In vitro anti-platelet aggregation effects of fourteen fruits and vegetables.

In the present study, the anti-platelet aggregation activity of 14 vegetables and fruits was tested in vitro. The aqueous, 90% ethanol and ethyl acetate extracts, as well as concentrated juices of 14 foods (fruits and vegetables) were prepared, and the anti-platelet aggregation activity of those extracts was analyzed on a platelet aggregation analyzer in vitro with adenosine 5'-diphosphate (ADP), bovine thrombin (THR) and arachidonic acid (AA) as aggregation inducers, respectively. Aspirin (ASP) was used as the positive control. A number of the tested foods had inhibitory effects in concentration-dependent manner on platelet aggregations induced by various agonists. Especially, some foods such as lemon, leek, garlic, scallion, ginger, tomato and grapefruit showed good anti-platelet aggregation effect similar or higher than that of positive control group i.e. aspirin (ASP). The results of present study provide scientific reference for reasonable selection of daily dietary with supplementary curative effects or prevention of cardiovascular diseases (CVD).

Adsorbed hollow fiber-based biological fingerprinting for the discovery of platelet aggregation inhibitors from Danshen-Honghua decoction.

For lead compound discovery from natural products, hollow fiber cell fishing with chromatographic analysis is a newly developed method. In this study, an adsorbed hollow fiber-based biological fingerprinting method was firstly developed to discover potential platelet aggregation inhibitors from Danshen-Honghua decoction. Platelets were seeded on the fiber and their survival rate was tested. Results indicated that more than 92% platelets survived during the whole operation process. Ranitidine and tirofiban were used as positive and negative control respectively to verify the reliability of the presented approach. The main variables such as amount of extract and stirring time that affect the adsorbed hollow fiber-based biological fingerprinting process were optimized, and the repeatability of this method was also investigated. Finally, 12 potential active compounds in Danshen-Honghua decoction were successfully detected using the established approach and structures for nine of them were tentatively identified by liquid chromatography with mass spectrometry analysis. In addition, the in vitro platelet aggregation inhibition test was carried out for five of the nine hit compounds, and three active components, namely, lithospermic acid, salvianolic acid A, and salvianolic acid B were confirmed. These results proved that the proposed method could be an effective approach for screening platelet inhibitors from plant extracts.

Comparing coagulation activity of Selaginella tamariscina before and after stir-frying process and determining the possible active constituents based on compositional variation.

CONTEXT: Selaginella tamariscina (P. Beauv.) Spring (Selaginellaceae) (ST) has been widely used in China as a medicine for improving blood circulation. However, its processed product, S. tamariscina carbonisatus (STC), possesses opposite haemostatic activity. OBJECTIVE: To comprehensively evaluate the activity of ST and STC on physiological coagulation system of rats, and seek potential active substances accounting for the activity transformation of ST during processing. MATERIALS AND METHODS: The 75% methanol extracts of the whole grass (fine powder) of ST and STC were prepared, respectively. Male Sprague-Dawley rats were randomly divided into five groups: control group, model group, model + ST group, model + STC group and positive control group (model + Yunnanbaiyao). The duration of intragastric administration was 72 h at 12 h intervals. Haemorheology parameters were measured using an LB-2 A cone-plate viscometer and the existed classic methods, respectively. SC40 semi-automatic coagulation analyzer was employed to determine coagulation indices. Meanwhile, HPLC and LC-MS were applied for chemical analyses of ST and STC extracts. RESULTS: STC shortened tail-bleeding time, increased whole blood viscosity (WBV) and plasma viscosity (PV), decreased erythrocyte sedimentation rate blood (ESR), reduced activated partial thromboplastin time (APTT) and increased the fibrinogen (FIB) content in the plasma of bleeding model rats. Although ST could shorten APTT and TT, the FIB content was significantly decreased by ST. Dihydrocaffeic acid with increased content in STC vs. ST showed haemostatic activity for promoting the platelet aggregation induced by collagen and trap-6, and reducing APTT and PT significantly with a concentration of 171.7 µM in vitro. Amentoflavone with reduced content in STC vs. ST inhibited ADP and AA-induced platelet aggregation significantly with a concentration of 40.7 µM. DISCUSSION AND CONCLUSIONS: As the processed product of ST, STC showed strong haemostatic activity on bleeding rat through regulating the parameters involved in haemorheology and plasma coagulation system. Two active compounds, dihydrocaffeic acid and amentoflavone, might be partially responsible for the haemostatic and anticoagulant activity of STC and ST, respectively.

SDS-PAGE and 2-DE protein profiles of Ganoderma lucidum from different origins.

Ganoderma lucidum (Chizhi in Chinese) is one of the most valuable and widely used medicinal fungi in traditional Chinese medicines (TCMs). Most of previous studies were focused on the triterpenoids and polysaccharides of G. lucidum, whereas less attention had been paid on the protein, which is another bioactive compound in it. In the present study, protein maps of fourteen G. lucidum samples were comprehensively analyzed by sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE). The results indicated that there were significant differences in protein profiles of G. lucidum samples from different origins. Furthermore, previous reported bioactive proteins from the fruiting bodies of G. lucidum, were mainly distributed in 4 taxa (A, B, C and D) based on their molecular weights on the 2-DE maps. The proteins should be considered as marker for the quality control of G. lucidum, because the proteomic variation may affect on their pharmacological activities.

Evaluation of interactions between RAW264.7 macrophages and small molecules by capillary electrophoresis.

In this study, the affinity interactions between RAW 264.7 macrophages and three small molecules including naringin, oleuropein and paeoniflorin were evaluated by affinity capillary electrophoresis (ACE), partial filling affinity capillary electrophoresis (PFACE) and frontal analysis capillary electrophoresis (FACE), respectively. The result indicated that ACE (varying concentrations of cell suspension were filled in the capillary as receptor) may not be suitable for the evaluation of interactions between cell and small molecules due to the high viscosity of cell suspension; PFACE can qualitatively evaluate the interaction, but the difference in viscosity between RAW264.7 suspension and buffer effects on the liner relationship between filling length and injection time, which makes the calculation of binding constant difficult. Furthermore, based on the PFACE results, naringin showed stronger interaction with macrophages than the other two molecules; taking advantage of the aggregation phenomenon of cell induced by electric field, FACE was successfully used to determine the stoichiometry (n = 5×109 ) and binding constant (Kb = 1×104 L/mol) of the interaction between RAW264.7 and naringin.

Recent applications of hydrophilic interaction liquid chromatography in pharmaceutical analysis.

Hydrophilic interaction liquid chromatography, an alternative liquid chromatography mode, is of particular interest in separating hydrophilic and polar ionic compounds. Compared with traditional liquid chromatography techniques, hydrophilic interaction liquid chromatography offers specific advantages mainly including: (1) relatively green and water-soluble mobile phase composition, which enhances the solubility of hydrophilic and polar ionic compounds; (2) no need for ion-pairing reagents and high content of organic solvent, which benefits mass spectrometry detection; (3) high orthogonality to reverse-phase liquid chromatography, well adapted to two-dimensional liquid chromatography for complicated samples. Therefore, hydrophilic interaction liquid chromatography has been rapidly developed in many areas over the past decades. This review summarizes the recent progress (from 2012 to July 2016) of hydrophilic interaction liquid chromatography in pharmaceutical analysis, with the focus on detecting chemical drugs in various matrices, charactering active compounds of natural products and assessing biotherapeutics through typical structure unit. Moreover, the retention mechanism and behavior of analytes in hydrophilic interaction liquid chromatography as well as some novel hydrophilic interaction liquid chromatography columns used for pharmaceutical analysis are also described.

Molecularly imprinted polymers for the selective extraction of tiliroside from the flowers of Edgeworthia gardneri (wall.) Meisn.

Nano-sized molecularly imprinted polymers for tiliroside were successfully prepared by a precipitation polymerization method. Acrylamide, ethylene glycol dimethacrylate, azobisisobutyronitrile, and acetonitrile/dimethyl sulfoxide were used as functional monomer, cross-linker, initiator, and porogen, respectively. The structural features and morphological characterization of tiliroside-imprinted polymers were characterized by Fourier transform infrared spectroscopy and scanning electron microscopy, respectively. The adsorption experiments indicated that the tiliroside-imprinted polymers exhibited high selective recognition property to tiliroside. Scatchard analysis indicated that the homogeneous-binding sites were formed in the polymers. The selectivity test revealed that the adsorption capacity and selectivity of polymers to tiliroside was significantly higher than that of rutin, astragalin, and kaempferol. Finally, the tiliroside-imprinted polymers were employed as adsorbents in solid-phase extraction for the extraction of tiliroside from the ethyl acetate extract of the flowers of Edgeworthia gardneri (wall.) Meisn. The results demonstrated that the extraction recoveries of tiliroside ranged from 69.3 to 73.5% by using tiliroside-imprinted polymers coupled with solid-phase extraction method. These results indicated that the tiliroside-based molecularly imprinted solid-phase extraction method was proven to be an effective technique for the separation and enrichment of tiliroside from natural medicines.

[Applications of immobilization techniques in the screening of active constituents from traditional Chinese medicines].

It has been an active approach to screen the active ingredients in traditional Chinese medicines (TCMs) according to the affinity property between small molecule compounds and biomaterials such as cells, bacteria and proteins. On the other hand, the biomaterials can be immobilized on a solid support before the screening procedure. The immobilization method not only can maintain the biological activities of biomaterials, but also have other advantages such as high efficiency, simple operation, easy to be continuous and automatic, etc. Carrier materials (solid supports) for the immobilization including silica gel, magnetic materials, hollow fiber, and the surface plasma resonance sensor chips have been used to immobilize biomaterials and successfully applied in the screening of active ingredients from TCMs. In this paper, applications of immobilization techniques in the screening of active components from TCMs were reviewed to provide a scientific reference to the future applications.

Differential proteomic analysis of platelets suggested target-related proteins in rabbit platelets treated with Rhizoma Corydalis.

CONTEXT: Corydalis yanhusuo W.T. Wang (Papaveraceae) (Rhizoma Corydalis) showed inhibitory effects on rabbit platelet aggregation induced by ADP, thrombin (THR) or arachidonic acid (AA). OBJECTIVE: This study separates and identifies the possible target-related platelet proteins and suggests possible signal cascades of RC antiplatelet aggregation. MATERIALS AND METHODS: Based on comparative proteomics, the differentially expressed platelet proteins treated before and after with 50 mg/mL RC 90% ethanol extract (for 15 min at 37 °C) were analyzed and identified by two dimensional gel electrophoresis (2-DE) and MALDI-TOF-MS/MS. To further verify the possible signalling pathways of RC antiplatelet aggregation function, the concentration of calcium (Ca2+) was measured by Fura-2/AM fluorescence (Ex 340/380 nm, Em 500 nm) (RC final concentrations of 0.0156-0.1563 mg/mL), the levels of P-selectin and cyclic guanosine monophosphate (cGMP) were quantified by ELISA (OD. 450 nm) (RC final concentrations of 0.0156-1.5625 mg/mL), and the 5-hydroxytryptamine (5-HT) level was measured using ortho-phthalaldehyde (OPT) fluorescence (Ex 340 nm, Em 470 nm) (RC final concentrations of 0.3125-1.5625 mg/mL). RESULTS: The expression of 52 proteins were altered in rabbit platelets after the treatment and the MALDI-TOF-MS analysis indicated that those proteins include 12 cytoskeleton proteins, 7 cell signalling proteins, 3 molecular chaperone proteins, 6 proteins related to platelet function, 16 enzymes and 7 other related proteins. Furthermore, RC extract could decrease the levels of 5-HT [inhibition rate of 96.80% (p < 0.05, vs. THR-activated group) treated with 0.7813 mg/mL of RC], Ca2+ [172.73 ± 5.07 to 113.56 ± 5.46 nM (p < 0.001, vs. THR-activated group) treated with 0.0313 mg/mL of RC] and P-selectin [13.48 ± 0.96 ng/3 × 108 to 11.64 ± 0.17 ng/3 × 108 (p < 0.05, vs. THR-activated group) treated with 0.0156 mg/mL of RC], and increase in cGMP level [38.93 ± 0.57 to 50.26 ± 4.05 ng/3 × 108 (p < 0.05, vs. THR-activated group) treated with 1.5165 mg/mL of RC] in ADP (10 µmol/L), THR (0.25 u/mL) or AA-(0.205 mmol/L) activated rabbit platelets. DISCUSSION AND CONCLUSION: The present study indicated that P2Y12 receptor might be one of the direct target proteins of RC in platelets. The signal cascades network of RC after binding with P2Y12 receptor is mediating Gαi proteins to activate downstream signalling pathways (AC and/or PI3K signalling pathways) for the inhibition of platelet aggregation.

Evaluation of affinity interaction between small molecules and platelets by open tubular affinity capillary electrochromatography.

In this paper, an open tubular affinity capillary electrochromatography (OT-ACEC) was developed by physical adsorption of rabbit platelets on the inner surface of capillary. The interactions between small molecules include adenosine diphosphate (ADP) (positive control), protocatechuic acid (negative control) and seven natural products (salvianolic acid B, salvianic acid A sodium, hydroxysafflor yellow A, ferulic acid, chlorogenic acid, sinapic acid, caffeic acid) and platelets were evaluated by their retention factors and binding constants obtained based on peak-shift assay. Then, the activities of anti-platelet aggregation induced by thrombin (THR), ADP and arachidonic acid (AA) for those small molecules (except ADP) were evaluated by turbidimetric method. The results indicate that: (i) ADP, a platelet aggregation inducer, had strong interaction with platelet, while protocatechuic acid that had no inhibition on platelet aggregation behaved no specific interaction; (ii) there was a positive correlation between the anti-platelet aggregation activities of small molecules and their interactions with platelet, generally those compounds with higher binding constants with platelet exhibited higher activities. Therefore, the OT-ACEC method developed in the present study can be a potential method to evaluate affinity interactions between small molecules and platelets, so as to predict the biological activities such as anti-platelet aggregation for the small molecules.

Simultaneous screening and analysis of antiplatelet aggregation active alkaloids from Rhizoma Corydalis.

CONTEXT: The rising problem of atherosclerosis and ischemic heart disease emphasizes the need to look for new antithrombotic components with effective modes of action. Corydalis yanhusuo (Y.H. Chou & Chun C. Hsu) W.T. Wang ex Z.Y. Su & C.Y. Wu (Papaveraceae) (Rhizoma Corydalis) has been used in the traditional medicines for the treatment of cardiovascular disease. OBJECTIVE: The antiplatelet aggregation compounds in Rhizoma Corydalis were screened to validate its traditional medicinal use. MATERIAL AND METHODS: Total alkaloid extract (TAE) of Rhizoma Corydalis was obtained by refluxing 100 g Rhizoma Corydalis powder with 600 mL 70% ethanol, and purified by acidification (20% HCl) and alkalization (5 M NaOH) process. Potential antiplatelet aggregation compounds in TAE were screened by a method involving platelet bio-specific extraction and HPLC-DAD/LC-MS analysis. Further in vitro antiplatelet aggregation activity confirmation of TAE and seven main alkaloids were achieved by turbidimetry method within 3 h after blood collection from rabbit carotid artery, and all the test drugs were at the concentration range of 25-350 µg/mL. Finally, HPLC-DAD was employed for the quantitative determination of seven main components in TAE. RESULTS: Five alkaloids, identified as glaucine, dehydrocorydaline, canadine, tetrahydrocoptisine and corydaline, can be specifically extracted with platelets. The results indicated that all these five alkaloids can inhibit thrombin-induced platelet aggregation in a low dose (IC50 of glaucine, dehydrocorydaline, canadine, tetrahydrocoptisine and corydaline were 49.057, 34.914, 33.547, 84.261 and 54.164 µg/mL, respectively) as compared to TAE (IC50 = 175.426 µg/mL) and aspirin (IC50 = 300.340 µg/mL), while the unbound compounds (palmatine and tetrahydropalmatine) had a very weak antiplatelet effect (IC50 > 200 µg/mL). DISCUSSION AND CONCLUSION: This study is the first reported work for antiplatelet components screening in Rhizoma Corydalis. Seven compounds were detected and identified by HPLC-DAD/LC-MS, of which five platelet-targeted compounds were discovered.

Prediction of placental barrier permeability: a model based on partial least squares variable selection procedure.

Assessing the human placental barrier permeability of drugs is very important to guarantee drug safety during pregnancy. Quantitative structure-activity relationship (QSAR) method was used as an effective assessing tool for the placental transfer study of drugs, while in vitro human placental perfusion is the most widely used method. In this study, the partial least squares (PLS) variable selection and modeling procedure was used to pick out optimal descriptors from a pool of 620 descriptors of 65 compounds and to simultaneously develop a QSAR model between the descriptors and the placental barrier permeability expressed by the clearance indices (CI). The model was subjected to internal validation by cross-validation and y-randomization and to external validation by predicting CI values of 19 compounds. It was shown that the model developed is robust and has a good predictive potential (r2=0.9064, RMSE=0.09, q2=0.7323, rp2=0.7656, RMSP=0.14). The mechanistic interpretation of the final model was given by the high variable importance in projection values of descriptors. Using PLS procedure, we can rapidly and effectively select optimal descriptors and thus construct a model with good stability and predictability. This analysis can provide an effective tool for the high-throughput screening of the placental barrier permeability of drugs.

Research progress of pharmacological activities and analytical methods for plant origin proteins.

As one of the important active components of traditional Chinese medicine (TCM), plant origin active proteins have many significant pharmacological functions. According to researches on the plant origin active proteins reported in recent years, pharmacological effects include anti-tumor, immune regulation, anti-oxidant, anti-pathogeny microorganism, anti-thrombus, as well as hypolipidemic and hypoglycemic activities of plant origin were reviewed, respectively. On the other hand, the analytical methods including chromatography, spectroscopy, electrophoresis and mass spectrometry for plant origin proteins analysis were also summarized. The main purpose of this paper is providing a reference for future development and application of plant active proteins.

[Applications of platelets in studies on traditional Chinese medicines promoting blood circulation to remove blood stasis].

Thrombotic diseases in different forms become a great threat to human health. Such anti-platelet aggregation drugs as aspirin and clopidogrel are common drugs in clinic. However, along with the appearance of resistance and side effects of western anti-platelet aggregation drugs, anti-platelet aggregation traditional Chinese medicines promoting blood circulation to remove blood stasis have gradually become an important study orientation. Platelet is one of major participant in thrombosis, and plays an important role as a bioactive material in studies on traditional Chinese medicines promoting blood circulation to remove blood stasis, mainly involving two aspects--the evaluation for the anti-platelet aggregation activity of traditional Chinese medicines and the screening of their active components. This paper summarized the applications of platelets in studies on traditional Chinese medicines promoting blood circulation to remove blood stasis, so as to provide basis for further studies.

Development of a cell-based high-throughput peroxisome proliferator-activated receptors (PPARs) screening model and its application for evaluation of the extracts from Rhizoma Coptis.

To date, peroxisome proliferator-activated receptors (PPARs) are becoming the new therapeutic targets for the treatment of metabolic diseases, such as Type 2 diabetes, obesity, and cardiovascular disease. In this study, a cell-based high-throughput PPARs (PPARα/ß/γ) model was developed for the screening of PPARs agonists. The screening conditions were evaluated through analyzing the expression value of luciferase. Finally, 24 h of drug acting time, 5 times of the dilution factor of luciferase zymolyte, and about 2 × 10(4) cells/ well on HeLa cells in 96-well plates were used, respectively. Furthermore, the quality of high-throughput screening (HTS) in stability and reliability was evaluated by the Z'-factor. Additionally, different extracts of Rhizoma Coptis and berberine were tested by the developed method. The results suggested that both the EtOAc extract and berberine were able to activate PPARα/ß/γ, and Rhizoma Coptis contains potential natural agonists of PPARs besides berberine. In conclusion, the developed HTS assay is a simple, rapid, stable, and specific method for the screening of PPARs natural agonists.

[Research progress on the construction and applications of metal-organic frameworks in chromatographic stationary phases].

Metal-organic frameworks (MOFs) are a class of porous crystalline materials composed of metal centers or clusters assembled with organic ligands. These materials possess excellent properties, such as large surface areas, high porosities, uniform pore sizes, and diverse structures. Thus, MOFs have been widely applied in various fields, including catalysis, adsorption, sensing, sample pretreatment, and chromatographic separation. The applications of MOFs as stationary phases for chromatographic separation and analysis have attracted considerable attention from the research community in recent years. Compared with traditional chromatographic stationary phases, such as mesoporous silica, nanoparticles, and porous layers, MOFs possess flexible and tunable pore sizes and structures, thereby enabling precise control over their intermolecular interactions. Furthermore, the wide range of functional ligands and topologies of MOFs could potentially facilitate the separation and analysis of complex samples. These unique advantages render MOFs highly suitable for constructing novel chromatographic stationary phases.This article focuses primarily on the construction methods of MOFs as chromatographic stationary phases, and provides an overview of the latest research advancements in their applications in several chromatographic separation techniques such as high performance liquid chromatography (HPLC), gas chromatography (GC), and capillary electrochromatography (CEC). The existing methods for the preparation and construction of MOFs-based chromatographic stationary phases are classified and evaluated. The construction methods for MOFs as stationary phases for HPLC mainly include filling, precursor-doped polymerization, and post-modification. The construction methods for MOFs as stationary phases for GC predominantly include in situ growth, static coating, and dynamic coating. The stationary phases for CEC can be categorized into packed columns, monolithic columns, and open-tubular columns. Compared with monolithic and packed columns, open-tubular CEC (OT-CEC) offers numerous advantages, including a more flexible and convenient preparation method, enhanced compatibility with various separation media, and higher separation efficiency. Consequently, OT-CEC has emerged as an important method for investigating the preparation of stationary phases for CEC. Several methods such as physical adsorption, covalent attachment, and electrostatic interactions have been developed for the preparation and modification of MOFs-based CEC stationary phases, and extensive studies have been conducted to optimize the performance and applications of MOFs in OT-CEC. However, the existing methods for constructing MOFs-based chromatographic stationary phases present certain limitations. Therefore, the selection of the appropriate MOFs, optimization of their preparation methods, and examination of their performance in different separation modes have become the focus of intensive research.This review also summarizes the different analytical targets (e. g., chiral small molecules, biomacromolecules, and nonchiral molecules) and corresponding separation effects achieved using various MOFs-based chromatographic stationary phases. Finally, future studies focusing on the development of MOFs as chromatographic separation media are discussed. Overall, this review provides a valuable reference for the rational construction and practical applications of advanced MOFs-based chromatographic stationary phases.

Glucagon-like peptide 1 receptor plays a critical role in geniposide-regulated insulin secretion in INS-1 cells.

AIM: To explore the role of the glucagon-like peptide 1 receptor (GLP-1R) in geniposide regulated insulin secretion in rat INS-1 insulinoma cells. METHODS: Rat INS-1 insulinoma cells were cultured. The content of insulin in the culture medium was measured with ELISA assay. GLP-1R gene in INS-1 cells was knocked down with shRNA interference. The level of GLP-1R protein in INS-1 cells was measured with Western blotting. RESULTS: Geniposide (0.01-100 µmol/L) increased insulin secretion from INS-1 cells in a concentration-dependent manner. Geniposide (10 µmol/L) enhanced acute insulin secretion in response to both the low (5.5 mmol/L) and moderately high levels (11 mmol/L) of glucose. Blockade of GLP-1R with the GLP-1R antagonist exendin (9-39) (200 nmol/L) or knock-down of GLP-1R with shRNA interference in INS-1 cells decreased the effect of geniposide (10 µmol/L) on insulin secretion stimulated by glucose (5.5 mmol/L). CONCLUSION: Geniposide increases insulin secretion through glucagon-like peptide 1 receptors in rat INS-1 insulinoma cells.

Synergistic recognition of transferrin by using performance dual epitope imprinted polymers.

Transferrin (Trf) is a new type of active drug targeting carrier and disease biomarker that regulates the balance of iron ions in human body. The recognition and isolation of Trf is of great significance for disease diagnosis and treatment. Thus, a new type of magnetic dual affinity epitope molecularly imprinted polymer coated on Fe3O4 nanoparticles (Fe3O4@DEMIP) was successfully prepared for specific recognition of Trf. C-terminal nonapeptide and Trf glycan were selected as bi-epitope templates for metal chelation and boron affinity immobilization, respectively. 4-vinylphenylboric acid (4-VP), N-isopropyl acrylamide (NIPAM) and zinc acrylic were used as functional monomers. Results showed that Fe3O4@DEMIP exhibited excellent specific recognition ability adsorption capacity toward Trf, with an adsorption of 43.96 mg g-1 (RSD = 3.28%) and a more satisfactory imprinting factor (about 6.60) than that of other reported imprinting methods. In addition, Fe3O4@DEMIP displayed pH, temperature and magnetic sensitivity properties to realize temperature and pH-controlled recognition and release of target proteins and magnetic rapid separation. Furthermore, the Fe3O4@DEMIP coupled with high-performance liquid chromatography (HPLC) analysis was successfully used for specific recognition of Trf in biosamples. This study provides a reliable protocol for preparing metal chelation and boron affinity dual affinity bi-epitope molecularly imprinted polymers for synergistic and efficient recognition of biomacromolecules in the complex biological systems.

Selective separation and inexpensive purification of paclitaxel based on molecularly imprinted polymers modified with ternary deep eutectic solvents.

Paclitaxel (PTX) is a powerful anticancer natural product, with its separation and purification having been widely studied. In this work, new molecular imprinted polymers (MIPs) using deep eutectic solvents (DESs) with different molar ratios were prepared as functional monomers. These were then used as adsorbents in solid phase extraction (SPE) for the separation of PTX from its structural analogs. The polymers were characterized by energy disperive X-rays (EDX), scanning electron microscopy (SEM), thermogravimetric analysis (TGA) and fourier transform infrared spectroscopy (FT-IR). The results suggested that the formative regular DES-MIPs had an even pore-size distribution and a large specific surface area. The dynamic adsorption and static adsorption showed that the DES-MIPs had excellent adsorption performance, with a maximum adsorption capacity and optimum adsorption time of 87.08 mg/g and 180 min, respectively. The selective adsorption experiments showed that the material had outstanding selectivity, and the maximum selectivity factor was 6.20. For stability, after six consecutive adsorption and desorption cycles, the DES-MIPs maintained the perfect stability and reusability. Furthermore, the fabricated SPE column was successfully utilized for extracting and eluting PTX. This study provides a reliable protocol for the separation and purification PTX from its structural analogs and the DES-MIPs materials have excellent potential application value in pharmaceutical industry.

[Applications of real-time monitoring techniques in chemical synthetic ingredients].

Real-time monitoring technique for process parameters and/or insight variables of chemical synthetic ingredients is a novel chemical process analysis method, which can real time monitor the chemical synthetic ingredients, reveal the mechanism of chemical reaction occurring, reaction courses and kinetic characteristics, and monitor, control and adjust chemical reaction to determine the endpoint of reaction and enhance selectivity of reaction, quality and yields of product. Many real-time monitoring techniques were achieved to satisfy the demands in several chemical synthetic reactions. The structure and principles of current real-time monitoring techniques was stated, and a review was summarized on its applications in chemical synthetic ingredients. The research, development and applications of real-time monitoring techniques such as spectrometry (i. e. ultraviolet-visible spectrophotometry, infrared spectrometry, Raman spectroscopy, nuclear magnetic resonance spectroscopy, mass spectrometry and fluorescence spectroscopy), chromatography (i. e. thin layer chromatography, gas chromatography, high performance liquid chromatography and capillary electrophoresis) and their coupled techniques (i. e. GC-MS, GC-IR and LC-MS) for chemical synthetic ingredients were evaluated. The coupled techniques were utilized to take the advantages of their high performance separation and quantitative power of chromatography, and sensitive and qualitative identification capacity of spectrometric techniques could realize the real-time monitoring for special chemical synthetic ingredients in complex systems. The future developmental trends and application prospects of real-time monitoring techniques are also discussed. With the research & development of microprocessor and embedded system, the real-time monitoring instrument for chemical synthetic ingredients will have a trend to miniaturization, intelligence, digitization, functionalization and multichannel with widely versatile and strongly compatible features.