Phenolic content and antibacterial activity of extracts of Hamelia patens obtained by different extraction methods

Abstract Hamelia patens, is a plant traditionally used to treat a variety of conditions among the Huastec people of Mexico. The objective of this study is to characterize the phenolic content and critically examine the antimicrobial activity of leaf extracts H. patens, obtained by maceration, Soxhlet and percolation, using ethanol as 70% solvent. Phenolic compounds are characterized by liquid chromatography, coupled to a High Resolution Mass Spectrometry, and the antimicrobial activity was studied from the inhibitory effect of each extract for Escherichia coli, Staphylococcus aureus, Salmonella typhi and S. paratyphi, and by the Minimum Bactericidal Concentration, the percentage of activity and the Index of Bacterial Susceptibility of each extract. The phenolic compound identified in different concentrations in the three extracts was epicatechin. The extracts obtained by the three methods had antimicrobial activity, however, there was no significant difference (p < 0.05) between the Minimum Bactericidal Concentration of the extracts obtained by maceration, percolation and Soxhlet. The results of this study contribute to the body of knowledge on the use of extracts in controlling microorganisms with natural antimicrobials.

Statistical tools application on dextranase production from Pochonia chlamydosporia (VC4) and its application on dextran removal from sugarcane juice

ABSTRACT The aim of this study was to optimize the dextranase production by fungus Pochonia chlamydosporia (VC4) and evaluate its activity in dextran reduction in sugarcane juice. The effects, over the P. chlamydosporia dextranase production, of different components from the culture medium were analyzed by Plackett-Burman design and central composite design. The response surface was utilized to determine the levels that, among the variables that influence dextranase production, provide higher production of these enzymes. The enzymatic effect on the removal of dextran present in sugarcane juice was also evaluated. It was observed that only NaNO3 and pH showed significant effect (p<0.05) over dextranase production and was determined that the levels which provided higher enzyme production were, respectively, 5 g/L and 5.5. The dextranases produced by fungus P. chlamydosporia reduced by 75% the dextran content of the sugarcane juice once treated for 12 hours, when compared to the control treatment.

Actiflagelin, a new sperm activator isolated from Walterinnesia aegyptia venom using phenotypic screening

Sperm contains a wealth of cell surface receptors and ion channels that are required for most of its basic functions such as motility and acrosome reaction. Conversely, animal venoms are enriched in bioactive compounds that primarily target those ion channels and cell surface receptors. We hypothesized, therefore, that animal venoms should be rich enough in sperm-modulating compounds for a drug discovery program. Our objective was to demonstrate this fact by using a sperm-based phenotypic screening to identify positive modulators from the venom of Walterinnesia aegyptia. Methods Herein, as proof of concept that venoms contain interesting compounds for sperm physiology, we fractionated Walterinnesia aegyptia snake venom by RP-HPLC and screened for bioactive fractions capable of accelerating mouse sperm motility (primary screening). Next, we purified each compound from the positive fraction by cation exchange and identified the bioactive peptide by secondary screening. The peptide sequence was established by Edman sequencing of the reduced/alkylated compound combined to LC-ESI-QTOF MS/MS analyses of reduced/alkylated fragment peptides following trypsin or V8 protease digestion. Results Using this two-step purification protocol combined to cell phenotypic screening, we identified a new toxin of 7329.38 Da (actiflagelin) that activates sperm motility in vitro from OF1 male mice. Actiflagelin is 63 amino acids in length and contains five disulfide bridges along the proposed pattern of disulfide connectivity C1-C5, C2-C3, C4- C6, C7-C8 and C9-C10. Modeling of its structure suggests that it belongs to the family of three finger toxins with a noticeable homology with bucandin, a peptide from Bungarus candidus venom. Conclusions This report demonstrates the feasibility of identifying profertility compounds that may be of therapeutic potential for infertility cases where motility is an issue.(AU)

An UPLC-MS/MS method for simultaneous determination of multiple constituents in Guizhi Fuling capsule with ultrafast positive/negative ionization switching / 中国天然药物

Guizhi Fuling capsule (GFC), a traditional Chinese medicine (TCM) with effects of promoting blood circulation and dissipating blood stasis, has been widely used in the clinic. Because of the complex matrix and various chemical structure types, quality control of GFC remains great challenge. In the present study, an ultra performance liquid chromatography hybrid triple-quadrupole mass spectrometry (UPLC-QQQ MS) method with ultrafast positive/negative ionization switching was developed for simultaneous determination of 18 bioactive components in GFC, including methyl gallate, ethyl gallate, oxypaeoniflorin, benzoic acid, albiflorin, paeonolide, paeoniflorin, 1, 2, 3, 4, 6-pentagalloylglucose, mudanpioside C, benzoyloxypaeoniflorin, benzoylpaeoniflorin, pachymic acid, amygdalin, cinnamaldehyde, paeonol, cinnamic acid, 4-hydroxybenzoic acid, and gallic acid. Separation was performed on an Agilent Zorbax Extend-C18 column (2.1 mm × 50 mm, 1.8 μm), using a gradient elution with acetonitrile and water containing 0.1% formic acid. Cholic acid was selected as the internal standard. This newly developed method was fully validated for linearity, precision, accuracy, and stability, and then applied to quality assessment of GFC. Finally, the batch-to-batch reproducibility of GFC samples was evaluated by the cosine ration and Euclidean distance method, which showed high quality consistency. The results demonstrated that the developed method pro vided a reasonable and powerful manner for quality control of GFC.

An UPLC-MS/MS method for simultaneous determination of multiple constituents in Guizhi Fuling capsule with ultrafast positive/negative ionization switching / 中国天然药物

Guizhi Fuling capsule (GFC), a traditional Chinese medicine (TCM) with effects of promoting blood circulation and dissipating blood stasis, has been widely used in the clinic. Because of the complex matrix and various chemical structure types, quality control of GFC remains great challenge. In the present study, an ultra performance liquid chromatography hybrid triple-quadrupole mass spectrometry (UPLC-QQQ MS) method with ultrafast positive/negative ionization switching was developed for simultaneous determination of 18 bioactive components in GFC, including methyl gallate, ethyl gallate, oxypaeoniflorin, benzoic acid, albiflorin, paeonolide, paeoniflorin, 1, 2, 3, 4, 6-pentagalloylglucose, mudanpioside C, benzoyloxypaeoniflorin, benzoylpaeoniflorin, pachymic acid, amygdalin, cinnamaldehyde, paeonol, cinnamic acid, 4-hydroxybenzoic acid, and gallic acid. Separation was performed on an Agilent Zorbax Extend-C18 column (2.1 mm × 50 mm, 1.8 μm), using a gradient elution with acetonitrile and water containing 0.1% formic acid. Cholic acid was selected as the internal standard. This newly developed method was fully validated for linearity, precision, accuracy, and stability, and then applied to quality assessment of GFC. Finally, the batch-to-batch reproducibility of GFC samples was evaluated by the cosine ration and Euclidean distance method, which showed high quality consistency. The results demonstrated that the developed method pro vided a reasonable and powerful manner for quality control of GFC.

Genotoxicidad de los hidrocarburos aromáticos policíclicos extraídos mediante el sistema diclorometano-etanol-tolueno en muestras del aire de Cúcuta, Norte de Santander, Colombia / Genotoxicity of polycyclic aromatic hydrocarbons extracted by dichloromethane-ethanol-toluene system air samples from Cúcuta, Norte de Santander, Colombia

Los contaminantes del aire han sido y siguen siendo, los principales factores que contribuyen a las enfermedades crónicas como el asma y enfermedades cardiovasculares. La contaminación del aire por material particulado (PM) es un problema mundial y en los últimos años, el PM se ha convertido en un tema importante de investigación ya que tiene un impacto negativo significativo en la salud humana; el PM es generado por las actividades industriales y tubos de escape de vehículos de motor. Sin embargo, diversos componentes nocivos del PM, como los hidrocarburos aromáticos policiclicos (HAP) en general, son sos­pechosos de ser carcinogénicos. Este trabajo tiene como objetivo identificar los HAP presentes en el PM2.5 del aire de Cúcuta, extraídos por primera vez, mediante el sistema diclorometano-etanol-tolueno e investigar la importancia del fraccionamiento de la materia organica del PM2.5 para detectar los HAP presentes en las fracciones del PM2.5. La identificación de los HAP considerados como contaminantes prioritarios y reconocidos por su afectación a la salud de la población se realizó, mediante cromatografía de gases con detector FID. Los efectos genotoxicos de la materia orgánica del PM2.5 extraída con una mezcla de DCM-etanol-tolueno fueron evaluados mediante el ensayo Cometa.

Air pollutants have been and still are the main factors that contribute to chronic diseases such as asthma and cardio­vascular disease. Air pollution by particulate matter (PM) is a global problem and in recent years, the PM has become an important research topic since it has a significant negative impact on human health; the PM is generated by industrial activities and exhaust pipes of motor vehicles. However, various harmful components of PM such as polycyclic aromatic hydrocarbons (PAHs) in gen­eral, are suspected of being carcinogenic. This work aims to identify the PAHs present in the PM 2.5 air Cúcuta, first extracted by the dichloromethane-ethanol-toluene system and investigate the importance of organic matter fractionation of PM 2.5 to detect PAHs present in the fractions of PM 2.5. The identification of PAHs considered as priority pollutants and recognized for their effects on health of the population was performed by gas chromatography with FID detector. The genotoxic effects of PM2.5 organic mat­ter, extracted with a mixture of DCM-ethanol-toluene, was evaluated by the Comet assay.

Fraccionamiento y caracterización electroforética de las proteínas de la semilla de kañihua (Chenopodium pallidicaule Aellen) / Fractionation and electrophoretic characterization of (Chenopodium pallidicaule Aellen) kanihua seed proteins

Kanihua (Chenopodium pallidicaule Aellen) is a Chenopodiacea of the andean region, that contains between 15 and 19% protein, with essential amino acids. The objective of the study was to fractionate and electrophoretically characterize the proteins of kanihua seed varieties Cupi-Sayhua and Ramis. In the whole meal, the proximal analysis and fractionation were performed, and the flour was fractionated by five techniques according to Osborne solubility to obtain albumins, globulins, prolamins andglutelins. The methodology, solvents and extraction time were optimized; and the electrophoretic profiles of the fractions were identified. The highest protein content (p≤ 0.05) was of kanihua flour and its protein fractions, compared to kiwicha and wheat. The highest percent yield (p≤ 0.05) during 1 h of sequential extraction of the protein fractions, was obtained with the Rodriguez y et.al., technique for albumins and glutelins, and with the technique described by Barba de la Rosa y et.al., for globulins and prolamins. The following results were foundin Ramis and Cupi-Sayhua kanihua: albumins: 15.4±0.3 and 15.8±0.3%, globulins 7S: 24.1±0.5 and 26.3±1.0%, globulins 11S: 25.7±1.0 and 26.7±1.0%, prolamins: 9.6±0.1 and 9.9±0.5% and glutelins: 22.9±0.1 and 21.5±1.4%, respectively The electrophoretic profile showedpatterns similar in number of bands and differences in concentration in both varieties.

La kañihua (Chenopodium pallidicaule Aellen) es una Chenopodiacea de la región andina, que contiene entre 15 y 19% de proteínas, con aminoácidos esenciales. El objetivo del presente estudio fue fraccionar y caracterizar electroforéticamente las proteínas de la semilla de kañihua variedades Ramis y Cupi-Sayhua. En harina integral se realizó el análisis proximal y fraccionamiento, luego, la harina se fraccionó mediante cinco técnicas según la solubilidad de Osborne para obtener albuminas, globulinas, prolaminas y glutelinas. Se optimizó la metodología, solventes y tiempo de extracción; e identificaron los perfiles electroforéticos de las fracciones. El mayor contenido proteínico (p≤ 0,05) fue de la harina de kañihua y sus fracciones proteicas, en comparación a kiwicha y trigo. El mayor rendimiento porcentual (p≤ 0,05) durante 1 h de extracción secuencial de las fracciones proteicas, se obtuvo con la técnica de Rodríguez y et.al., para albuminas y glutelinas, y con la técnica de Barba de la Rosa y et.al., para globulinas y prolaminas. Se encontró en kañihua Ramis y Cupi-Sayhua, albúminas: 15,4±0,3 y 15,8+0,3%; globulinas: 7S 24,1±0,5 y 26,3+1,0%; globulinas 11S: 25,7+1,0 y 26,7+1,0%; prolaminas: 9,6+0,1 y 9,9+0,5% y glutelinas: 22,9+0,1 y 21,5+1,4%, respectivamente. El perfil electroforético mostró patrones similares en número de bandas y diferentes en concentración en ambas variedades.

Isoflavone formononetin from red propolis acts as a fungicide against Candida sp

Abstract A bioassay-guided fractionation of two samples of Brazilian red propolis (from Igarassu, PE, Brazil, hereinafter propolis 1 and 2) was conducted in order to determine the components responsible for its antimicrobial activity, especially against Candida spp. Samples of both the crude powdered resin and the crude ethanolic extract of propolis from both locations inhibited the growth of all 12 tested Candida strains, with a minimum inhibitory concentration of 256 µg/mL. The hexane, acetate and methanol fractions of propolis 1 also inhibited all strains with minimum inhibitory concentration values ranging from 128 to 512 µg/mL for the six bacteria tested and from 32 to 1024 µg/mL for the yeasts. Similarly, hexane and acetate fractions of propolis sample 2 inhibited all microorganisms tested, with minimum inhibitory concentration values of 512 µg/mL for bacteria and 32 µg/mL for yeasts. The extracts were analyzed by HPLC and their phenolic profile allowed us to identify and quantitate one phenolic acid and seven flavonoids in the crude ethanolic extract. Formononetin and pinocembrin were the major constituents amongst the identified compounds. Formononetin was detected in all extracts and fractions tested, except for the methanolic fraction of sample 2. The isolated isoflavone formononetin inhibited the growth of all the microorganisms tested, with a minimum inhibitory concentration of 200 µg/mL for the six bacteria strains tested and 25 µg/mL for the six yeasts. Formononetin also exhibited fungicidal activity against five of the six yeasts tested. Taken together our results demonstrate that the isoflavone formononetin is implicated in the reported antimicrobial activity of red propolis.

A new biocompatible microemulsion increases extraction yield and bioavailability of Andrographis paniculata / 中国天然药物

The purpose of this study was to design and prepare a biocompatible microemulsion of Andrographis paniculata (BMAP) containing both fat-soluble and water-soluble constituents. We determined the contents of active constituents of BMAP and evaluated its bioavailability. The biocompatible microemulsion (BM), containing lecithin and bile salts, was optimized in the present study, showing a good physical stability. The mean droplet size was 19.12 nm, and the average polydispersity index (PDI) was 0.153. The contents of andrographolide and dehydroandrographolide in BMAP, as determined by high performance liquid chromatography (HPLC), were higher than that in ethanol extraction. The pharmacokinetic results of BMAP showed that the AUC0-7 and AUC0→∞ values of BMAP were 2.267 and 27.156 μg·mL(-1)·h(-1), respectively, and were about 1.41-fold and 6.30-fold greater than that of ethanol extraction, respectively. These results demonstrated that the bioavailability of and rographolide extracted by BMAP was significantly higher than that extracted by ethanol. In conclusion, the BMAP preparation displayed ann improved dose form for future clinical applications.

Antioxidant activities of crude phlorotannins from Sargassum hemiphyllum / 华中科技大学学报（医学）（英德文版）

Brown algae are well known as a source of biologically active compounds, especially those having antioxidant activities, such as phlorotannins. In this study we examined the antioxidant activities of crude phlorotannins extracts (CPEs) obtained from Sargassum hemiphyllum (SH) and fractionated according to the molecular weights. When CPEs were administrated at a dose of 30 mg/kg to Kunming mice pre-treated with carbon tetrachloride (CCl4), the levels of oxidative stress indicators in the liver, kidney and brain were significantly reduced in vivo. All the components of various molecular weight fractions of CPEs exhibited greater scavenging capacities in clearing hydroxyl free radical and superoxide anion than the positive controls gallic acid, vitamin C and vitamin E. Particularly, the components greater than 30 kD obtained from ethyl acetate phase showed the highest antioxidant capacities. These results indicated that SH is a potential source for extracting phlorotannins, the algal antioxidant compounds.

Synergistic, additive and antagonistic effects of fruit mixtures on total antioxidant capacities and bioactive compounds in tropical fruit juices / Efectos sinérgicos, aditivos y antagónicos de mezclas de pulpa frutas en la capacidad antioxidante total y compuestos bioactivos en los zumos de frutas tropicales

The objective of this work was investigate the synergistic, additive and antagonistic effects of fruit mixtures on total antioxidant capacities and bioactive compounds in tropical fruit juices, and optimize its formulation by the response surface methodology based on the responses: total polyphenols (TP), total antioxidant capacity (TAC), ascorbic acid content and sensorial acceptance. Camu-camu, acerola and acai were the major factors that influenced the antioxidant potential of the juice; and the yellow mombin showed a positive effect on the acceptance of the tropical juice. It was observed an antagonistic effect between acerola and camu-camu for the TAC response. The optimum formulation obtained was 20% acerola, 10% camu-camu, 10% yellow mombin, 10% cashew apple and 10% acai, which was responsible for a response of 155.46 mg.100 g-1 of ascorbic acid, 103.01 mg of GAE.100 g-1 of TP, 10.27 μM Trolox g-1 of TAC and approximately 6.1 of acceptance.

El objetivo de este trabajo fue investigar los efectos sinérgicos, aditivos y antagónicos de mezclas de diferentes frutas tropicales en la capacidad antioxidante total (TAC) y compuestos bioactivos presentes en los jugos mixtos, y optimizar su formulación por la metodología de superficie de respuesta basado en las evaluaciones de: polifenoles totales (TP), capacidad antioxidante total (TAC), contenido de ácido ascórbico y la aceptación sensorial. Camu-camu, acerola y acai fueron las frutas que más influyeron en el potencial antioxidante del jugo mixto; y el jobo mostró un efecto positivo en la aceptación del jugo mixto tropical. Se observó un efecto antagónico entre acerola y camu-camu para la TAC. La formulación óptima obtenida contenía 20% acerola, 10% de camu-camu, 10% el jobo, 10% de manzana de marañón y 10% de acai, la cual ha proporcionado contenidos medio de 155,46 mg.100 g-1 de ácido ascórbico, 103,01 mg de GAE.100 g-1 de TP, 10,27 mM Trolox g-1 de TAC y aproximadamente 6.1 de aceptación sensorial.

Identification and characterization of a potent anticancer fraction from the leaf extracts of Moringa oleifera L.https://docs.google.com/forms/d/1RkQLeYfiMvE7SKcdOXv9Acm5eXRjmvleic5FYjjX1TE/viewform?fbzx=6173639259593088622

Anticancer potential of Moringa oleifera L. extracts have been well established. However, there are no reports on the isolated molecules/fractions from these extracts which are responsible for the anticancer/cytotoxic activity. Thus, in the present study, we explored the same. The n-hexane, chloroform, ethyl acetate, methanol extracts of the M. oleifera leaves and 15 fractions (F1 to F15) of ethyl acetate extract were evaluated for their in vitro and in vivo anticancer activity using Hep-2 cell lines and Dalton’s lymphoma ascites model in mice, respectively. Among the tested samples, the F1 fraction showed potential cytotoxic effect in Hep-2 cell lines with a CTC50 value of 12.5 ± 0.5 µg/ml. In vivo studies with the doses 5 and 10 mg/kg, p.o. demonstrated significant reduction in body weight and increased the mean survival time compared to the control group. These results were also comparable to the standard, 5-Fluorouracil, treated animals. We have also successfully isolated and characterized the anticancer fraction, F1 from the leaves of M. oleifera L.

Purification technology of manninotirose in Rehmanniae Radix Praeparata by D-101 microporous adsorption resin / 中国中药杂志

This paper was aim to optimize the purification technology of Rehmanniae Radix Praeparata extract with macroporous adsorption resin. With the content of manninotriose as index, the absorptive flow and time were investigated, as well as kinds, amount, flow of eluent. D-101 type macroporous adsorption resin was the best choice for the purification of manninotriose. The optimized parameters were as follows: the content of manninotriose at 161.16-53.72 mg x g(-1), absorption time 240 min, eluting solvent of purified water, volume flow at 1.5 BV x h(-1), and eluant volume at 6 BV. D-101 type macroporous adsorption resin could significantly increase the purity of Rehmanniae Radix Praeparata extract with the advantage of high absorption, remove most part of impurity, and the effect of semi-works production was better.

Chemical constituents from EtOAc fraction of Sophora dunnii / 中国中药杂志

Sixteen compounds have been isolated from the EtOAc fraction of 95% ethanolic extract of Sophora dunnii through silica gel, Sephadex LH-20 and semi-prerarative HPLC column chromatographies. Their structures were identified on the basis of NMR and MS spectra data as phaseollidin (1), L-maackiain (2), 2-(2',4'-dihidroxyphenyl)-5,6-methylenedioxy benzofuran (3), 8-demethyl-farrerol (4), liquiritigenin (5), genistein (6), 6-methylgenistein (7), 5-O-methyl genistein (8), 7,2',4'-trihydroxys-5-methoxy-isoflavanone (9), 7, 3', 4'-trihydroxy-isoflavanone (10), erythribyssin D (11), calycosin (12), trans-resveratrol (13), cis-resveratrol (14), stigmasterol (15), β-sitosterol (16). Among these, compounds 1-14 and 16 were isolated from this plant for the first time.

Optimization of extraction technology for salidroside, tyrosol, crenulatin and gallic acid in Rhodiolae Crenulatae Radix et Rhizoma with orthogonal test / 中国中药杂志

The extracting technology of salidroside, tyrosol, crenulatin and gallic acid from Rhodiolae Crenulatae Radix et Rhizoma was optimized. With extraction rate of salidroside, tyrosol, crenulatin and gallic acid as indexes, orthogonal test was used to evaluate effect of 4 factors on extracting technology, including concentration of solvent, the dosage of solvent, duration of extraction, and frequency of extraction. The results showed that, the best extracting technology was to extract in 70% alcohol with 8 times the weight of herbal medicine for 2 times, with 3 hours once. High extraction rate of salidroside, tyrosol, crenulatin and gallic acid were obtained with the present technology. The extracting technology was stable and feasible with high extraction rate of four compounds from Rhodiolae Crenulatae Radix et Rhizoma, it was suitable for industrial production.

Analysis of triterpenoids in Ganoderma lucidum by microwave-assisted continuous extraction / 中国中药杂志

For further improving the extraction efficiency of microwave extraction, a microwave-assisted contijuous extraction (MACE) device has been designed and utilized. By contrasting with the traditional methods, the characteristics and extraction efficiency of MACE has also been studied. The method was validated by the analysis of the triterpenoids in Ganoderma lucidum. The extraction conditions of MACE were: using 95% ethanol as solvent, microwave power 200 W and radiation time 14.5 min (5 cycles). The extraction results were subsequently compared with traditional heat reflux extraction ( HRE) , soxhlet extraction (SE), ultrasonic extraction ( UE) as well as the conventional microwave extraction (ME). For triterpenoids, the two methods based on the microwaves (ME and MACE) were in general capable of finishing the extraction in 10, 14.5 min, respectively, while other methods should consume 60 min and even more than 100 min. Additionally, ME can produce comparable extraction results as the classical HRE and higher extraction yield than both SE and UE, however, notably lower extraction yield than MASE. More importantly, the purity of the crud extract by MACE is far better than the other methods. MACE can effectively combine the advantages of microwave extraction and soxhlet extraction, thus enabling a more complete extraction of the analytes of TCMs in comparison with ME. And therefore makes the analytic result more accurate. It provides a novel, high efficient, rapid and reliable pretreatment technique for the analysis of TCMs, and it could potentially be extended to ingredient preparation or extracting techniques of TCMs.

Simultaneously preparation of grams of high purity tyrosol, crenulatin and salidroside from Rhodiola crenulata / 中国中药杂志

Tyrosol, crenulatin and salidroside are the main active constituents of Rhodiola crenulata, with extensive pharmacological activities. In the study, grams of high purity tyrosol, crenulatin and salidroside were simultaneously separated from R. crenulata by the first time. Firstly, R. crenulata was extracted by 70% alcohol. Then, with the yields of three compounds as the index, the macroporous resin was optimized. At last, grams of high purity tyrosol, crenulatin and salidroside were isolated by D-101 macroporousresin, purified by column chromatography. Detected by HPLC, the purity of three compounds were higher than 98%. This method has the advantages of simple process and operation, less dosage of organic solvent, highly yield and reproducibility, suitable for the simultaneously preparation of tyrosol, crenulatin and salidroside.

Analysis of four flavonoids in Lysimachia clethroides using ionic liquid-assisted extraction / 中国中药杂志

In order to established a method for simultaneous determination of isoquercitrin, astragaline, quercetin and kaempferol in Lysimachia clethroides, the ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate ([HMIM]PF6) methanol was used as the ultrasound-assisted extraction solvent combing with RP-HPLC. A Purospher star RP-C1 column was used with the mobile phase of aceto- nitrile, methanol and 0. 4% phosphate acid by gradient elution at the detection wavelength of 360 nm. The flow rate was 0.7 mL x min(-1), and the column temperature was the room temperature. Under the optimized conditions, the linear ranges were 2.54 x 10(-2)-2. 54, 2.50 x 10(-2)- 2.50, 1.54 x 10(-3)-0.154, 1.49 x 10(-3)-0.149 microg for isoquercitrin, astragaline, quercetin and kaempferol, respectively. The average recoveries of the four constituents were 101.1%, 98.90%, 101.0%, 101.6%, respectively. The method was green, simple, rapid and accurate, and provided a valid method for analysis of isoquercitrin, astragaline, quercetin and kaempferol in L. clethroides.

Investigation of elemental concentration and extraction rate of polysaccharides from Liuwei Dihuang prescription" by ICP-MS / 中国中药杂志

A method was established for the simultaneous analysis of 25 trace elements and heavy metals in polysccharides from Liuwei Dihuang prescription, including Li, Be, B, Ti, Mg, Al, V, Cr, Mn, Co, Fe, Ni, Cu, Zn, Ga, As, Sr, Cd, Sn, Sb, Ba, Hg, Tl, Pb, Bi. The different rate of elemental extraction in Al, Fe, Mg, B, Ti, Mn, Zn, Sr, Ba was made in water and different concentration of alcohol. The samples, digested via microwave, calibrated by internal standard elements such as Ge and In, with bush branches and leaves as the controlled reference standard, were inlet into ICP-MS to analyze the contents of the 24 trace elements and heavy metals. The detection limits of the 24 elements were in the range of 0.007-2.225 µg · L(-1), while the RSD was below ≤ 4. 0%, with their recovery ranging from 84. 1% to 116%. Big different of the elemental extraction rates could be found by using different ethanol solutions. The method is simple, rapid and accurate, and can be used for the quality control of trace elements and heavy metals in Liuwei Dihuang polysccharides. With the aid of the obtained result, we may increase the extraction of necessary element while making an attempt at multi-element speciation in polysccharides from Liuwei Dihuang.

Optimization of extraction technology of compatibility of couplet medicines Epimedium brevicornu and Ligusticum chuanxiong by orthogonal design / 中国中药杂志

The study was using the orthogonal test and making the extraction rates of icariin, ferulic acid, epimedin A, epimedin B, epimedin C, baohuoside I and ligustilide determinated by HPLC multiwavelength switch, gradient elution and multi-index comprehensive weighted scoring method (weight coefficient was 0.47: 0.16: 0.07: 0.07: 0.08: 0.06: 0.09) as evaluation index, combine with SPSS 16.0 software to optimizing the best extraction. It was Yinpian soak 1 h, 12 times more than the volumn of 50% ethanol solution, by heating reflux extraction for 60 min. The compliance test indicates that the optimized compatibility extraction technology is stable and practical, and it has provided an experimental basis for compound preparation technology research of Epimedium brevicornu and Ligusticum chuanxiong.