Purification process of epimedin and icariin from Epimedii Folium by macroporous resin / 中草药

Objective: To investigate the best technological conditions for the purification of epimedin and icariin from Epimedii Folium by macroporous resin, and preliminarily characterize the purification fraction of the best technological conditions. Methods: Five kinds of macroporous resins were screened by static adsorption experiment with the content of epimedin A1, epimedin A, epimedin B, epimedin C and icariin as indexes. The best purification conditions were optimized by the concentration of upper column solution, the maximum sample volume, the upper column flow rate, the volume of water washing, the concentration of removing impurity ethanol and elution ethanol, the volume of removing impurity ethanol and elution ethanol, the column diameter-height ratio through dynamic adsorption experiment. Finally, UPLC-Q-TOF/MS, HPLC and ultraviolet spectrophotometry were used to characterize the purification fraction of the best technological conditions. Results: The best macroporous resin was AB-8, column diameter-height ratio was 1:7, 6 BV of upper column solution (crude drug 0.5 g/mL) was used for dynamic adsorption at a flow rate of 6 BV/h, 5 BV of water and 5 BV of 20% ethanol were used for impurity removal, and 6 BV of 50% ethanol was used for elution. The flow rate of impurity removal and elution was 6 BV/h. After purification, the total flavonoids content was 63.29%, the total content of epimedin A1, A, B, C and icariin was 40.48%, the content of epimedin A1, epimedin A, epimedin B, epimedin C and icariin was 1.63%, 2.52%, 16.36%, 5.51% and 14.46%, respectively. Conclusion: The purification process of epimedin and icariin from Epimedii Folium by AB-8 macroporous resin is stable, reasonable and feasible. The chemical characterization indicated that the purification fraction was mainly flavonoids, mainly consisting of epimedin and icariin. The optimized purification process can be used for the purification and enrichment of such ingredients.

Isolation of flavonol rhamnosides from Pometia pinnata leaves and investigation of α-glucosidase inhibitory activity of flavonol derivatives

Pometia pinnata belonging to the Sapindaceae family has been traditionally used as the therapeutic agent for burns andwounds in Indonesia. Based on the result of the experiment conducted, the ethyl acetate fraction of P. pinnata leavesshowed strong α-glucosidase inhibitory activity. After two flavonol rhamnoside compounds were isolated from ethylacetate fraction of P. pinnata leaves methanol extract using chromatography method, their structures were identifiedas kaempferol-3-O-rhamnoside (1) and quercetin-3-O-rhamnoside (2). The ultra-performance liquid chromatographyelectrospray ionization time-of-flight mass spectrometry (UPLC-ESI-TOFMS) chromatogram showed compounds1 and 2 were the major compounds of the ethyl acetate fraction. In this study, the structure–activity relationshipamong the kaempferol, quercetin, and their derivatives bearing sugar moiety were also evaluated. Among tested eightcompounds, kaempferol 7 (percent inhibition = 80.10% ± 0.8) and quercetin 8 (percent inhibition = 82.93% ± 0.4) hadstronger α-glucosidase inhibitory activity than that of other derivatives. Among kaempferol derivatives bearing sugarmoiety, compound 1 showed the strongest activity. Moreover, compound 2 showed strong α-glucosidase inhibitoryactivity among quercetin derivatives. Therefore, it can be confirmed that the hydroxyl group at C-3 position is veryimportant for α-glucosidase inhibitory activity of flavonol compounds.

Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry for identification of Clostridium species isolated from Saudi Arabia

Abstract The aim of this study was to identify different Clostridium spp. isolated from currency notes from the Ha’il region of Saudi Arabia in September 2014 using MALDI–TOF-MS. Clostridium spp. were identified by Bruker MALDI–TOF-MS and compared with VITEK 2. The confirmation of the presence of different Clostridium spp. was performed by determining the sequence of the 16S ribosomal RNA gene. In this study, 144 Clostridium spp. were isolated. Among these specimens, MALDI–TOF-MS could identify 88.8% (128/144) of the isolates to the species level and 92.3% (133/144) to the genus level, whereas, VITEK 2 identified 77.7% of the (112/144) isolates. The correct identification of the 144 isolates was performed by sequence analysis of the 500 bp 16S rRNA gene. The most common Clostridium spp. identified were Clostridium perfringens (67.36%), Clostridium subterminale (14.58%), Clostridium sordellii (9%) and Clostridium sporogenes (9%). The results of this study demonstrate that MALDI–TOF-MS is a rapid, accurate and user friendly technique for the identification of Clostridium spp. Additionally, MALDI–TOF-MS has advantages over VITEK 2 in the identification of fastidious micro-organisms, such as Clostridium spp. Incorporating this technique into routine microbiology would lead to more successful and rapid identification of pathogenic and difficult to identify micro-organisms.

Study on Sini decoction in treatment of hypothyroidism by metabonomic method / 药学实践杂志

Objective To evaluate therapeutic effects of Sini decoction (SND) on hypothyroidism based on serum meta-bonomics method .Methods Twenty-four rats were randomly divided into three groups including sham group ,surgery-induced hypothyroid model group and SND-treated group .The SND-treated group was given SND for 4 weeks .After 4 weeks ,the ser-um samples of each rat were collected and analyzed by UHPLC-Q-TOFMS ,pattern recognition analysis of metabolomics differ-ences among the groups and therapeutic effects of SND were evaluated .Results Total of 9 metabolites were identified through serum metabonomics analysis .It was showed that there was a possible mechanism that hypothyroidism was mainly involved in citrate cycle ,phenylalanine metabolism ,sphingolipid metabolism and phospholipid metabolism and the damage of hypothyroid-ism was reversed by SND .Conclusion SND administration could provide satisfactory effects on hypothyroidism through par-tially regulating the perturbed citrate cycle ,phenylalanine metabolism and phospholipid metabolism .

Intervention effects of Astragalus saponins on spleen deficiency syndrome rats based on metabonomic research / 中草药

Objective: Effect of Astragalus saponins on the changes of endogenous metabolites in plasma of rats with spleen deficiency syndrome of dampness and the intervention function of Astragalus Radix saponins on the syndrome was studied. Methods: High performance liquid phase of flight time mass spectrometry was used to detect the information of endogenous metabolites in rat plasma, principal component analysis and partial least-squares discriminate analysis were used to study the metabolic differences between normal group, model group, and astragalus saponins group to identify potential biomarkers. Results: The metabolism pattern of normal group, model group, and Astragalus saponin group were significantly different, 11 biomarkers were identified and the pathway mainly involved glycerol phospholipids metabolism, sphingolipid metabolism, pentose glucuronic acid conversion, and peanut arachidonic acid metabolism. Conclusion: The results of this study show that it is possible for Astragalus Radix saponins group to interfere with the spleen deficiency syndrome of dampness by regulating energy metabolism, lipid metabolism, and fatty acid metabolism to exert therapeutic effect.

Polymer impurities in cefotiam hydrochloride by online 2D-GFC-RPLC-TOFMS / 中国药学杂志

OBJECTIVE: To validate the conventional gel filtration chromatography (GFC) method for determination of the polymer impurities in β-lactam antibiotics and achieve combination of GFC and RP-HPLC system. METHODS: The effectiveness of the conventional GFC method was identified by 2D-GFC X LC-TOFMS and the polymer impurities found by GFC were identified by RPHPLC. RESULTS: The polymer impurities found by GFC mainly were degradation impurities and open β-lactam ring impurities, which could not effectively characterize the sensitizing polymer impurities in cefotiam hydrochloride. CONCLUSION: An effective method to characterize the polymer impurities in β-lactam antibiotics may be established on the basis of online column switching technique which effectively combines the advantages of GFC and the ability of RP-HPLC to identify the special impurities.

Rapid identification of chemical components in Artemisiae argyi folium by RRLC-TOFMS / 药学实践杂志

Objective To identify the chemical components in Artemisiae argyi folium by rapid resolution liquid chromatog-raphy-time of flight mass spectrometry (RRLC-TOFMS).Methods The separation was performed on an Agilent Eclipse C18 column (2.1 mm ×100 mm,1.8 μm ).The mobile phase consisted of acetonitrile (A) and 0.1% formic acid (B) were in gradient elu-tion.The flowing rate was 0.35 ml/min, the injection volume was 1μl and the temperature of column was 40℃.Time of flight mass spectrometer ( TOFMS) with electro spray ion source ( ESI) was applied to qualitative analysis under the positive ion mode, and mass scan range was m/z 100-1 500 .Results 31 chemical compounds in Artemisiae argyi folium were identified unequivocally .Conclu-sion A rapid and efficient RRLC-TOFMS approach for identifying the chemical constituents of Artemisiae argyi folium had been suc-cessfully established,which paved a way for quality control and further in vivo studies of Artemisiae argyi folium.

Analysis on chemical constitutes in Psoraleae Fructus by combination of HPLC/TOF-MS and HPLC/IT-MSn / 中草药

Objective: To establsh a combinative HPLC/TOF-MS and HPLC/IT-MSn method for quantitative analysis on chemical constituents in Psoraleae Fructus. Methods: Separation was performed on a Phenomenex Luna C18 (5 μm) reverse-phase column. The mobile phase consisted of water containing 0.1% formic acid and acetonitrile was used as gradient elute. The flow rate was 1 mL/min. A combination of time-of-flight mass spectrometer (TOF-MS) and ion trap tandem mass spectrometer (IT-MSn) was applied for qualitative analysis under positive and negative ion mode. Results: Under optimized LC/MS condition, the molecular formulae of 13 components were screened by TOF-MS and all of them were then identified by IT-MSn. Conclusion: The established combinative HPLC/MS method could thus be considered as an effective and credible pattern for the analysis on the multi-components of Chinese materia medica and it also could be used for as the analysis on other complex systems.

Liquid chromatography coupled with time-of-flight and ion trap mass spectrometry for qualitative analysis of herbal medicines / 药物分析学报

With the expansion of herbal medicine (HM) market, the issue on how to apply up-to-date analytical tools on qualitative analysis of HMs to assure their quality, safety and efficacy has been arousing great attention. Due to its inherent characteristics of accurate mass measurements and multiple stages analysis, the integrated strategy of liquid chromatography (LC) coupled with time-of-flight mass spectrometry (TOF-MS) and ion trap mass spectrometry (IT-MS) is well-suited to be performed as qualitative analysis tool in this field. The purpose of this review is to provide an overview on the potential of this integrated strategy, including the review of general features of LC-IT-MS and LC-TOF-MS, the advantages of their combination, the common procedures for structure elucidation, the potential of LC-hybrid-IT-TOF/MS and also the summary and discussion of the applications of the integrated strategy for HM qualitative analysis (2006-2011). The advantages and future developments of LC coupled with IT and TOF-MS are highlighted.

Identification of homoisoflavonoids in Ophiopogon japonicus alcohol extract by an LC-MS based on precise mass and tandem mass spectrometer / 中草药

Objective: To develop an LC/MS approach based on precise mass and tandem mass spectrometer to identify homoisoflavonoids from Ophiopogon japonicus alcohol extract. Methods: Separation was performed on an Alltima C18 reverse-phase column (250 mm x 4.6 mm, 5 μm) and an Alltech pre-column. The mobile phase consisted of water containing 1 mmol/L ammonium acetate-acetonitrile containing 0.01% acetic acid was used as gradient elution. The flow rate was 1.0 mL/min. The detection wavelength was 298 nm. A combination of time-of-flight mass spectrometer (TOF/MS) and ion trap tandem mass spectrometer (Trap/MSn) was applied for qualitative analysis under negative ion mode. Results: From the alcohol extract of O. japonicus, the molecular formulae of 11 components were screened by TOF/MS and 11 chemical components were identified by Trap/MSn. All of the peaks were attributed. Conclusion: This method can be used to identify medicinal ingredients quickly and provide an effective and reliable analytical model, as well as other complex system of traditional Chinese medicine compound.

HPLC-TOFMS in rapid separation and identification of chemical components in Oldenlandia diffusa and its injection preparations / 第二军医大学学报

Objective To rapidly separate and identify the chemical components in traditional Chinese herbal medicine of Oldenlandia diffusa and its injection preparations by high performance liquid chromatography-time of flight mass spectrometry (HPLC-TOFMS). Methods An Agilent Zorbax XDB-Q18 column (250 mm×4. 6 mm, 5 μm) was used for separation and identification of chemical components in Oldenlandia diffusa, with a mobile phase of 0. 3% acetic acid (A) and methanol (B) in gradient elution, 0-30 min, 30%-90%B. The flow rate was set at 1. 0 ml/min and the injection volume was 10 μl. The time- of-flight mass spectrometer was equipped with an EIS ion source. The scanning mass range was between m/z 100-1 000. Results The traditional Chinese medicine of Oldenlandia diffusa and its injection preparation were on-line separated and characterized by HPLC-TOFMS, and 11 chemical compounds were identified in Oldenlandia diffusa, 6 compounds in market injection preparation, and 2 compounds in the injection prepared by ourselves. Conclusion Chromatographic demonstration of chemical compounds in Oldenlandia diffusa in one run provides a foundation for the further studying the metabolism and mechanism of Oldenlandia diffusa and its injection preparations.

Rapid analysis of components in Rhizoma Anemarrhenae by HPLC-DAD-MS and HPLC-DAD-TOFMS / 西安交通大学学报·英文版

A global quality control method based on high performance liquid chromatography (HPLC) coupled with diode array detection (DAD), single quadrupole mass spectrometry (MS) and time-of-flight mass spectrometry (TOFMS) was developed for simultaneous determination of seven major components (mangiferin, neomangiferin, timosaponin E1, timosaponin E, timosaponin BII, timosaponin BIII, and timosaponin A III) and identification of most components in extracts of Rhizoma Anemarrhenae (RA). HPLC analysis was performed on an Agilent SB-C18 column (4.6 mm X 150 mm, 5 μm) by gradient elution using acetonitrile and water-acetic acid(100:0.05, v/v) as the mobile phase. Seven major components in RA were successfully separated. This quantitative method was fully validated in respect of the following performance criteria, linearity, precision, repeatability, stability, accuracy, limits of detection (LOD) and quantification (LOQ). A formula database of known compounds in RA was established, against which, most of the reported components in this herbal extract were identified effectively based on the extract masses acquired by TOFMS. This qualitative and quantitative method was successfully used to analyze the components in 10 batches of RA samples collected from different regions in China. This global quality control method, which consisted of HPLC-DAD-MS assay of seven major components and unambiguous identification of nineteen components, is suitable for routine quantification and comprehensive quality control of RA.

Rapid analysis of components in Rhizoma Anemarrhenae by HPLC-DAD-MS and HPLC-DAD-TOFMS / 药物分析学报

A global quality control method based on high performance liquid chromatography (HPLC) coupled with diode array detection (DAD), single quadrupole mass spectrometry (MS) and time-of-flight mass spectrometry (TOFMS) was developed for simultaneous determination of seven major components (mangiferin, neomangiferin, timosaponin E1, timosaponin E, timosaponin BⅡ, timosaponin BⅢ, and timosaponin AⅢ) and identification of most components in extracts of Rhizoma Anemarrhenae (RA). HPLC analysis was performed on an Agilent SB-C18 column (4.6 mm×150 mm, 5 μm) by gradient elution using acetonitrile and water-acetic acid(100∶0.05, v/v) as the mobile phase. Seven major components in RA were successfully separated. This quantitative method was fully validated in respect of the following performance criteria: linearity, precision, repeatability, stability, accuracy, limits of detection (LOD) and quantification (LOQ). A formula database of known compounds in RA was established, against which, most of the reported components in this herbal extract were identified effectively based on the extract masses acquired by TOFMS. This qualitative and quantitative method was successfully used to analyze the components in 10 batches of RA samples collected from different regions in China. This global quality control method, which consisted of HPLC-DAD-MS assay of seven major components and unambiguous identification of nineteen components, is suitable for routine quantification and comprehensive quality control of RA.

Rapid determination of ferulic acid and identification of its structure in rhizome of Phragmites communis by GC-TOFMS / 中草药

Objective To make a rapid determination of ferulic acid and identification of its structure in rhizome of Phragmites communis (RPC). Methods The chemical constituents were extracted by solvent and the relative content and structure of ferulic acid in RPC were determined and identified using GC-TOFMS technique. Results By GC-TOFMS, the ferulic acid was successfully isolated from the extracts of RPC in a relative content of 5%. According to the exact masses and corresponding elemental compositions of molecular ion and eight characteristic ions, a possible fragmentation pathway of the ferulic acid molecule was proposed, by which the molecular structure was confirmed. Conclusion GC-TOFMS technique is a rapid, accurate, sensitive, and reliable method for determination of the relative content of ferulic acid in RPC and identificatiion of its structure.