, 278.27. IR spectrum (KBr, , cm –1): 3390–3144 (OH), 2879 (CH2), 1613, 1598, 1513 (aromatic ring), 817 (p-substituted benzene ring). 13C NMR spectrum (100 MHz, CD3OD, , ppm): 131.20 (C-1), 130.99 (C-2), 116.28 (C-3), 156.88 (C-4), 116.28 (C-5), 130.99 (C-6), 39.54 (C-7), 64.72 (C-8) [6, 7]. Salidroside [p-hydroxyphenyl-( -D-glucopyranosyl)-ethanol] (2), white amorphous powder with a yellow tint, C14H20O7, mp 163–164°C (CHCl3:MeOH:H2O, 26:14:3). UV spectrum (MeOH, max, nm): 203.06, 223.90, 277.60. IR spectrum (KBr, , cm–1): 3596-3271 (OH), 2933, 2878 (CH2), 1883, 1615, 1518 (benzene ring), 1241 (phenol hydroxyl), 1073, 1023 (pyranoside), 900 ( -glycoside bond). 13C NMR spectrum (100 MHz, CD3OD, , ppm): 131.06 (C-1), 130.80 (C-2), 116.25 (C-3), 146.51 (C-4), 116.25 (C-5), 130.80 (C-6), 36.50 (C-7), 71.77 (C-8), 104.49 (C-1 ), 72.23 (C-2 ), 78.07 (C-3 ), 71.77 (C-4 ), 78.22 (C-5 ), 62.88 (C-6 ) [8]. Gallic acid (3), crystals, very soluble in alcohol and hot water, C7H6O5, mp 238–240°C. UV spectrum (EtOH, max, nm): 211, 274, Rf 0.65 (BAW, 4:1:2), 0.34 (2% AcOH); gave a blue color with iron chloride solution (1%). Identification of gallic acid (3) was confirmed by an XSA. (–)-Epigallocatechin-3-O-gallate (4), brown amorphous powder, C22H18O11, mp 210–211°C. UV spectrum (MeOH, nm): max 205, 275; min 255. IR spectrum (KBr, , cm –1): 3386, 2929, 1728, 1616, 1527, 1451, 1283, 1206, 1046, 776. 13C NMR spectrum (100 MHz, CD3OD, , ppm): 77.22 (C-2), 68.56 (C-3), 25.42 (C-4), 155.83 (C-5), 95.19 (C-6), 156.48 (C-7), 94.53 (C-8), 156.42 (C-9), 98.08 (C-10), 129.44 (C-1 ), 105.54 (C-2 ), 144.89 (C-3 ), 132.40 (C-4 ), 145.29 (C-5 ), 105.54 (C-6 ), 120.17 (C-1 ), 108.91 (C-2 ), 144.89 (C-3 ), 138.39 (C-4 ), 145.29 (C-5 ), 108.91 (C-6 ), 166.20 (COO) [9, 10]. These compounds were isolated for the first time from this plant species.

Chemical constituents of Paederia scandens

Paederia scandens (Lour.) Merr. is a plant of the family Rubiaceae and genus Paederia. The aerial parts can be used as folk medicine for the treatment of pain and inflammation. The Chinese Pharmacopoeia (1977 edition) listed P. scandens as an official drug. Flavonoids 1 , iridoids 2, 3 , and triterpene saponins 4 were isolated in previous phytochemical studies of the plant. Recently, we have systematically investigated the chemical constituents of the aerial parts. This study was carried out with the purpose of validating the medical use of the aerial parts. In this paper, we described the isolation and structural elucidation of 15 compounds. Among them, compound 1 is a new iridoid compounds, compounds 3 and 4 were isolated from this plant for the first time, and the other compounds are known. The aerial parts of P. scandens were collected at Medicinal Materials Corporation in Sichuan Province, China, in May 2006 and identified by Tong Wu (Shanghai Institute of the Pharmaceutical Industry). A voucher specimen was deposited in Shanghai Institute of the Pharmaceutical Industry, Shanghai, China (collection No. SIPI-060321). The aerial parts of P. scandens (50 kg) were extracted with ethanol (70%), and the ethanol extract was partitioned with petroleum ether, CHCl3, EtOAc, and n-BuOH. Each fraction was purified by column chromatography with silica gel, Rp-18, Sephadex LH-20, and preparative HPLC to yield compounds 1–15. The compounds were investigated by spectroscopic methods, including NMR and mass spectrometry. All compounds were identified as ethyl paederosidate (1), methyl paederosidate (2) 5 , scopoletin (3) 6 , liriodendrin (4) 7 , paederoside (5) 5 , paederosidic acid (6) 8 , deacetyl-daphylloside (7) 9 , dimer of methyl paederosidate and paederosidic acid (8) 2 , paederoscandoside (9) 10 , saprosmoside E (10) 11 , cleomiscosin D (11) 12 , daphylloside (12) 13 , 3-hydroxy4-methoxybenzaldehyde (13) 14 , O-hydroxylbenzoic acid (14) 15 , and 2,5-dihydroxybenzoic acid methyl ester (15) 16 . Compound 1 was obtained as a white amorphous powder with [ ]23 D +11.52 (c 0.525, MeOH). Its molecular formula, C20H28O12S, was established by ESI-MS combined with HR-ESI-MS (m/z 515.1201 [M + Na]+, C20H28O12SNa, calcd 515.1199). As shown in Table 1, the 1H NMR spectrum of 1 indicates the presence of two protons [ 7.64, (d, J = 1.1, H-3) and 6.02, (d, J = 1.4, H-7)], one ethoxy [4.20 (m, J = 7.1, 11-OCH2), 1.29 (t, J = 7.1, 15-CH3)], one sulfur methyl group (2.34, s), and seven protons from the sugar moiety. The 13C NMR spectrum of 1 exhibits signals of 20 C-atoms, which are similar to 19 C-atoms in compound 2 that have 10 carbons of the main iridoid skeleton 17 and six sugar carbons, including one ester carbonyl ( 173.17), one methyl sulfur (13.84), and one ethoxy carbon (61.57 and 14.88). The presence and position of sulfur atom were determined by HR-ESI-MS and comparison of the NMR data with paederoside 8 corresponding to methyl thiocarbonate group. The HMBC data of 1 show an ester carbonyl (C-12) correlated with the sulfur methyl group and H-10, indicating the presence of the partial structure of two nonequivalent H in CH3–S–COO–CH2–. The cross peaks between 11-OCH2 and C-11 and between an anomeric carbon signal [ 100.97, (C-1 )] of the glucopyranose and H-1 [5.06, (d, J = 9.0 Hz)] of the aglycone detected in the HMBC spectrum clearly indicate the presence of an ester functional group and the glycosidic linkage of the glucopyranosyl moiety to the C-1 position of aglycone, respectively 18, 19 . Similarities were found in the 1H and 13C NMR spectra between compound 1 and compound 2, the well-known compound methyl paederosidate 2, 3 which was also isolated in this study. Similarities were also found in the HMBC, HMQC, and NMR spectra between compounds 1 and 2, showing the optical rotation of compound 1 of [ ]23 D +11.52 (c 0.525, 589 nm, MeOH) and of methyl paederosidate of [ ]20 D +12.4 (c 0.90, MeOH). Therefore, compound 1 should have similar absolute configuration to the iridoid glucoside compound 2. Thus, compound 1 was identified as a novel iridoid glucoside, namely ethyl paederosidate.

Chemical Constituents of the Ficus elastica Leaves and Their Antioxidant Activities

Ficus elastica Roxb. Ex Hornem also known as rubber tree, is a tree species belonging to the Moraceae family. This plant grows profusely without any agronomic management and survives well under extreme environmental conditions such as high temperature and limited water supply. F. elastica possesses antimicrobial activity and the leaves extract is used for the treatment of skin infections and skin allergies, as well as a diuretic agent. In addition, several chemical constituents from F. elastica have been investigated. 1 Previously, we reported the antioxidant activities of compounds from the leaves of F. microcarpa and F. callosa . 2,3 In our screening project for antioxidant activities from Ficus species, the leaves of F. elastica was found to exhibit significant antioxidant activities. As part of our continuing research into the biological activities of F. elastica leaves, we report herein the isolation, structural elucidation, and evaluation of antioxidant activities of extracts, as well as two new compounds, ficuselastic acid (2) and (1'S,6'R)-8-O-β-Dglucopyranosyl abscisate sodium (7), along with twelve known compounds from the leaves of F. elastica. Compound 2 was obtained as a white amorphous powder and its molecular formula was determined to be C14H16O6 by ESI-MS at m/z 279 [M–H] – (negative) and HR-ESI-MS at m/z 279.0858 (Calcd C14H15O6 for 279.0874). The 1 H-NMR spectrum of 2 showed signals for a secondary methyl at δH 1.23 (d, 7.0), an acetyl methyl at δH 2.04 (s), an oxygenated proton at δH 5.19 (m), and an olefinic proton at δH 6.19 as listed in Table 1. The 13 C-NMR and DEPT spectra of 2 revealed seven non-protonated carbons, consistent with two carbonyls (δC 172.7 and 174.9) and five olefinics (δC 105.2, 122.3, 139.1, 161.9, and 164.9), together with seven pro

Benzyl Glycosides from the Aerial Parts of Gynostemma laxum and Their NF-κB Inhibitory Activity in HepG2 Cells

Nuclear transcription factor-κB (NF-κB) represents a family of Rel domain-containing proteins including RelA, RelB, c-Rel, NF-κB1, and NF-κB2. The activation of NF-κB has been linked to multiple pathophysiological conditions such as cancer, arthritis, asthma, inflammatory bowel disease, and other inflammatory conditions. NF-κB can be activated by various stimuli, such as microbial and viral products, cytokines, DNA damage and noxious chemicals. The induction of a number of pro-inflammatory mediators occurs as the results of increased inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) activities. Therefore, suppression of iNOS and COX-2 activities can be an important approach to preventing inflammation in organs. NF-κB and the signaling pathways that regulate many physiological processes, including the innateand adaptiveimmune responses, cell death, and inflammation have become a focal point for intense drug discovery and development efforts. Indeed, growing evidences has validated NF-κB as a target for anti-inflammatory and anticancer agents. Gynostemma genus (Cucurbitaceae) is a climbing vine, attaching itself to supports using tendrils. Over 30 species of Gynostemma are known to grow throughout China, mostly in the South-west. In Vietnam, there are only two species of the Gynostemma genus recorded to date, Gynostemma pentaphyllum (Thunb.) Makino and Gynostemma laxum (Wall.) Cogn. Of these, G. laxum is distributed throughout Vietnam from the plains to mountainous areas. The aerial parts of G. laxum have been used as a bitter stomachic, a laxative, anti-inflammatory as well as anti-obesity. Up to date, no report has been reported about chemical components from this plant. In our screening project for antiinflammatory agents from natural sources, we found G. laxum to possess anti-inflammatory effects. As a part of our continuing research to elucidate the anti-inflammatory compounds, we report herein the isolation, structural elucidation, and evaluation of the inhibitory effects on NFκB activation and iNOS and COX-2 expression of a new benzyl glycoside, 2,4-dihydroxybenzyl-O-α-L-rhamnopyranoside (1), along with nine known compounds from G. laxum. The dried aerial parts of G. laxum were extracted with MeOH and partitioned with n-hexane and ethyl acetate, and water. From these fractions and subsequently separation, one new and nine known compounds were isolated (see Figure 1). Compound 1 was obtained as a white amorphous powder and its molecular formula, C13H18O7, determined on the basis of ESI MS at m/z 287 [M+H] (positive) and HR ESI MS at m/z 287.1147 [M+H] (calcd C13H19O7 for 287.1131). The H-NMR spectrum of 1 (DMSO-d6) (Table 1) showed the following signals: one secondary methyl group at δH 1.03 (d, J = 6.0 Hz), assigned to H-6 of the rhamnose; one anomeric proton at δH 4.72 (br s); one oxygenated methylene (δH 4.42 and 4.50); and one 1,2,4-trisubstituted aromatic ring with ABX coupling patterns [δH 7.20 (d, J = 8.0 Hz), 7.25 (d, J = 8.0 Hz), and 7.96 (s)]. The C-NMR and DEPT spectra (Table 1) revealed 13 carbon signals, of which, 7 signals were assigned to a benzyl moiety and 6 signals belonged to rhamnose moiety. The Hand C-NMR data of 1 (Table 1) were similar to those of benzyl-O-α-L-rhamnopyranoside except for an appearance of two hydroxyl groups at aromatic ring. In the HMBC spectrum (see Figure 2), two proton signals at δH 4.45 and 4.50 of oxygenated methylene group correlated with carbons C-1 (δC 126.3), C-2 (δC 148.1), and C-6 (δC 126.4); one proton signal at δH 7.25 (H-6) correlated with carbons C-2 (δC 148.1), C-4 (δC 154.0), and C-7 (δC

Triterpenes of Euonymus alatus and their cytotoxic activity

observed in the mass spectra showed the molecular formula to be C 30 H 50 O 2 . The 1 H NMR spectrum revealed the presence of seven methyl groups: H 0.86 (3H, d, J = 6.7 Hz, H-23), 0.72 (3H, s, H-24), 0.87 (3H, s, H-25), 1.00 (3H, s, H-26), 1.07 (3H, s, H-27), 1.11 (3H, s, H-28); one of them was attached to a tertiary methyl, while the other six were attached to a quaternary carbon. The signal of the proton connected with the hydroxyl group carbon consisted of two parts: 3.34 (1H, d, J = 10.7 Hz) and 3.42 (1H, d, J = 10.7 Hz), while the signal of the secondary methyl carbon connected with the hydroxyl group appeared at C 71.9, showing that this hydroxyl group was attached to C-30 [11]. The 1 H and 13 C NMR (CDCl 3 ) data agreed with those of literature data [11, 12], so compound 1 was identified as 30-hydroxy-3-friedelanone. Compound 2 was obtained as a white amorphous powder (mp 272–273C), and the molecular ion peak ([M] + , m/z 442) observed in the mass spectra indicated that the molecular formula is C 30 H 50 O 2 . The 1 H and 13 C NMR (CDCl 3 ) data agreed with the literature data [11, 12], so compound 2 was identified as 29-hydroxy-3-friedelanone. Compound 3 was obtained as a white amorphous powder (mp 266–268C) and had the molecular C 30 H 50 O according to the mass spectra ([M + H] + , m/z 427). The 1 H and 13 C NMR (CDCl 3 ) data were in agreement with the literature [13, 14], so

Lupane-type Triterpenoids from the Leaves of Heteropanax fragrans

Lupane-type triterpenoids, a very large and interesting family of compounds, are widely distributed in food, herbs, and other plants throughout the world. This class of compounds possess a wide range of biological properties including cytotoxic, antitumor, antiviral, anti-inflammatory effects and the ability to induce apoptosis in the most prevalent cancer cell lines, such as HepG2, Hep3B and MCF7 cells. The betulinic acid, one of the known lupanetype triterpenoids, showed the selective cytotoxic activity toward melanoma cell lines and is being examined in Phase I trials as a cancer chemoprevention agent. For their various pharmacological and medicinal properties, these pentacyclic triterpenes of the lupane-type have recently been investigated by the scientific community. Heteropanax fragrans (Roxb.) Seem (Araliaceae), an ornamental plant and important source of herb, is distributed in the southern China. The rhizomes of H. fragrans have been used for acesodyne, hemostasia and detumescence by local people. Previous studies reported that four lupane-type triterpenoids and glycosides were isolated from the rhizomes of H. fragrans. However, chemical constituents of the leaves of H. fragrans have not been studied. In this paper, we report the isolation and structure elucidation of one new lupanetype triterpenoid, 3β-hydroxylup-20(29)-ene-23,27,28-trioic acid (1), as well as four known lupane-type derivatives, melaleucic acid (2), 3-oxolup-20(29)-ene-27,28-dioic acid (3), cylicodiscic acid (4), and 3β,23-dihydroxylup-20(29)ene-27,28-dioic acid (5) from the leaves of the title plant (Fig. 1). All the compounds were tested for the cytotoxic activity against KB cell. Compound 1 was obtained as a white amorphous powder. Its molecular formula was assigned as C30H44O7 based on HRESIMS ([M-H]−, m/z 515.2996), C NMR and DEPT data, indicating nine degrees of unsaturation. The IR spectrum of 1 indicated the existence of hydroxyl (3399 cm) and carboxyl groups (1696 cm). Analysis of the H NMR data (Table 1) and the correlations observed in the HSQC experiment showed that the molecule contained four methyl singlets [δH 0.97 (3H, s), 1.18 (3H, s), 1.62 (3H, s), 1.96 (3H,

Isolation and characterization of ardicrenin from Ardisia crenata Sims

A new, effective and economical method to extract ardicrenin from <I>Ardisia crenata</I> Sims collected in the Wolong natural reserve, Sichuan, China, is established. <I>Ardisia crenata</I> Sims powder is counter-current extracted with 80% methanol reflux, decompressively enriched and centrifuged to defat. Supernatant is applied to macroporous resin column (AB-8) with 80% methanol, ardicrenin is isolated by silica gel chromatography with dichlormethane-acetoacetate-methanol (4:1.5:1) washing, and recrystallized in methanol. The final product which proved to be ardicrenin by analytic procedure including Furier transform infrared (FTIR) and ultraviolet spectrum (UV), mass spectroscopy (MS), nuclear magnetic resonance (NMR) and high performance liquid chromatography (HPLC) is white amorphous powder with yield of 1.59 ± 0.02%.

Acaricidal activity of four fractions and octadecanoic acid-tetrahydrofuran-3,4-diyl ester isolated from chloroform extracts of neem (Azadirachta indica) oil against Sarcoptes scabiei var. cuniculi larvae in vitro

Four fractions obtained from chloroform extracts of neem (Azadirachta indica) oil by column chromatography were investigated for acaricidal activity against Sarcoptes scabiei var. cuniculi larvae in vitro. Octadecanoic acid-tetrahydrofuran-3,4-diyl ester was isolated from an active fraction of the chloroform extract and its toxicity against S. scabiei larvae was tested in vitro. A complementary log–log model was used to analyse the toxicity data. Activity was found in the third fraction, with 100% corrected mortality after 4.5h of exposure at a concentration of 200mgml−1. This fraction was repeatedly re-crystallised in acetone to yield a white amorphous powder, identified as octadecanoic acid-tetrahydrofuran-3,4-diyl ester, with a median lethal concentration (LC50) of 0.1mgml−1 at 24h post-treatment. The median lethal time (LT50) for this compound was 15.3h at a concentration of 7.5mgml−1.

Metabolites Produced by Actinomycetes — Antiviral Antibiotics and Enzyme Inhibitors

AbstractFor Abstract see ChemInform Abstract in Full Text.

Metabolites produced by actinomycetes--antiviral antibiotics and enzyme inhibitors

In 1995, we discovered new antiherpetic antibiotics, called fattiviracins. The producing organism was classified as a strain belonging to Streptomyces microflavus. The strain produced at least 13 fattiviracin derivatives (FV-1 to FV-13). Fattiviracins were obtained as a white amorphous powder, and their molecular weights are in the range of 1400 to 1500. They are readily soluble in water, methanol, pyridine, and DMSO, but insoluble in other organic solvents. Fattiviracins have macrocyclic diesters formed by the binding of two trihydroxy fatty acids and two D-glucose residues in the molecule, and they can be divided into five families according to the length of the fatty acid moiety. Fattiviracins have potent activity against enveloped DNA viruses such as the herpes family, HSV-1, and VZV and enveloped RNA viruses such as influenza A and B viruses, and three strains of HIV-1, with EC(50) values on the order of a few micrograms per milliliter. The biosynthetic pathway of fattiviracins is also becoming clearer. Using bacitracin-resistant strains, enhanced and astringent production of fattiviracin was achieved. Fattiviracin FV-13, which has the longest fatty acid chains in the molecule, was dramatically enhanced by a C(55)-isoprenyl phosphate metabolism. In addition, we have screened various inhibitors of enzymes such as alkaline protease, glucosyltransferase, glucuronidase, phospholipase, deoxyribonuclease, DNA methyltransferase, and DNA topoisomerase. All the inhibitors we discovered are briefly summarized in this paper.

Fattiviracins, Antiviral Antibiotics Produced by an Actinomycete

In 1995, we discovered new antiherpetic antibiotics, called fattiviracins. The producing organism was classified as a strain belonging to Streptomyces microflavus. The strain produced at least 13 fattiviracin derivatives (FV-1 to FV-13). Fattiviracins were obtained as a white amorphous powder, and their molecular weights are in the range of 1400 to 1500. They are readily soluble in water, methanol, pyridine and DMSO, but insoluble in other organic solvents. Fattiviracins have macrocyclic diesters formed by the binding of two trihydroxy fatty acids and two D-glucose residues in the molecule, and they can be divided into 5 families, according to the length of fatty acid moiety. Fattiviracins have a potent activity against enveloped DNA viruses such as the herpes family, HSV-1 and VZV and enveloped RNA viruses such as influenza A and B viruses, and three strains of HIV-1, in the order of a few μg/ml of EC50. The biosynthetic pathway of fattiviracins is also becoming clearer. Using bacitracin-resistant strains, enhanced and astringent production of fattiviracin was recognized: fattiviracin FV-13, having the longest fatty acid chains in the molecule was dramatically enhanced by a C55-isoprenyl phosphate metabolism.