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Semen of Cuscuta chinensis has been reported to have an anti-osteoporosis effect, however, the components which account for the anti-osteoporosis effect have not been clarified. In this work we propose a biochemometrics strategy that integrates quantitation, anti-osteoporosis evaluation in zebrafish, and grey relationship analysis for the identification of anti-osteoporosis components from the semen of Cuscuta chinensis. In the beginning, a precise and accurate liquid chromatography-tandem mass spectrometry method was established for simultaneous quantitation of seven major components in crude and salt-processed Cuscuta chinensis. The mode of multiple reaction monitoring was used. Chloramphenicol was selected as the internal standard. The method showed good linearity and repeatability. The recovery rates of each component ranged from 95.4 to 103.9%. The precisions of intra-day and inter-day were all within 5.0%. The method was then applied for quantitation of the seven major components in 11 batches of crude and salt-processed Cuscuta chinensis. Subsequently, the anti-osteoporosis effects of crude and salt-processed Cuscuta chinensis were evaluated in zebrafish. Principle component analysis, grey relationship analysis, and partial least squares regression were applied for deciphering the relationship between the contents of seven major components and the anti-osteoporosis effects. Hyperin, p-hydroxycinnamic acid, and astragalin were found to be the major anti-osteoporosis components.

A novel quality evaluation method of Corydalis yanhusuo was established by researching the high-performance liquid chromatography behavior of alkaloids under different buffer solutions and exploring the correlation between alkaloids in C. yanhusuo. The retention times of tetrahydropalmatine and corydaline were significantly influenced by pH, while the peak shape was affected by buffer types and ionic strength. The resolution of compounds in fingerprint was satisfactory under acetonitrile-0.2% phosphoric acid buffer (adjusted pH to 5.0 with triethylamine). Twelve common peaks were found by comparing 20 batches of C. yanhusuo fingerprints, and three tertiary alkaloids and four quaternary alkaloids were identified. The fingerprints were analyzed by similarity analysis, hierarchical cluster analysis, principal component analysis, and partial least squares discriminant analysis. All samples were divided into three groups, and the contents of dehydrocorydaline and coptisine from Zhejiang province were relatively higher than other origins. There were six components performing more contributions to the quality of C. yanhusuo. The correlations between alkaloids were conducted by Pearson correlation analysis and mathematical model analysis. The content correlation between palmatine and berberine was y = 0.28x2 + 0.03x + 0.03, and the dehydrocorydaline and coptisine was y = -7.54/(1 + (x/0.14)0.5 ) + 2.61. The established mathematical model of alkaloids provided a guiding significance for the quality control of C. yanhusuo.

A DDQ-mediated tandem reaction of easily available o-hydroxy­aryl enaminones and cycloheptatriene is disclosed. It provides an effective and convenient method for the synthesis of 3-(cycloheptatrienyl)-substituted chromones in moderate to excellent yields with good functional group tolerance; this reaction involves an oxidative coupling and intramolecular annulation pathway.

Background Honokiol and magnolol were considered as markers for the analysis of Cortex Magnoliae Officinalis, its related Chinese Patent Medicines and their metabolites. However, the determination of these two analytes in a water-soluble sample is difficult and therefore requires a more efficient method. Objective To develop a sensitive method for the determination of honokiol and magnolol in a water-soluble sample for better quality control of Cortex Magnoliae Officinalis and its related Chinese Patent Medicines. Method In this work, a combination of dispersive micro-solid-phase extraction (DMSPE) and high-performance liquid chromatography (HPLC) has been developed for simultaneous preconcentration and determination of honokiol and magnolol in complex bio-samples. Several experimental factors affecting the extraction efficiency were optimized by single factor test. Results Under the optimized extraction conditions, the proposed method exhibited good linearity of not less than 0.9998, satisfactory precision with relative standard deviation of less than 1.3%, and acceptable mean recoveries of 97.3% and 101.5% for honokiol and magnolol, respectively. Furthermore, the method exhibits extremely high sensitivity with detection limits of 0.0097 and 0.0231 ng/mL, which is even more sensitive than those methods developed by MS. Conclusions The method established in this study is fast, economic, accurate, easy to operate, and importantly well suited to the extraction and analysis of honokiol and magnolol in a real complex sample matrix.

An oxidative annulation for the synthesis of 2,4-diarylquinolines from o-allylanilines is disclosed that uses recyclable reagent Chloranil­ as the oxidant. The corresponding products are obtained in moderate to excellent yields. Furthermore, a one-pot access to 2,4-di­aryl­quinolines from easily available anilines and 1,3-diarylpropenes is described as a highly atom-efficient protocol that involves oxidative coupling, rearrangement, and oxidative annulation.

Four stereoisomeric components were produced during the synthesis of the antidepressant drug (1S, 4S)-sertraline hydrochloride due to the two chiral carbon centers in its chemical structure, including (1S, 4S), (1R, 4R), (1S, 4R), and (1R, 4S)-isomer. Stereoselective separation of the target isomer (1S, 4S)-sertraline from the medicinal reaction mixtures by countercurrent chromatography using hydroxypropyl-β-cyclodextrin as the stereoselective selector was investigated. A biphasic solvent system composed of n-hexane/0.20 mol/L phosphate buffer solution with pH 7.6 containing 0.10 mol/L of hydroxypropyl-β-cyclodextrin (1:1, v/v) was selected for separation of cis-sertraline and trans-sertraline using reverse phase elution mode and (1S, 4S)-sertraline was separated with (1R, 4R)-sertraline using recycling elution mode. A fabricated in-house analytical countercurrent chromatographic apparatus was used for optimization of the separation conditions. Stationary phase retention and peak resolution were investigated for separation of cis-sertraline and trans-sertraline by the analytical apparatus.

Introduction Germacrone, furanodiene and β-elemene are the representative bioactive compounds in Curcuma species. The conventional extraction methods of these three sesquiterpenes are usually time-consuming and require a large volume of hazardous organic solvents. Thus, a fast and reliable method for extracting these sesquiterpenes from Curcuma plant is required urgently. Objective To establish a novel and simple extraction method for quantitative analysis of small amounts of sesquiterpenes in C. wenyujin plant. Methodology A method using microwave-assisted ionic liquid-micelle extraction combined with dispersive micro-solid-phase extraction (DMSPE) has been proposed for the extraction of three sesquiterpenoids in Curcuma plant. Fluorinated carbon nanotubes (FCNTs) were used as an adsorbent in DMSPE for the first time. Parameters concerning the microwave-assisted extraction (MAE) conditions and the DMSPE were investigated and evaluated to achieve optimum extraction efficiency of target analytes. Results The final conditions of ionic liquid-micelle based MAE were selected to be 0.25 M of 1-decyl-3-vinylimidazolium bromide as the extraction solvent, microwave irradiation for 10 min at 60°C. And the optimal DMSPE conditions were found to be 2 μg/mL of FCNTs as the adsorbent, extraction time of 2 min and 100 μL of acetonitrile as the elution solvent. The developed method exhibited good linearities (R > 0.9990), high repeatability and recoveries. The proposed method has been successfully applied in determination of sesquiterpenes in C. wenyujin samples. Conclusion The work shows a potential in analysing small amounts of sesquiterpenes in complex samples and represents the first attempt of using FCNTs as an adsorbent for the microextraction mode.

A sensitive dispersive micro solid-phase extraction coupled with HPLC has been developed for preconcentration and determination of three flavonoids (quercetin, kaempferol, and isorhamnetin) in complex matrix samples. Parameters that affect extraction efficiency have been optimized. The optimal extraction conditions are using 2 μg/mL of crab shell as the sorbent, extraction for 2 min at pH 7, and then eluting with 100 μL of methanol. As a result, the method shows good linearity (R > 0.9994), low LODs (even 0.08 ng/ml) and satisfactory recovery in real honey and rat urine samples. As an eco-friendly biomaterial, crab shell powder is used as sorbent in pretreatment of flavonoids, and its adsorption mechanism has been investigated for the first time. Compared with the other reported methods, the proposed strategy is time-saving, eco-friendly, and highly sensitive using HPLC (even achieving MS grade sensitivity).

To avoid stationary phase irreversibly adsorption and tedious and time-consuming separation steps, high-speed counter-current chromatography was employed for the preparative separation of anti-tumor compound antroquinonol from solid fermentation culture of Antrodia camphorata for the first time. A Box-Behnken experimental design, based on three parameters including liquid-to-solid ratio, extraction time and extraction temperature, was applied to optimize the ultrasonic extraction procedure. The optimal extraction condition was set as below: liquid-to-solid ratio: 49.57:1; extraction time: 55.76 min; extraction temperature was arranged as 44.21 °C. Meanwhile, an optimized solvent system containing petroleum ether, ethyl acetate, methanol and water (4:1:4:1, v/v/v/v) was selected for the preparative separation of antroquinonol at a flow rate of 2.0 mL/min. The yield of isolated antroquinonol was determined to be 6.0 mg from 0.67 g of ethyl acetate extracts. The isolated antroquinonol was elucidated by ultra high performance liquid chromatography tandem mass spectrometry, and NMR spectroscopy, and by comparison with literature data. The purity of isolated antroquinonol was determined to be 97.12%. This study confirmed high-speed counter-current chromatography was powerful and cost-effective for the preparative separation of the high-potently anti-tumor compound antroquinonol from solid fermentation culture of Antrodia camphorata. This article is protected by copyright. All rights reserved.

The analysis of anionic compounds via the sensitive FLM-EKS method remains challenging due to their inverse migration behaviors to EOF in the capillary. A highly sensitive ionic liquid coating-based online preconcentration methodology via electrokinetic supercharging (EKS) coupled to a free liquid membrane (FLM) was first proposed and successfully used to determine vanillic acid (VA), salvianic acid A sodium (SAAS) and ferulic acid (FA) in a complex sample matrix. EKS was recognized as the first step preconcentration, and its separation conditions were optimized. Then, the second preconcentration step, namely FLM, which was performed by simple injection of a short water-immiscible solvent plug as a membrane interface prior to electrokinetic injection (EKI) of the sample, was further investigated to increase the stacking efficiency. The influences of multifarious parameters on FLM-EKS performance were examined, and the optimal FLM-EKS conditions were identified: a buffer electrolyte of 20 mM Tris-HOAc (pH=6) with 5% (v/v) methanol; hydrodynamic injection (HI) of a leading electrolyte (50 mM NH4Cl, 30 s, 0.5 psi) followed by the HI of 2-nitrophenyl octyl ether (NPOE) as the FLM (5 s, 0.5 psi); EKI of the sample (120 s, -10 kV) and there with the HI of terminator (20 mM N-cyclohexyltaurine, CHES, 40 s, 0.5 psi). The newly developed method yielded analytes with 2006-2862-fold enhancement compared to the normal injection, there by producing detection limits of 1.5-5.0 ng mL-1 with satisfactory repeatability below 2.92%, which provides 1.5-2.0 times higher stacking efficiency than that obtained by EKS only. Feasibility in real sample analysis was demonstrated by using Naoxintong capsules and rat feces, which suggests that the presented methodology has a high potential for the reliable and sensitive determination of trace anionic components in complex sample matrices.

Background: Shenqi Jiangtang Granule (SJG), a classical prescription of traditional Chinese medicine, is widely used to treat diabetes and its complications. Although, the clinical efficacy of SJG, is sufficient, the pharmacokinetic behavior of various substances in the plasma of SJG is unknown. Objective: The aim of this study was to investigate the plasma pharmacokinetics during absorption of SJG after oral administration in rats. Methods: A rapid and accurate ultra-high performance liquid chromatography/tandem mass spectrometry (UPLC- MS/MS) method was developed for the simultaneous determination of eight analytes in SJG, including gomisin D, schisandrin A, schisandrin B, schizandrol A, schizandrol B, ginsenoside Rd, ginsenoside Re and notoginsenoside Ft1. The analysis was carried out on a BEH C18 column (2.1 mm × 50 mm, 1.7 μm) with gradient elution at a flow rate of 0.2 mL/min in a mobile phase consisting of 0.1% formic acid water and acetonitrile. In addition, lignans and saponins were detected in positive ion mode and negative ion mode, respectively. Results: Eight analytes in SJG, including gomisin D, schisandrin A, schisandrin B, schizandrol A, schizandrol B, ginsenoside Rd, ginsenoside Re and notoginsenoside Ft1, showed good linearity (R2 in the range of 0.9955 ~ 0.9999). The lower limit of quantification (LLOQ) was 5, 0.8, 0.8, 8, 0.8, 5, 0.6 and 10 ng/mL. The accuracy and precision of all analytes were at ±15%. Matrix effect and average extraction recovery were > 85%. All analytes performed well under four storage conditions. Conclusions: The results showed that in vivo absorption and exposure of gomisin D and ginsenoside Rd were better than other analytes, while schizandrol B and notoginsenoside Ft1 were poorly absorbed. This approach could be applied to study the pharmacokinetic characteristics of various analytes in plasma after oral administration of SJG in rats.

Electrokinetic supercharging, a convenient and powerful online preconcentration technique in capillary electrophoresis, was introduced and evaluated for the determination of two alkaloids, berberine and jatrorrhizine, in mice fecal samples for the first time. The method depended on using a bare fused silica capillary (50 cm × 50 μm i.d.) and applying the voltage of 25 kV with UV detection at 205 nm. Parameters that affect the separation and preconcentration efficiency have been optimized. The optimum conditions used were as follows: background electrolyte consisting of 40mM sodium dihydrogenphosphate containing 30% methanol (v/v); hydrodynamic injection of 20mM KCl (50 mbar × 150 s) as the leading electrolyte; electrokinetic injection of the sample (+15 kV, 120 s) followed by the hydrodynamic injection of 30mM dodecyl trimethyl ammonium chloride (50 mbar × 12 s) as the terminating electrolyte. The results showed that the detection sensitivity of berberine and jatrorrhizine was, respectively, improved up 2740- and 2928-fold compared with normal injection, providing limits of detection lower than 3 ng/mL with good repeatability in areas (relative standard deviation < 3%). In summary, the developed method proved its ability in analyzing trace alkaloids in complicated biological samples.

A rapid and sensitive UPLC-MS/MS method was established for the simultaneous quantification of 6'''-feruloylspinosin, spinosin, jujuboside A, and jujuboside B in rat plasma after the oral administration of traditional and dispensing granules (DG) decoction of Ziziphi Spinosae Semen (ZSS). The four components were separated using 0.1% formic acid and acetonitrile as a mobile phase by gradient elution at a flow rate of 0.3 mL/min equipped with a C18 column (2.1 × 50 mm, 1.7 μm particle size, Acquity BEH C18 ). The mass spectrometer was operated under multiple reaction monitoring mode. An aliquot of 100 μL rat plasma was deproteinized by 300 μL methanol. The supernatant was injected into the UPLC-MS/MS system for analysis. The calibration curves displayed good linearity. The intra-day and inter-day precisions (RSD) were less than 7.3%. The accuracies ranged from -1.3 to 6.1%. The extraction recoveries ranged from 95.8 to 101.9%, and the matrix effects were satisfactory. For DG, half-life values (t1/2 ) of 6'''-feruloylspinosin and Cmax of jujuboside A were elevated remarkably. MRT0-t of jujuboside B was significantly increased. No significant variation was observed for the pharmacokinetic parameters of spinosin. The results could provide a scientific basis for the clinical application of traditional and DG decoction of ZSS.

Three key intermediates in the preparation of the nonsteroidal anti-inflammatory drug naproxen were successfully separated and purified with high purity from synthetic mixtures by countercurrent chromatography with a selected biphasic solvent system. The biphasic solvent system composed of n-hexane/ethyl acetate/methanol/water (9:1:9:1, v/v/v/v) was selected according to partition performance of the three components using thin-layer chromatography. Fifty milligrams of the synthetic mixture after the three-step reaction was injected into a preparative countercurrent chromatography separation column and yielded 3.5, 14.0, and 8.0 mg of three key intermediates with 95.0, 99.0, and 98.0% purity, and the recovery of each component was 65.2, 71.2, and 69.6%, respectively. The results indicated that countercurrent chromatography is an efficient alternative and economical method for the separation and purification of intermediate components from synthetic mixtures.

A simple, efficient and environmental friendly method was proposed for determining five sesquiterpenoids of Curcuma wenyujin by MSPD extraction coupled with MEEKC separation. Molecular sieve was applied as a solid support for extraction of sesquiterpenoids for the first time. Various parameters affecting extraction and separation efficiency were investigated. The optimized conditions involved dispersing sample (200 mg) with 200 mg of TS-1 for 150 s and using 1000 μL of methanol to elute five target analytes. Finally, they were well separated by using a running buffer containing 1.3% SDS, 5.0% 1-butanol, 0.5% ethyl acetate and 10% acetonitrile in 10 mM borate buffer at pH 9.0. Consequently, the developed method was fully validated and successfully applied to determine the five sesquiterpenoids including curdine, curcumenol, germacrone, furanodiene and β-elemene in Curcuma wenyujin origin's Chinese herbal medicines. Furthermore, hierarchical cluster analysis was performed based on the contents of target compounds for distinguishing steamed and non-steamed drugs. The present study provided a promising method for fast investigation and discrimination of chemical difference in steam & non-steamed Chinese medicines from Curcuma wenyujin origin.

A novel, metal-free cross-dehydrogenative coupling (CDC) reaction of C(sp2)–H bonds of enamines and α-oxo ketene dithioacetals with C(sp3)–H bonds of 1,3-diarylpropenes mediated by 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) is reported. The α-alkylation products are obtained with moderate to good yields. The method provides an efficient and alternative strategy for the synthesis of the corresponding products.

Background Due to the complexity of traditional Chinese medicine (TCM), the current quality evaluation of TCM are difficult to associate with clinical efficacy. Shenqi Jiangtang Granule (SJG), a classical TCM formula, is proven as a therapy for treatment of type II diabetes mellitus (DM) and complications while the substantial basis of the therapeutic effects is not clear. Purpose The present study proposed an integrated approach to discriminate the quality markers (Q-markers) based on multi-dimensional characteristic network for quality control of TCM. Methods The multi-dimensional characteristic network was established by “Spider-web” mode, which was comprehensively integrating “compatibility-content-activity- efficiency-stability” of the candidate ingredients. The activity dimension was evaluated by the inhibitory activity of SJG on α-glucosidase and aldose reductase. The efficacy dimension was assessed through the association between the compounds and the target pathway of diabetic nephropathy (DN) based on integrated pharmacology platform. Each dimension for the feature network was quantified by multivariate statistical analysis, and regression area of the candidate compounds was constructed in the network. Finally, the candidate compounds were sorted comprehensively by the regression area. Results A total of 30 chemical compounds with effective hypoglycemic activity were identified as the potential Q-markers. From the data analysis, three dimensions of activity, efficacy and content performed a greater impact on the regression area of the characteristic network. Among these compounds, ginsenoside Re, ginsenoside Rd, ginsenoside Rg1, calycosin, ginsenoside Rb1, formononetin, astragaloside IV, ginsenoside Rf, ginsenoside Rc, notoginsenoside Fe, schisandrol A, gomisin D were screened out as the candidate Q-markers of SJG. Conclusion The multi-dimensional characteristic network integrating compatibility, content, activity, efficiency and stability is efficient to discriminate the potential Q-markers of TCM prescription. Our results demonstrated that 12 candidate compounds from Panax Ginseng, Radix Astragali and Schisandrae Chinensis might select as Q-markers for qualitative evaluation of SJG.

3-Phenyllactic acid is an antimicrobial compound with broad-spectrum activity against various bacteria and fungus. The observed difference in pharmacological activity between optical isomeric 3-phenyllactic acid necessitates a method for enantioseparation. Chiral ligand exchange countercurrent chromatography was investigated for the enantioseparation of 3-phenyllactic acid with a synthesized chiral ligand. A two-phase solvent system was composed of n-butanol/hexane/water (0.4:0.6:1, v/v/v) to which N-n-dodecyl-l-hydroxyproline was added to the organic phase as chiral ligand and cupric acetate was added in the aqueous phase as a transitional metal ion. The influence factors were optimized by enantioselective liquid-liquid extraction. Baseline enantioseparation of racemic 3-phenyllactic acid by analytical high-speed countercurrent chromatography was achieved. The optical purities of enantiomeric 3-phenyllactic acid reached 99.0%, as determined by chiral high-performance liquid chromatography.

The metal-free cross-dehydrogenative coupling (CDC) reaction and subsequent cyclization of 2-hydroxy-1,4-naphthoquinone (lawsone) and 1,3-diarylpropene promoted by DDQ has been developed. 2-Hydroxy-3-alkyl-1,4-naphthoquinones and pyranonaphthoquinones with potential pharmaceutical applications are obtained in moderate to good yields. The reaction is also compatible for 4-hydroxycoumarins.

A plasma-generated carbon aerosol, with the merits of high surface area, abundant porosity and high crystallinity, is a mass synthesized by-product from direct methane conversion in the production of syngas or hydrogen. In the present work, a carbon aerosol produced by rotating gliding arc (RGA) plasma was firstly incorporated with a commercial sponge to fabricate a highly selective and hydrophobic adsorption material. The properties of the carbon aerosol and its derived sponge were comprehensively characterized. By a simple dip-coating method, the wettability of the pristine sponge was altered, which was attributed to the plasma coating of the carbon aerosol. The modified sponge demonstrated an effective and selective adsorption ability for a wide range of oils and solvents, with the maximum adsorption capacity of up to 86 times its own weight. Moreover, the used sponge could be easily recovered by simple evaporation or manual squeezing, while maintaining approximately 100% of its starting adsorption capacity over 5 adsorption-recovery cycles. Such a plasma carbon aerosol coated sponge exhibits a great prospect as a cost-efficient, recyclable and scalable material in separation and adsorption for water treatment.

We proposed a biochemometrics strategy for tracing diuretic components of herbs based on quantitative determination and pharmacological evaluation. First, a sensitive and robust liquid chromatography coupled with tandem mass spectrometry approach was established for simultaneous quantification of six major triterpenoids in crude and salt-processed Alisma orientale. The separation of triterpenoids was achieved on a BEH C18 column with a mobile phase consisting of acetonitrile and water spiked with 0.1% formic acid. Six major triterpenoids were detected by multiple reaction monitoring in the negative ion mode. Glycyrrhetinic acid was used as the internal standard. The approach showed good linearity. Intra- and inter-day precisions were all within 2.9%. The recovery rates of each triterpenoid ranged from 97.9% to 103.2%. The approach was then successfully employed for quantitative analysis of six triterpenoids in ten batches of crude and salt-processed A. orientale. Second, the diuretic effects of crude and salt-processed A. orientale were evaluated in mice. Third, principal component analysis and canonical correlation analysis were used to uncover the relationship between the contents of six major triterpenoids and the diuretic effect of different crude and salt-processed samples. Alisol B, alisol F, and alisol A have a close positive correlation with the diuretic effect.

A simple and reliable liquid chromatography-mass spectrometry (LC-MS) method was developed for simultaneous determination of saikosaponin A, saikosaponin B1 , saikosaponin C, saikosaponin D and saikosaponin F in rat plasma using glycyrrhetinic acid as an internal standard (IS). The separation was operated on a Waters BEH C18 column. The mobile phases of gradient elution consisted of acetonitrile (A) and 0.1% aqueous acetic acid (B). The mass spectrometric detection was accomplished in multiple reaction monitoring mode. The five saponins displayed good linearity (r2 > 0.9996). The lower limits of quantitation of saikosaponin A, saikosaponin B1 , saikosaponin C, saikosaponin D and saikosaponin F were determined to be 2.9, 2.3, 3.5, 2.9 and 3.1 ng/mL, respectively. Moreover, the intra- and inter-day precisions of the five saponins showed an RSD within 2.96%, whereas the accuracy (RE) ranged from -2.28 to 2.78%. Finally, the developed method was fully validated and applied to a comparative pharmacokinetic study of the five bioactive saponins in rats following oral administration of crude and vinegar-processed Bupleurum scorzonerifolium.

A novel on-line synergistic proconcentration strategy coupling field-amplified sample stacking and micelle to cyclodextrin stacking for cationic analytes in capillary zone electrophoresis has been proposed and applied for the separation and determination of two alkaloids, matrine, and oxymatrine in complicated matrix samples. The approach was performed by the long injection of sample in a low-conductivity sodium dodecyl benzene sulfonate solution followed by the injection of hydroxypropyl-β-cyclodextrin solution in higher conductivity. The stacking mechanism of this method has been expounded and parameters affecting stacking effect have been optimized in our study. Under the optimum experimental conditions, 169- and 218-fold sensitivity improvements were achieved for matrine and oxymatrine when compared with normal injection. Analytical indicators including linearity, limits of detection, and reproducibility (intra- and inter-day relative standard deviations) were evaluated. Moreover, sample matrix effect was studied using compound flavescent sophora and salicylic acid powder and spiked urine samples. The developed method is an attempt for the combination of micelle to cyclodextrin stacking with other stacking methods. It could be a good alternative choice for the determination of alkaloids in a complex sample matrix.

Corydalis turtschaninovii (CT) is a traditional Chinese medicine which is known to have analgesic effects, and is under investigation for the management of chronic pain. Our study aims to establish a UPLC–MS/MS method for pharmacokinetic determination of nine bioactive alkaloids of raw and processed CT in rat plasma. Nitidine chloride was selected as internal standard. After protein precipitation with methanol, the plasma samples were separated on a reversed phased column with a mobile phase of acetonitrile and water (including 0.1% formic acid). The MRM parameters were optimized as follows: m/z 354.0 → 188.1 for protopine, m/z 321.0 → 293.1 for coptisine, m/z 371.1 → 189.1 for allocryptopine, m/z 357.1 → 193.3 for tetrahydropalmatine, m/z 324.1 → 176.1 for tetrahydrocoptisine, m/z 340.1 → 176.2 for tetrahydroberberine, m/z 368.1 → 289.1 for corynoline, m/z 370.5 → 192.1 for corydaline, m/z 367.2 → 351.2 for dehydrocorydaline, and m/z 406.0 → 300.3 for the IS. The linearity, accuracy, precision, stability, recovery and matrix effect of the method were well validated. This method was successfully employed for a pharmacokinetic study of raw and vinegar-processed CT in rats. The absorption of the nine alkaloids was accomplished in an hour. The double peak phenomenon of the nine alkaloids may be ascribed to enterohepatic recirculation. Compared with the raw group, AUC0→t and Cmax of the nine alkaloids were significantly elevated in the vinegar-processed group. Our findings suggest that vinegar-processing could increase the bioavailability of the nine alkaloids of CT in rats. The pharmacokinetic information obtained will provide basis for application of processed CT in future clinical therapy.

Alisma plantago-aquatica is known to regulate water and fluid balance in cells, and is under testing for the therapy for patients suffering from chronic nephritis. Herein, a UHPLC-MS/MS method was established and validated for the determination of six bioactive triterpenoids of raw and salt-processed Alisma plantago-aquatica in rat plasma. The acquired plasma was subjected to protein precipitation with acetonitrile. Glycyrrhetinic acid was employed as internal standard. The pretreated samples were separated on a reversed phased column with a mobile phase of acetonitrile and water (including 0.1% formic acid). The MRM mode for the six triterpenoids were at m/z 535.4 → 489.4 for alisol A, m/z 517.3 → 471.4 for alisol B, m/z 533.3 → 487.3 for alisol F, m/z 577.4 → 531.4 for alisol A-24-acetate, m/z 559.4 → 495.4 for alisol B-23-acetate, m/z 573.3 → 509.3 for alisol C-23-acetate, and 469.3 → 425.3 for the IS. The accuracy and precision of the method were determined as −2.2%-3.6% and 0.8%–3.0%, respectively. This approach was employed to a pharmacokinetic study of the six bioactive triterpenoids after intragastric administration of raw and processed Alisma plantago-aquatica in rats. The two-phasic pharmacokinetic of alisol B, alisol C-23-acetate and alisol F were reported for the first time, which may be ascribed to enterohepatic recirculation of these triterpenoids.

A simple and efficient ultra-miniaturized pipette-tip matrix solid-phase dispersion microextraction using molecular sieve as adsorbent has been developed to simultaneously determine honokiol, magnolol, costunlide and dehydrocostus lactone in a complex Chinese medicine. The types of adsorbent and elution solventswere optimized by single factor experiment. Adsorbent/sample ratio, grinding time and elution volume were optimized by Box-Behnken design (BBD). The optimum extraction is: using SBA-3 as an adsorbent, 200 μL of methanol as elution solvent, 2:1 of sample/adsorbent ratio and 130 s of grinding time. The proposed method applied in real samples provides good linearity (R2 > 0.9999), sufficient accuracy, high reproducibility (RSD

Ethnopharmacological relevance Shenqi Jiangtang granule (SJG) is an ancient Chinese herbal formula used for treatment of Diabetes mellitus and its complications. Aim of the study To establish an integrated approach for discovery of effective Aldose reductase inhibitors (ARIs) from SJG. Materials and methods An integrated approach combining ultrafiltration-liquid chromatography-mass spectrometry (UF-LC-MS) with in silico molecular docking was established for development of ARIs. AR enzyme was separated from the rabbit's crystalline lens. The inhibitory activities of these compounds were detected by UV spectrophotometry with DL-glyceraldehyde as a substrate. Furthermore, molecular docking was used to understand the binding mechanism of these screened compounds interacting with AR. Results After optimization of AR reaction system and ultrafiltration incubation system, 17 active ingredients were screened from SJG by UF-LC-MS technique. Among these potential AR inhibitors, ginsenoside Rd exhibited the strongest activity with IC50 value of 45.77 μM. Three of them, calycosin, gomisin J and schisandrin A were demonstrated to be potential inhibitors for the first time, with IC50 at 447.34 μM, 181.73 μM, and 429.00 μM, respectively. Most of the active compounds exhibited competitive inhibition against AR. The docking scores of saponins were higher than that of lignans, which was consistent with the verification results. Conclusion The results indicated that TCM formula with clinical efficacy was indeed hopeful source for screening active ingredients, and the combination of UF-LC-MS and in silico molecular docking was a universal and promising approach for development of effective enzyme inhibitors.

Selective separation of chemical components from seven kinds of volatile oil by countercurrent chromatography with three types of cyclodextrins as selective reagent was investigated in this work. Preparative separation of chemical components from volatile oil is generally quite challenging due to their extremely complexity of the composition. A biphasic solvent system n-hexane-0.10 mol L(-1) cyclodextrin (1:1, v/v) was selected for separation of components from volatile oil and three types of cyclodextrins were investigated, including β-cyclodexrin, methyl-β-cyclodexrin and hydroxypropyl-β-cyclodexrin. All kinds of volatile oils are from seven kinds of traditional Chinese herb. Results showed that some chemical components could be well separated with high purity from each kind of volatile oil using different type of cyclodextrin as selective reagent. For example, germacrone and curcumenone could be selectively separated from volatile oil of Curcumae Rhizoma with methyl-β-cyclodexrin and hydroxypropyl-β-cyclodexrin as selector respectively, and other five components were selectively separated from volatile oil of Chuanxiong Rhizoma, Myristicae Semen, Aucklandiae Radix and Angelicae Sinensis Radix by countercurrent chromatography with different cyclodexrin as selective reagent. Separation mechanism for separation of components from volatile oil by countercurrent chromatography with cyclodextrin as selective reagent was proposed. Peak resolution of the present separation method could be greatly influenced by the chemical compositions of volatile oil.

Catalyzed by 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) and sodium nitrite, the coupling reaction of 1,3-diarylpropynes and 1,3-dicarbonyl compounds using molecular oxygen as a terminal oxidant was developed. The reaction was promoted dramatically in the presence of HCOOH. The corresponding products were obtained within half an hour in 37–87% yields.

Sertraline is an antidepressant in a group of drugs called selective serotonin reuptake inhibitors. Four stereoisomeric compounds would be produced in its synthetic preparation due to two chiral carbons on its chemical structures. In the present work, stereoselective liquid-liquid extraction of isomeric sertraline with substituted cyclodextrins as stereoselective extractant was investigated. Factors affecting the distribution performance, including organic solvents, types of extractants, pH value, buffer solution of aqueous phase, concentration of extractant and temperature, were investigated. Under optimized conditions, a stereoselectivity of 1.404 was obtained for cis-sertraline and a stereoselectivity of 2.373 was obtained for trans-sertraline when hydroxypropyl-β-cyclodextrin was used as the stereoselective extractant, and a stereoselectivity of 1.685 was achieved for trans-sertraline with methyl-β-cyclodextrin as extractant. An unusual stereoselective combination was observed for trans-sertraline and cis-sertraline when sodium carbonate buffer was used. Successful stereoselective separation of trans-sertraline and cis-sertraline, (1S, 4R) and (1R, 4S)-sertraline by analytical countercurrent chromatography with methyl-β-cyclodextrin as stereoselective selector was achieved, using a biphasic solvent system composed of n-hexane : 0.1 mol L−1 citrate buffer solution with pH7.6 (1:1, v/v).

Four polysaccharide fractions (P-1: 71.40%, P-2: 1.95%, P-3: 1.14%, P-4: 1.64%) were isolated from crude Polygonatum sibiricum polysaccharide (PSP), processed by water extraction, ethanol precipitation, and further separated with diethylaminoethyl cellulose-52 anion-exchange chromatography. Their molecular weights and monosaccharide compositions were characterized by high performance gel chromatography with evaporative light scattering detector and ultraviolet-visible detector. The antioxidant activity of four polysaccharides fractions were assessed by the electron transfer menchanism (DPPH, ferric reducing power, and ABST assays) and chelation of transition metals (Fe2+ and Cu2+ chelation ability). The highest content fraction P-1 exhibited the lowest antioxidant activity, and the ranking of antioxidant capacity was P-4 > P-3 > P-2 > PSP > P-1. After processed by microwave-assisted degradation, the molecular weight of P-1 was decreased from 2.99 × 105 to 2.33 × 103 Da, while the antioxidant activity of degraded P-1 was about eightfold higher than natural P-1. These results indicated that the proposed microwave-assisted degradation approach was an efficacious methodology to improve their bioactivity by lower the molecular weight of polysaccharides. PRACTICAL APPLICATION: This study provided an environmentally friendly, convenient and efficient microwave-assisted degradation technology to process the neutral polysaccharides from Polygonatum sibiricum. The results could be used for the development and utilization of various plant polysaccharides as a kind of food supplement in our daily life.

A sensitive, simple, and environmentally friendly online three-step stacking strategy combining field-enhanced sample injection, sweeping and micelle to solvent stacking has been developed to determine trace amounts of organic anionic analytes in complex biological samples. A green ionic liquid, 1-dodecyl-3-methylimidazolium hydrogen sulfate, was first introduced as a micellar solution and used for electroosmotic flow reversal in this stacking strategy. The mechanism of stacking has been discussed, and parameters affecting three-step stacking preconcentration efficiency have been optimized. The capillary coated with an ionic liquid was easy to prepare, regenerable and repeatable (RSD

Racemic trans-paroxol, trans-N-methylparoxetine and trans-paroxetine containing two chiral centres were stereoselectively separated using countercurrent chromatography with hydroxypropyl-β-cyclodextrin as the chiral selector. A two-phase solvent system composed of n-butyl acetate and 0.1 mol L−1 sodium carbonate-sodium bicarbonate buffer at pH 9.2 (1:1, v/v) was selected, and 0.10 mol L−1 hydroxypropyl-β-cyclodextrin was added to the aqueous phase as the chiral selector. Racemic trans-N-methylparoxetine and racemic trans-paroxol (20 mg of each) were stereoselectively separated by countercurrent chromatography in an individual run, yielding 7.1–8.3 mg of enantiomers with a purity of 95–98%, where the recovery of each separated isomer reached approximately 70–83%. Racemic trans-paroxetine (20 mg) was stereoselectively separated by countercurrent chromatography using a recycling elution mode with a biphasic solvent system composed of n-hexane: n-butyl acetate: 0.1 mol L−1 sodium carbonate-sodium bicarbonate buffer at pH 9.2 (9:1:10, v/v/v), and 0.10 mol L−1 hydroxypropyl-β-cyclodextrin was added to the aqueous phase as the chiral selector, yielding 5.0–5.6 mg of enantiomer with a high purity of over 98–99%.

Objective: To establish a quantitative analysis of multi-components by single marker (QAMS) method for the determination of triterpenoids and distinction of Ganoderma and related species. Methods: Ten ganoderma triterpenoids were determined by HPLC in addition to a QAMS method. On the basis of QAMS, principal component analysis (PCA) and cluster analysis (CA) were used to distinguish Ganoderma samples. Results: Ten types of ganoderma triterpenoids had good linearity of regression coefficients (R2 ≥ 0.9992). Relative correction factors (RCFs) of lucideric acid C, lucideric acid LM1, ganoderic acid G, lucideric acid B, lucideric acid A, ganoderenic acid D, ganoderic acid D, lucideric acid D and ganoderic acid F with reference to ganoderic acid A were 0.929, 1.011, 0.608, 1.041, 0.997, 2.464, 0.891, 0.826 and 0.722, respectively. Ten types of ganoderma triterpenoids in twenty-nine batches of Ganoderma and related species were determined by QAMS. Nineteen batches of samples containing ganoderma acid A were roughly divided into three categories by PCA and CA. Conclusion: The developed QAMS method, in combination with PCA and CA, served as an accurate method for the rapid evaluation and authenticity identification of Ganoderma.

In this work, a simple and efficient protocol for the rapid separation of two pairs of isomeric monoterpenes from Paeoniae Alba Radix was developed by combining macroporous resin and elution-extrusion counter-current chromatography. The crude extract was firstly subjected to a D101 macroporous resin column eluted with water and a series of different concentrations of ethanol. Then, effluents of 30 and 95% ethanol were collected as sample 1 and sample 2 for further counter-current chromatography purification. Finally, a pair of isomers, 96 mg of compound 1 and 48 mg of compound 2 with purities of 91.1 and 96.2%, respectively, was isolated from 200 mg of sample 1. The other pair of isomers, 14 mg of compound 3 and 8 mg of compound 4 with purities of 93.6 and 88.9%, respectively, was isolated from 48 mg of sample 2. Their purities were analyzed by high-performance liquid chromatography, and their chemical structures were identified by mass spectrometry and (1) H NMR spectroscopy. Compared to a normal counter-current chromatography separation, the separation time and solvent consumption of elution-extrusion counter-current chromatography were reduced while the resolutions were still good. The established protocol is promising for the separation of natural products with great disparity of content in herbal medicines.

The chromatographic retention mechanism describing relationship between retention factor and concentration of Cu(2+) (l-phenylalanine)2 using chiral ligand mobile phase was investigated and eight mandelic acid derivatives were enantioseparated by chiral ligand exchange chromatography. The relationship between retention factor and concentration of the Cu(2+) (l-phenylalanine)2 complex was proven to be in conformity with chromatographic retention mechanism in which chiral discrimination occurred both in mobile and stationary phase. Different copper(II) salts, chiral ligands, organic modifier, pH of aqueous phase, and conventional temperature on retention behavior were optimized. Eight racemates were successfully enantioseparated on a common reversed-phase column with an optimized mobile phase composed of 6 mmol/L of l-phenylalanine or N,N-dimethyl-l-phenylalanine and 3 mmol/Lof copper(II) acetate or copper(II) sulfate aqueous solution and methanol.

Catalyzed by DDQ and NaNO2, the oxidative coupling between benzylic compounds and 1,3-dicarbonyls in the presence of molecular oxygen and HCOOH was developed. The 1% catalytic amount of DDQ is enough to complete the reaction. This system shows high efficiency and the coupling products are obtained in good to excellent yields within half an hour.

An efficient amidation has been developed of 1,3-diarylpropenes by carboxamides, sulfonamides, carbamates and anilines catalysed by 2,3-dichloro-5,6-dicyano-1,4-benzoquinone and benzoyl peroxide with molecular oxygen. The corresponding products were obtained in moderate to good yields.

Shenqi Jiangtang Granule, a well-known traditional Chinese herbal preparation, has been widely used for the treatment of type II diabetes mellitus. In this work, an ultrafiltration liquid chromatography with quadrupole time-of-flight mass spectrometry method was proposed for the rapid identification of bioactive ingredients from Shenqi Jiangtang Granule using α-glucosidase as an example. First, the chemical profile of this preparation was clarified, including 37 saponins, 17 flavonoids, 37 lignans, and seven other compounds. After incubation with α-glucosidase in vitro, the methanol extract with an IC50 value of 0.19 mg/mL exhibited significant inhibitory activity. Then, 18 specific binding peaks were screened, and 15 peaks were identified. Among these, ten compounds were reported to have potential α-glucosidase inhibitory activity for the first time. Subsequently, the inhibitory activities of these active compounds were evaluated by ultraviolet spectrophotometry with p-nitrophenyl α-d-glucopyranoside as a substrate. As a result, gomisin J and gomisin D exhibited stronger α-glucosidase inhibitory activities than other active compounds with IC50 values of 77.69 and 133.85 μM, respectively. The results demonstrated that the integrated ultrafiltration liquid chromatography with mass spectrometry method was an effective and powerful tool for the discovery of active ingredients in Shenqi Jiangtang Granule.

Four β-adrenergic blocking agents, including 1-[(1-methylethyl)amino]-3-phenoxy-2-propanol (1), 1-[(1-methylethyl)amino]-3-(3-methylphenoxy)-2-propanol (2), 1,1′-[1,4-phenylenebis(oxy)]bis[3-[(1-methylethyl)amino]-2-propanol (3) and 1,1′-[(4-methyl-1,2-phenylene)bis(oxy)]bis[3-[(1-methylethyl)amino]-2-propanol (4), were stereoselectively separated by countercurrent chromatography using di- n -hexyl l -tartrate and boric acid as chiral selector. The compounds (3) and (4) have four optical isomers since they contained two chiral centers. A two-phase solvent system composed of chloroform-0.05 mol L −1 of acetate buffer containing 0.10 mol L −1 of boric acid (1:1, v/v) was selected, in which 0.10 mol L −1 of di- n -hexyl l -tartrate was added in the organic phase as chiral selector. 20–42 mg of each racemate was stereoselectively separated by countercurrent chromatography in a single run with high purity of 96–98%, and the recovery of each separated compound reached around 87–93%. This is the first time report on successful stereoselective separation of optical isomeric compounds containing two chiral centers by countercurrent chromatography. At the same time, a chiral stationary phase was screened for analytical stereoselective separation of compounds (3) and (4) by high performance liquid chromatography.

The separation of ten epimeric aromatic acid (–)-menthol esters by countercurrent chromatography with hydroxypropyl-β-cyclodextrin as the mobile phase additive was investigated, and methods for the analysis of all the epimeric esters by reversed-phase high-performance liquid chromatography were established. A biphasic solvent system composed of n-hexane/20–70% methanol containing 50 mmol L−1 of hydroxypropyl-β-cyclodextrin (1:1, v/v) was selected, which provided high separation factors for five of the epimeric esters, and successful separations by countercurrent chromatography were achieved. The complete separation of five pairs of epimeric ester was obtained with the purity being over 98% for each peak fractions, as determined by high-performance liquid chromatography. The recovery of each analyte from the eluted fractions reached around 80–88%. This article is protected by copyright. All rights reserved

Preparative enantioseparations of α-cyclopentylmandelic acid and α-methylmandelic acid by high-speed countercurrent chromatography (HSCCC) and high performance liquid chromatography (HPLC) were compared using hydroxypropy-β-cyclodextrin (HP-β-CD) and sulfobutyl ether-β-cyclodextrin (SBE-β-CD) as the chiral mobile phase additives. In preparative HPLC the enantioseparation was achieved on the ODS C18 reverse phase column with the mobile phase composed of a mixture of acetonitrile and 0.10 mol L−1 phosphate buffer at pH 2.68 containing 20 mmol L−1 HP-β-CD for α-cyclopentylmandelic acid and 20 mmol L−1 SBE-β-CD for α-methylmandelic acid. The maximum sample size for α-cyclopentylmandelic acid and α-methylmandelic acid was only about 10 mg and 5 mg, respectively. In preparative HSCCC the enantioseparations of these two racemates were performed with the two-phase solvent system composed of n-hexane-methyl tert.-butyl ether-0.1 molL−1 phosphate buffer solution at pH 2.67 containing 0.1 mol L−1 HP-β-CD for α-cyclopentylmandelic acid (8.5:1.5:10, v/v/v) and 0.1 mol L−1 SBE-β-CD for α-methylmandelic acid (3:7:10, v/v/v). Under the optimum separation conditions, total 250 mg of racemic α-cyclopentylmandelic acid could be completely enantioseparated by HSCCC with HP-β-CD as a chiral mobile phase additive in a single run, yielding 105-110 mg of enantiomers with 95-98% purity and 85-90% recovery. But, no complete enantioseparation of α-methylmandelic acid was achieved by preparative HSCCC with either of the chiral selectors due to their limited enantioselectivity. In this paper preparative enantioseparation by HSCCC and HPLC was compared from various aspects.

This work concentrates on the separation mechanism and application of chiral ligand exchange high-speed countercurrent chromatography (HSCCC) in enantioseparations, and comparison with traditional chiral ligand exchange high performance liquid chromatography (HPLC). The enantioseparation of ten aromatic α-hydroxyl acids were performed by these two chromatographic methods. Results showed that five of the racemates were successfully enantioseparated by HSCCC while only three of the racemates could be enantioseparated by HPLC using a suitable chiral ligand mobile phase additive. For HSCCC, the two-phase solvent system was composed of butanol-water (1:1, v/v), to which N-n-dodecyl-L-proline was added in the organic phase as chiral ligand and cupric acetate was added in the aqueous phase as a transition metal ion. Various operation parameters in HSCCC were optimized by enantioselective liquid-liquid extraction. Based on the results of the present studies the separation mechanism for HSCCC was proposed. For HPLC, the optimized mobile phase composed of aqueous solution containing 6 mmol L−1 L-phenylalanine and 3 mmol L−1 cupric sulfate and methanol was used for enantioseparation. Among three ligands tested on a conventional reverse stationary phase column, only one was found to be effective. In the present studies HSCCC presented unique advantages due to its high versatility of two-phase solvent systems and it could be used as an alternative method for enantioseparations.

The influence of chlorine substituents in chiral separation of three racemic 2-(chlorophenyl)propanoic acids by countercurrent chromatography using hydroxypropyl-β-cyclodextrin as a chiral additive were mainly investigated in the present paper, including 2-(2-chlorophenyl)propanoic acids, 2-(3-chlorophenyl)propanoic acids and 2-(4-chlorophenyl)propanoic acids. The influences of chromatographic conditions on the retention behavior were studied by enantioselective liquid-liquid extraction experiments using the methodology of response surface It was found that 2-(3-chlorophenyl)propanoic acids could be successfully chiral separated by countercurrent chromatography, while no resolution was achieved for racemic 2-(2-chlorophenyl)propanoic acids and 2-(4-chlorophenyl)propanoic acids under optimized separation conditions. The formation of 1:1 stoichiometric inclusion compounds between 2-(3-chlorophenyl)propanoic acids and HP-β-CD was determined by UV spectra measurements. The inclusion constants for 2-(3-chlorophenyl)propanoic acids and HP-β-CD were determined by the Benesi-Hildebrand equation. Meanwhile, the inclusion constants of 2-(3-chlorophenyl)-propanoic acid enantiomer and HP-β-CD were obtained by the pesudophase retention equation in countercurrent chromatography. Furthermore, the inclusion interactions of the three racemates with HP-β-CD were also investigated by the molecular docking. The results obtained from UV spectra measurements and molecular docking showed that the racemate with chlorine substituents in meta-position presented the highest enantiorecognition while the racemates with chlorine substituents in ortho-position had the lowest enantiorecognition. The above results further indicated that forming a stable inclusion complex between racemate and chiral selector is a prerequisite for a successful enantioseparation and at the same time, the difference in inclusion capacity between the two enantiomers is also essential for the enantioseparation.

This study established a rapid and reliable approach using liquid chromatography-tandem mass spectrometry for the simultaneous determination of cinnamic acid, vanillic acid and protocatechuic acid in rat plasma. This is the first report on a comparative pharmacokinetic study of dispensing granules and standard decoction of Cinnamomum cassia twigs in rats. After liquid-liquid extraction by ethyl acetate, the plasma samples were subjected to LC-MS/MS for multiple reaction monitoring. The standard curves showed good linear regression (r2 > 0.9991) in the range of 10.0-16000 ng/mL. The intra- and inter-day accuracy and precision were found to be within 15% of the nominal concentration. The recoveries of the three phenolics ranged from 88.7 to 105.7%. Finally, this approach was successfully applied to pharmacokinetic analysis of the three phenolics after oral administration of standard decoction and dispensing granules of C. cassia twigs in rats. Although the values of AUC0-t of vanillic acid and protocatechuic acid in standard decoction group were larger than those of the dispensing granule group, no significant difference was observed for the two groups. Of note, the elimination rates of vanillic acid were slower in the standard decoction group than the dispensing granule group.