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The current study highlights the advantages of using the quality by design (QbD) approach to synthesise and optimize SERS substrates for the detection of Hg2+.

Chromatographic fingerprinting provides effective technical means for quality evaluation of traditional Chinese medicine.

Caesalpinia sappan L. (C. sappan L.), a traditional Chinese medicine, has been widely used to treat bruises and dysmenorrhea, performing pharmacological activities such as anti-inflammatory and anti-tumor. C. sappan L. has been reported to contain compounds such as protosappanins, brazilins, and homoisoflavones. In the pre-experiments, we discovered that there were many unstable components in the characteristic chromatogram of C. sappan L. Here, silica-gel column chromatography, high-speed counter-current chromatography, and preparative high-performance liquid chromatography were combined and applied to isolate the unstable components from alcohol extract of C. sappan L. The results showed that four unstable compounds were collected with the purity higher than 95.0%, characterized as episapponal, brazilin, sapponal, and 4-O-methylsapponal by hydrogen-1 and carbon-13 nuclear magnetic resonance. Based on the above results, the characteristic chromatogram of C. sappan L. was established, and the characteristic peaks were identified. These results provided a theoretical basis for the quality assessment of C. sappan L.

An intelligently and globally extracting feature information method, different from a multi-wavelength fusion method, is used in HPLC-DAD chromatography. It combined with chemometrics is suitable for quality control of traditional Chinese medicines.

Objective: To study the blood protective effect of platelet autotransfusion in cardiopulmonary bypass. Methods: In this study, 80 patients with cardiopulmonary bypass heart surgery in our hospital from January 2018 to July 2019 were divided into two groups. The control group was not given platelet separation, the study group was given platelet separation after anesthesia induction, platelet rich plasma was extracted before heparinization for cardiopulmonary bypass and heparin was recycled. The blood clotting function, postoperative recovery, blood transfusion volume and postoperative drainage volume were analyzed before operation, 1H, 24h and 48h after operation. Result: Compared with the control group, the volume of blood flow, blood plasma and platelet transfusion in the study group decreased significantly in the first hour after operation, the difference was statistically significant (P < 0.05). The recovery of the study group was slightly better than that of the control group, and the difference was not statistically significant (P > 0.05). Conclusion: Autotransfusion of platelets can alleviate the coagulation function of patients undergoing cardiopulmonary bypass, reduce the amount of postoperative bleeding and allogeneic blood transfusion, and protect the blood.

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 method for quantitative determination of the main active ingredient in traditional Chinese medicine was developed and validated, using liquid-liquid chromatography combined with high performance ...

A rapid, sensitive, environmental friendly dual preconcentration method by combining micro matrix solid-phase dispersion extraction with field-enhanced sample injection and micelle to cyclodextrin stacking has been developed for the determination of furocoumarins. Molecular sieve, KIT-6, was used as an adsorbent in micro matrix solid-phase dispersion process. The important parameters affecting off-line and on-line CE preconcentration efficiency were optimized. Under the optimal experimental conditions, all analytes showed good linearity (R2 > 0.999). The LODs of notopterol, isoimperatorin and imperatorin were 0.1 μg/mL, 1.2 mg/kg and 1.0 mg/kg respectively. Compared with the normal CE method, the enrichment times were up to 300. Moreover, Angelicae Dahuricae Radix was used as the mode of complex solid sample matrix to demonstrate the prospect of application of this methodology. The results showed the proposed strategy is promising for determining trace furocoumarins in complex matrix samples, which might be applied as a powerful and economic tool in monitoring illegal cosmetic adding. This article is protected by copyright. All rights reserved.

In traditional Chinese therapy, the decoction of herbal products is commonly used as a therapeutic drug, which takes water as the extraction medium and contains many hydrophilic compounds. The main...

A rhodium-catalyzed tandem intermolecular hydroacylation of terminal alkyne-substituted secondary alcohols with chelating aldehydes and deconjugative isomerization of the resulting α,β-unsaturated ketones for the preparation of synthetically valuable 1,4-diketones is developed. Of note, the completely atom-economical protocol can be extended to the alkynyl alcohols with appropriate carbon chain length, from which 1,5-diketone and 1,6-diketone were obtained involving the short-range deconjugative isomerization. Given its robustness, broad substrate scope, and excellent regioselectivity, this approach provides a promising platform for the synthesis of diverse diketones.

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.

As a liquid-liquid partition chromatography, counter-current chromatography has advantages in large sample loading capacity without irreversible adsorption, which has been widely applied in separation and purification fields. The main factors, including partition coefficient, two-phase solvent systems, apparatus, and operating parameters greatly affect the separation process of counter-current chromatography. To promote the applications of counter-current chromatography, it is essential to develop theoretical research to master the principles of counter-current chromatographic separations so as to achieve predictions before laborious trials. In this article, recent progress about separation prediction methods are reviewed from a point of the steady and unsteady state of the mass transfer process of counter-current chromatography and its mass transfer characteristics, and then it is divided into three aspects: prediction of partition coefficient, modeling the thermodynamic process of counter-current chromatography, and modeling the dynamic process of counter-current chromatography.

Two new nor-lignans, pulvin A (1) and moellenoside C (2), along with two known compounds (3–4) were isolated from the whole plant of Selaginella pulvinate (Hook. & Grev.) Maxim. The structures of t...

A simple, efficient, and sensitive strategy by coupling matrix solid-phase dispersion with ultra high performance liquid chromatography quadrupole time-of-flight mass spectrometry was proposed to extract and determine three types of components (including seven analytes) in Chinese patent medicines Chenxiangqu. The highly ordered mesoporous material Fe-SBA-15 synthesized under weakly acidic conditions was selected as a dispersant in matrix solid phase dispersion extraction for the first time. Several parameters including the mass ratio of sample to dispersant, the type of dispersant, the grinding time, and the elution condition were investigated in this work. Under the optimized conditions, 20 compounds were identified by quadrupole time-of-flight mass spectrometry and seven analytes were quantified. The results demonstrated that the developed method has good linearity (r > 0.9995), and the limits of detection of the analytes were as low as 0.55 ng/mL. The recoveries of all seven analytes ranged from 97.6 to 104.6% (relative standard deviation < 3.4%). Finally, the improved method was successfully applied to determination of five batches of Chenxiangqu samples, which provided a robust method in quality control of Chinese patent medicines Chenxiangqu. The developed strategy also shows its great potential in analysis of complex matrix samples.

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.

Mediated by 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), a novel oxidative coupling of hydrazones and 1,3-diarylpropenes has been disclosed to generate appealing β,γ-unsaturated hydrazones, which further undergo 5-exo-trig or 6-endo-trig cascade cyclization to give the respective 1,2-dihydropyridazines or pyrazoles selectively under metal-free conditions. The mechanisms of the coupling and subsequent cyclization are proposed.

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.

Despite the recent increase in the development of bioactive molecules in the drug industry, the enormous chemical space and lack of productivity are still important issues. Additional alternative approaches to screen and locate bioactive molecules are urgently needed. Label-free bio-affinity mass spectrometry (BA-MS) provides opportunities for the discovery and development of innovative drugs. This review provides a comprehensive portrayal of BA-MS techniques and of their applications in screening and locating bioactive molecules. After introducing the basic principles, alongside some application notes, the current state-of-the-art of BA-MS-assisted drug discovery is discussed, including native MS, size-exclusion chromatography-MS, ultrafiltration-MS, solid-phase micro-extraction-MS, and cell membrane chromatography-MS. Finally, several challenges and limitations of the current methods are summarized, with a view to potential future directions for BA-MS-assisted drug discovery. © 2019 Wiley Periodicals, Inc. Mass Spec Rev.

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.

Type 2 diabetes is a chronic metabolic disorder affecting millions of people worldwide, and new drug leads are urgently needed. The dried stem of Dendrobium officinale is used in China to alleviate the symptoms of diabetes, but investigations of its bioactive components have been limited. We have applied a simple, fast, and effective method, combining triple high-resolution profiling of radical scavenging and α-glucosidase and α-amylase inhibition with high-performance liquid chromatography - photodiode-array detection - high-resolution mass spectrometry - solid-phase extraction - nuclear magnetic resonance spectroscopy (HR-bioassay/HPLC-PDA-HRMS-SPE-NMR). Two new compounds, 3,4,4′-trihydroxy-5-methoxybibenzyl (5) and 3,4-dihydroxy-3′,4′,5-trimethoxybibenzyl (8a), along with fourteen known compounds, were identified. Twelve of these were associated with radical scavenging activity, six with α-glucosidase inhibition, and one with α-amylase inhibition. Compound 5, dendrocandin U, and 3,4-dihydroxy-4′,5-dimethoxybibenzyl (12) are reported as α-glucosidase inhibitors for the first time, and 12 is furthermore reported as an α-amylase inhibitor for the first time.

Background: Aflatoxins (AFs) have attracted increasing amounts of attention in recent years for their high toxicity. Previously, AF-polluted Chinese Patent Medicines (CPMs) were ignored, but their quality, safety, and efficacy might be influenced by the toxic compounds. Objective: The objective is to develop a simple, low-cost, and efficient method for quantitative analysis of aflatoxins B1 (AFB1) and B2 (AFB2) in CPMs to ensure their safe use. Methods: A simple, modified, magnetic nanoparticle-based solid-phase extraction (SPE) combined with HPLC-fluorescence detection for the determination of trace amounts of AFB1 and AFB2 was established. The main parameters affecting the efficiency of modified magnetic nanoparticle-based SPE, such as pH of sample solution, adsorbent amount, adsorption time, and desorption condition, were investigated. Results: Under the optimum conditions, AFB1 and AFB2 were linear in the ranges of 0.3–10 and 0.04–3 ng/mL with the correlation coefficient (R) of 0.9998 and 0.9999, respectively. Their intraday precisions were 1.16 and 2.30% and the interday precisions were 1.28 and 1.87% for AFB1 and AFB2, respectively. The developed SPE was applied for AFB1 and AFB2 extraction in three commercially available Fupuganmao granule samples, and the results were compared with the official immunoaffinity column chromatography method. Conclusions: The method provided a preferable candidate for the determination approach of AF measurement in CPMs. Highlights: Amine-functionalized magnetic nanoparticles were successfully applied to SPE for adsorbing AFB1 and AFB2 in CPMs for the first time.

Ethnopharmacological relevance Achyranthes bidentata Blume (AB) is a well-known traditional Chinese medicine for treating osteoporosis and bone fracture. In the current, researches on pharmacological mechanism of AB mostly focused on molecular pathways, knowledge about its metabolic signatures is largely unclear. This study aims to develop an integrative metabolomics and metallomic approach for deciphering the biochemical basis of anti-osteoporosis effects of raw and salt-processed AB. Method Gas chromatography–mass spectrometry (GC–MS) and inductively coupled plasma mass spectrometry (ICP-MS) were combined for metabolomic and metallomic profiling of rats serum, liver and kidney derived from the sham group, model group, E2, raw and salt-processed AB treated groups. Meanwhile, micro-CT and biomechanical analysis were carried out to ensure the success of the osteoporosis model and to validate the anti-osteoporosis effect of raw and salt-processed AB. Partial least squares discriminant analysis (PLS-DA) was employed to screen potential biomarkers and the MetaboAnalyst and KEGG PATHWAY Database were used to investigate the metabolic pathway. Results Raw and salt-processed AB protected the rats against osteoporosis, as evidenced by the restoration of the alkaline phosphatase activity, osteocalcin concentration, urine calcium/creatinine ratio and urine phosphorus/creatinine ratio. The combination of PCA and PLS-DA revealed deviations in ninety-four differential biomarkers between raw AB treated group and model group. The identified biomarkers were primarily engaged in the metabolic pathways including galactose metabolism, urea cycle, arginine and proline metabolism, alanine metabolism, lactose degradation, ammonia recycling and glycine and serine metabolism. The levels of these biomarkers showed significant alterations and a tendency to be restored to normal values in raw and salt-processed AB treated osteoporosis rats. Of note, the levels of trace elements, such as Zn, Se, Mn, Cu and Fe, were elevated after raw and salt-processed AB treatment. Finally, a correlation network diagram was constructed to show the biomarkers perturbed by raw and salt-processed AB. Conclusion Our findings indicate that raw and salt-processed AB has positive effects on osteoporosis rats. Meanwhile, metabolomic and metallomic method coupled with metabolites enrichment analysis and pattern recognition serves as a useful tool for revealing the action mechanism of traditional Chinese medicine.

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).

A novel analytical quality by design approach for developing a chromatographic fingerprint was established for analyzing complex traditional Chinese medicine, using a licorice standard decoction as an example. Considering the characteristics of integrity and ambiguity, the resolution of eight common peaks, total peak number, capacity factor distributions, and peak purity were selected as potential critical method attributes for assessing the quality of the chromatographic fingerprint. A central composite design was used to evaluate the relationship between critical method attributes and critical method parameters, including column temperature, wavelength, flow rate, formic-acid concentration, and gradient parameters. A standard probability method was employed to calculate the design space of the fingerprint analysis parameters and evaluate the robustness of the methodology. The optimized high-performance liquid chromatography fingerprint conditions were acetonitrile and 0.1% formic acid water gradient elution (0-5 min, 5-19% A; 5-10 min, 19% A; 10-50 min, 19-42% A; 50-54 min, 42-100% A; 54-60 min, 100% A), column temperature 25±5°C, detection wavelength 265 nm. The design space of fingerprint analytical method based on the analytical quality by design approach not only met the requirements of the fingerprint analysis, but also improved the robustness and applicability of the fingerprint method.

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.

OBJECTIVE: To explore genetic mutation types and their correlation with clinical phenotypes in Uighur patients with aortic disease in Kashgar (Xinjiang Uighur Autonomous Region, China). METHODS: We examined 37 pathogenic genes in 19 Uighur families with aortic diseases including Marfan syndrome from Kashgar using next generation sequencing, and the results were confirmed by Sanger sequence in the first relatives. RESULTS: This study included 19 families with aortic diseases, in whom a total of 23 variants were identified, and 11 (57.89%) probands had one or more variants. Among them, definite pathogenic mutation was detected in one patient (5.26%), variants of uncertain significance (VUS) were found in 8 (42.11%), and benign/likely benign variants were detected in 7 (36.84%). The 23 variants identified included one (5.26%) pathogenic variant, 14 (60.87%) VUS, and 8 (34.78%) benign/likely benign variants. The 14 VUS were analyzed by prediction with SIFT and Polyphen2 HDIV, which identified 6 (42.86%) variants as deleterious/possibly damaging; all the 8 benign/likely benign variants were predicted to be deleterious/possibly damaging. CONCLUSIONS: We detected 23 genetic variants in the 19 Uighur families with aortic diseases, and 22 of these variants remain to be verified by more patient data in future studies.

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.

A sensitive capillary electrophoresis strategy was developed by combining salting-out assisted liquid-liquid extraction and online three-step stacking to detect trace quantities of antitumour indole alkaloids in complex biological samples. The proposed strategy fully exploits these two technologies such that extraction, online stacking and separation are combined in a fast and efficient manner. First, salting-out assisted liquid-liquid extraction was used to extract three indole alkaloids (vinblastine sulfate, catharanthine sulfate and vindoline) from complex biomasses. An appropriate volume of acetonitrile (ACN) was mixed with a faecal aqueous solution to precipitate proteins. The mixed solution was vortexed, followed by the addition of ammonium sulfate ((NH

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.

A simple, convenient, and sensitive method that involves combining matrix solid-phase dispersion extraction with field-amplified sample stacking in capillary electrophoresis has been developed for the determination of organic acids in a complex solid matrix. Mesoporous molecular sieve, MCM-48, was synthesized by a hydrothermal method and selected as the adsorbent in matrix solid-phase dispersion. After fast extraction, the enriched analytes were back-extracted into a basic aqueous solution for field-amplified sample stacking in capillary electrophoresis. Parameters that affect extraction efficiency and sample stacking were optimized. Under the optimal conditions, approximately 42-, 49-, and 56-fold sensitivity enhancements were achieved for danshensu, protocatechuic acid, and cinnamic acid, respectively, when compared to normal injection. A satisfactory correlation coefficient (r > 0.99) was obtained. Both intra- and interday precision were lower than 2.53%. And the limits of detection of the three organic acids ranged between 0.01 and 0.029 μg/mL. Finally, the newly proposed method was successfully applied to the determination of organic acids in Fufang Danshen tablets, which indicates its great potential in analyzing organic acids in a complex matrix.

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.

Recycling countercurrent chromatography was successfully applied to the resolution of 2-(4-bromomethylphenyl)propionic acid, a key synthetic intermediate for synthesis of nonsteroidal anti-inflammatory drug loxoprofen, using hydroxypropyl-β-cyclodextrin as chiral selector. The two-phase solvent system composed of n-hexane/n-butyl acetate/0.1 mol/L citrate buffer solution with pH 2.4 (8:2:10, v/v/v) was selected. Influence factors for the enantioseparation were optimized, including type of substituted β-cyclodextrin, concentration of hydroxypropyl-β-cyclodextrin, separation temperature, and pH of aqueous phase. Under optimized separation conditions, 50 mg of 2-(4-bromomethylphenyl)propionic acid was enantioseparated using preparative recycling countercurrent chromatography. Technical details for recycling elution mode were discussed. The purities of both the S and R enantiomers were over 99.0% as determined by high-performance liquid chromatography. The enantiomeric excess of the S and R enantiomers reached 98.0%. The recovery of the enantiomers from eluted fractions was 40.8-65.6%, yielding 16.4 mg of the S enantiomer and 10.2 mg of the R enantiomer. At the same time, we attempted to enantioseparate the anti-inflammatory drug loxoprofen by countercurrent chromatography and high-performance liquid chromatography using a chiral mobile phase additive. However, no successful enantioseparation was achieved so far.

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.