Simultaneous determination of four active compounds in Centella asiatica by supramolecular solvent-based extraction coupled with high performance liquid chromatography-tandem mass spectrometry.

In the present study, a new and rapid method for determining four bioactive compounds of Centella asiatica (C. asiatica) in herbs was developed based on high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). Supramolecular solvent (SUPRAS), formed by n-hexanol, tetrahydrofuran (THF) and water, was used for extracting madecassoside (MS), asiaticoside (AS), asiatic acid (AA) and madecassic acid (MA) from herbs. The sample was extracted with 4 mL of SUPRAS for 5 min. Then centrifugation was performed for phase separation followed by direct analysis by HPLC-MS/MS. Driving forces for the extraction of herbs in the SUPRAS involved both dispersion and hydrogen bond interactions. The effect of the parameters, including compounds of supramolecular solvents, dosage and vortex time, on the extraction efficiency was investigated. The recoveries were carried out at three levels with spiked samples and in the range of 91.6%-99.9%, with relative standard deviations between 1.7%-7.9%. The novel SUPRAS method, coupled with HPLC-MS/MS, was proved to be efficiency, green, and sensitive. It was applied for determination of four target compounds in herbs.

Separation and identification of cetirizine enantiomers in human urine by capillary electrophoresis and circular dichroism independent of their standards.

Enantioseparation and determination of chiral drugs are of vital importance in biochemical and pharmaceutical research due to the different biological activity, mechanism, and toxicity of individual enantiomers. As a second-generation H(1)-antagonist, cetirizine's pharmaceutical activity is mainly derived from the levocetirizine while the dextro-enantiomer is ineffective and even associated with side effects. Herein, the enantiomers of cetirizine were separated by capillary electrophoresis and identified by electronic circular dichroism. Satisfactory linear relationship was found between the circular dichroism signal at λmax and the electrophoretic peak area difference in the nonracemic mixture of enantiomers. It made possible identification and quantification of cetirizine enantiomers independent of single enantiomer standards. The method's feasibility was demonstrated on the enantiomeric excess experiments of oral drugs measured in human blank urine. Additionally, the separation and determination of cetirizine in human urine after administration were also realized by capillary electrophoresis, indicating the method was sensitive enough for pharmacokinetic study.

[Determination of 19 illegally added chemical ingredients in hair loss prevention cosmetics by ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry].

Cosmetic products for hair loss prevention are often mixed with prohibited substances such as hormones, antibiotics, and forbidden pharmacologically active substances. Although drugs increase the efficacy of cosmetic products, they cause skin irritation and allergic reactions, upon long-term exposure. Given the increasing number of hair loss prevention cosmetics on the market, the need to guarantee product safety calls for efficient and reliable methods to identify illegal ingredients in these products. Chromatography combined with high-resolution mass spectrometry offers the advantages of high resolution and high throughout, thus being a powerful technique for simultaneously detecting illegal ingredients in cosmetics. In this study, an ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS) method for detecting 19 illegal chemical components was established. Combined with the scientific database, a screening platform for hair loss prevention cosmetics was constructed. The effect of extraction solvent was investigated. The chromatographic and mass spectrometry conditions were optimized. Under the optimal conditions, separation was achieved within 20 min on an ACQUITY UPLC BEH C18 column (100 mm×2.1 mm, 1.7 µm). Acetonitrile and 2 mmol/L ammonium formate solution containing 0.05% formic acid were used as mobile phases for gradient elution. The 19 compounds were detected by positive ion electrospray ionization (ESI) in the MSE mode. The chromatographic retention time, precursor ions, product ions of the target analytes, and abundance ratio were collected to construct a screening database with UNIFI software. The 19 compounds were well separated, with good linearity. The limits of detection (LODs) and limits of quantification (LOQs) were 0.025-0.05 µg/g and 0.075-0.15 µg/g, respectively. Hair lotion and shampoo, which are commonly marketed as hair loss prevention cosmetics, were selected as the respective matrices for the recovery experiment. The average recoveries of the 19 compounds ranged from 68.6% to 118.6%, and the relative standard deviations (RSDs) were 0.3%-10.3%. Then, 77 batches of cosmetic samples were detected and screened under the same conditions. The TOF-MS information, including the retention time, ion addition mode, mass-to-charge ratio of the parent ions and fragment ions, as well as the abundance ratio, were compared between the cosmetic samples and the standard MS information with UNIFI software. Finally, two batches of samples that were illegally adulterated with minoxidil and finasteride were identified. The ESI fragmentation pathway of the product ions from minoxidil was also proposed. The matrix matching external standard method was used to determine the amounts of minoxidil and finasteride in the two batches of hair lotion, and they were as high as 60 mg/g and 0.31 mg/g, respectively. This result revealed that multiple chemical components were simultaneously added to hair loss prevention cosmetics. Furthermore, the amount of the illegally added drug was very high, indicating high safety risk for consumers using such cosmetics. The present method has the advantages of simple operation, high sensitivity, and good reproducibility. It can be used for rapid screening and simultaneous quantitative analysis of various illegal chemicals in hair loss prevention cosmetics.

Application of the Mechanical High-Pressure Method Combined with High-Performance Liquid Chromatography-Tandem Mass Spectrometry for Determination of Veterinary Drug Residues in Incurred Chicken and Rabbit Muscle Tissues.

Rapid sample preparation is a key step in the field of food safety. A mechanical high-pressure method using a laboratory-made meat press machine was first introduced in this study to process the incurred muscle samples of chicken and rabbit. By applying high pressure to animal muscle, the meat juice was obtained. After extraction and purification, veterinary drug residues in the juice were qualitatively and quantitatively analyzed by using high-performance liquid chromatography-tandem mass spectrometry. The sample press conditions and extraction solvents were optimized. Under the optimal conditions, all veterinary drug residues, including tetracycline, enrofloxacin, clenbuterol, ampicillin, lincomycin, erythromycin, and sulfadiazine, in the incurred samples were detected. The residual concentration of drugs in samples obtained by using the mechanical high-pressure method can reach up to 94.0% of that obtained by using the common homogenization method, suggesting that drug residues exist in the tissue juice, which justifies the use of the mechanical high-pressure method. Moreover, with the mechanical high-pressure method, the sample preparation time was shortened by five times, and the consumption of the extraction solvent was reduced by 50%, relative to the homogenization method.

[Determination of 42 veterinary drug residues in common animal derived food by dispersive solid phase extraction coupled with liquid chromatography-tandem mass spectrometry].

A liquid chromatography-tandem mass spectrometry method, coupled with a dispersive solid phase extraction (DSPE) procedure for sample preparation, was developed to determine 13 classes of 42 veterinary drugs in four representative animal-derived foods. The analytes were dispersed with water, extracted with acetonitrile containing 5% (v/v) formic acid, salted out by salts, and purified by DSPE. The analytes were separated on a C18 column (100 mm×2.1 mm, 2 µ m) with gradient elution using the mobile phase containing methanol and 0.1% (v/v) formic acid aqueous solution. Electrospray ionization-mass spectrometry was performed in multiple reaction monitoring mode for analysis of the 42 compounds. The correlation coefficients of the standard calibration curves for the 42 veterinary drugs were all above 0.995. Most recoveries at three spiked levels in the four representative matrixes ranged from 65.8% to 135.5%, with relative standard deviations of 0.5%-14.2% (n=6). The limits of detection (LODs, S/N=3) and the limits of quantification (LOQs, S/N=10) were 0.01-1.68 µ g/kg and 0.01-5.62 µ g/kg, respectively. The method is simple, rapid, sensitive, and suitable for the simultaneous determination of the 42 veterinary drugs in animal-derived food.

A sensitive colorimetric method for the determination of nitrite in water supplies, meat and dairy products using ionic liquid-modified methyl red as a colour reagent.

This paper describes a colorimetric approach to determine trace amounts of nitrite in water supplies, meat and dairy products using 1-butyl-3-methylimidazolium-modified methyl red ([BMIM]MR) as a colour reagent. The technique capitalises on the catalytic effect of nitrite on the oxidative degradation of [BMIM]MR by potassium bromate in acidic media. The absorbances were proportional to nitrite concentrations in the range of 8.70×10(-2) to 4.17 µM with a detection limit of 1.64×10(-2) µM. Compared with the method using methyl red as a colour reagent, 60 times improvement of sensitivity was obtained. Activation energy and the apparent rate constant for the catalytic reaction are 61.11 kJ mol(-1) and 1.18×10(4) s(-1), respectively. The proposed method was successfully applied for the analysis of nitrite in Yellow River water, chicken, and milk with recoveries ranging from 96% to 105%.

In situ synthesis of MIL-100(Fe) in the capillary column for capillary electrochromatographic separation of small organic molecules.

Because of the unusual properties of the structure, the metal organic frameworks (MOFs) have received great interest in separation science. However, the most existing methods for the applications of MOFs in separation science require an off-line procedure to prepare the materials. Here, we report an in situ, layer-by-layer self-assembly approach to fabricate MIL-100(Fe) coated open tubular (OT) capillary columns for capillary electrochromatography. By a controllable manner, the OT capillary columns with a tailored MIL-100(Fe) coating have been successfully synthesized. The results of SEM, XRD, FT-IR, and ICP-AES indicated that MIL-100(Fe) was successfully grafted on the inner wall of the capillary. Some neutral, acidic and basic analytes were used to evaluate the performance of the MIL-100(Fe) coating OT capillary column. Because of the size selectivity of lattice aperture and hydrophobicity of the organic ligands, three types of analytes were well separated with this novel MIL-100(Fe) coating OT capillary column. For three consecutive runs, the intraday relative standard deviations (RSDs) of migration time and peak areas were 0.4-4.6% and 1.2-6.6%, respectively. The interday RSDs of migration time and peak areas were 0.6-8.0% and 2.2-9.5%, respectively. The column-to-column reproducibility of retention time was in range of 0.6-9.2%. Additionally, the 10 cycles OT capillary column (10-LC) could be used for more than 150 runs with no observable changes on the separation efficiency.

On-line capillary electrophoresis enrichment by combining chitosan trapping with surfactant assisted sample stacking for the ultratrace determination of organic acids in Plateau alfalfa roots.

In this paper, four organic acids constituents of Plateau alfalfa roots have been identified and detected by a novel capillary electrophoresis (CE) strategy which combined chitosan (CS) trapping and cetyltrimethyl ammonium bromide (CTAB) assisted sample stacking. Under the optimized condition, organic acids, i.e., aconitic acid, gallic acid, citric acid and l-malic acid were concentrated and separated within 3 min. Validation parameters of this method (such as detection limits, linearity and precision) were also investigated and the limit of detection (LOD) was 2.41-53.9 ng mL(-1). Linearity was obtained over the magnitude range of 5-4000 ng mL(-1) approximately for different organic acids and 3×10(2)-1.5×10(4) folds enrichment was achieved. The method has been applied to the determination of organic acids in roots of normal grown Plateau alfalfa and stressing affected Plateau alfalfa. Satisfactory results and recoveries were obtained in the analysis without costly and complicated sample pretreatment.

In situ loading of gold nanoparticles on Fe3O4@SiO2 magnetic nanocomposites and their high catalytic activity.

In this work, a facile approach was successfully developed for in situ catalyzing Au nanoparticles loaded on Fe3O4@SiO2 magnetic nanospheres via Sn(2+) linkage and reduction. After the Fe3O4@SiO2 MNPs were first prepared via a sol-gel process, only one step was needed to synthesize the Fe3O4@SiO2-Au magnetic nanocomposites (Fe3O4@SiO2-Au MNCs), so that both the synthesis step and the reaction cost were remarkably decreased. Significantly, the as-synthesized Fe3O4@SiO2-Au MNCs showed high performance in the catalytic reduction of 4-nitrophenol to 4-aminophenol and could be reused for several cycles with convenient magnetic separability. This approach provided a useful platform based on Fe3O4@SiO2 MNPs for the fabrication of Au or other noble metal magnetic nanocatalysts, which would be very useful in various catalytic reductions.

Preparation and characterization of open-tubular capillary column modified with graphene oxide nanosheets for the separation of small organic molecules.

A novel coated capillary was prepared by immobilizing graphene oxide (GO) on the fused-silica capillary (75 µm i.d.) which was derivatized by 3-aminopropyl-trimethoxysilane (APTMS). The bare capillary, APTMS modified capillary (NH2-capillary) and GO coated capillary (GO-capillary) were characterized by streaming potentials (SPs), fluorescence microscope and scanning electron microscope (SEM). The results indicated that the capillary was successfully modified with GO sheets via covalent bonding and electrostatic effect. Compared with bare capillary, greater separation efficiency was achieved by GO-capillary column as a result of the increasing interactions between the small organic molecules and the inner wall of the GO-capillary column originated from the π-π electrostatic stacking. For three consecutive runs, the intra-day relative standard deviations (RSDs) of migration time and peak areas were 0.6-4.3% and 2.8-9.3%, respectively. The inter-day relative standard deviations of migration time and peak areas were 0.2-8.3% and 4.5-9.6%. Additionally, one GO-capillary column could be used for more than 100 runs with no observable changes on the separation efficiency.

A new on-line concentration method of cationic molecules in capillary electrophoresis by a hyphenated micelle to solvent stacking coupling with large amount sample electrokinetic stacking injection.

In this paper, we established a new on-line method using micelle to solvent stacking (MSS) technique combining with large amount sample electrokinetic stacking injection (LASEKSI) for the analysis of cationic molecules. In this MSS-LASEKSI, by modulating the integral EOF across the capillary, a equilibrium state was formed and can be maintained for a long time, leading to the continuous stacking of the analytes on the basis of MSS. Thereby, an extremely large amount sample was permitted to be injected into the capillary and then an improved enrichment fold can be achieved comparing with the each case. The variables affecting the performance of MSS-LASEKSI were investigated and discussed. Under the optimized conditions, 6.3 × 10(3)- and 6.4 × 10(2)-fold enrichment in peak heights upon normal CZE method (injected at 0.5 psi for 3 s) and number of plates of 2.9 × 10(6) and 6.5 × 10(5) were attained for berberine and theophylline, respectively. The developed method described here may provide prospects for exploiting a new concentration technique to achieve higher enrichment factor.

The binding affinity of amino acid-protein: hydroxyproline binding site I on human serum albumin.

The binding affinity between hydroxyproline (Hyp) and human serum albumin (HSA) was investigated under simulated physiological conditions, using molecular modeling in combination with steady-state fluorescence, synchronous fluorescence, time-resolved fluorescence, UV-vis absorption, circular dichroism (CD), and Fourier transform infrared (FT-IR) spectroscopy. Molecular modeling studies suggested that the Hyp molecule was situated within subdomain IIA of HSA. The fluorescence quenching analysis indicated that the fluorescence of HSA was quenched by Hyp with a dynamic quenching mechanism. The binding constants were calculated according to Scatchard's equation and implied that Hyp can bind to different binding sites on HSA. The thermodynamic analysis implied that hydrophobic forces were the main interaction in the Hyp-HSA system, which was found to be in line with the results of molecular modeling. Furthermore, the conformational structure of HSA was changed with various amounts of Hyp, which was confirmed by synchronous fluorescence, UV-vis absorption, CD, and FT-IR spectra.

Protein adsorption on citrate modified magnetic nanoparticles.

Magnetic iron oxide nanoparticles were prepared by chemical co-precipitation method and then modified with sodium citrate. These iron oxide nanoparticles were characterized by transmission electron microscopy (TEM), X-ray diffractometer (XRD), and vibrating sample magnetometer (VSM). BSA was adsorbed on these citrate modified nanoparticles using two types of buffers (acetate buffer, pH 4.0, 4.7 and phosphate buffer, pH 7.4). The results showed that the maximum adsorption of BSA was 83 mg/g at its isoelectric point (pH 4.7). Fourier-transform infrared spectroscopy (FTIR) was used to confirm the attachment of citrate groups and BSA on the prepared magnetic nanoparticles. BSA was desorbed from nanoparticles under alkaline conditions, which was confirmed by SDS-PAGE electrophoresis, UV-vis and fluorescence spectra. The desorbed BSA showed small changes in its structure. The adsorption results indicated that BSA adsorption on citrate modified iron oxide nanoparticles occurred mainly by electrostatic mechanism.

Graphene oxide-Fe3O4 magnetic nanocomposites with peroxidase-like activity for colorimetric detection of glucose.

In this report, graphene oxide-Fe(3)O(4) magnetic nanocomposites were demonstrated to possess intrinsic peroxidase-like activity and enhanced affinity toward H(2)O(2). The nanocomposites retain their magnetic properties and can be effectively separated by a magnet. Significantly, they were proved to be novel peroxidase mimetics which could quickly catalyze oxidation of the peroxidase substrate 3,3,5,5-tetramethylbenzidine (TMB) in the presence of H(2)O(2), producing a blue-colored solution. Kinetic analysis indicates that the catalytic behavior is in accord with typical Michaelis-Menten kinetics and follows a ping-pong mechanism. On the basis of the high activity, the reaction provides a simple, sensitive and selective method for colorimetric detection of glucose in diabetic urine.

Rapid and sensitive determination of hydroxyproline in dairy products using micellar electrokinetic chromatography with laser-induced fluorescence detection.

Many reports have focused on the determination of hydroxyproline (Hyp) in blood plasma, urine sample, meat and meat products, however, there are few concerned with the Hyp assay in dairy products for food quality assurance up to now. In this paper, we described a sensitive and automated approach for the determination of Hyp in milk powder, liquid milk, milk drink and soymilk powder samples by micellar electrokinetic chromatography (MEKC) based on in-capillary derivatization for the first time. Under the optimal conditions, derivatization and separation procedure could be completed within 7 min and the detection limit for Hyp was 1.6±0.5 ng mL(-1). Comparing with the existing alternatives, the present method exhibited some relevant advantages, including full automation, satisfactory sensitivity, and short analysis time for Hyp assay in dairy products.

[Recent advances in the application of high performance capillary electrophoresis for food safety].

In recent years, food safety incidents become a serious social problem. Foods are usually complex mixtures consisting of a large diversity of molecules. Analysis of foods is a topic that demands the development of rapid, robust, efficient, sensitive and cost-effective analytical methodologies. Therefore, new techniques for food safety purpose are required by analytical chemists. Capillary electrophoresis (CE) is a popular separation technique that possesses fast and efficient performances in an automated way with minimum consumption of sample and reagents. Nowadays, CE represents a desired strategy for the determination of many compounds or molecules in various kinds of food. In this paper, the review intends to provide the recent innovative developments reported in food safety analysis using CE methods for a full overview. As a fundamental review, it focuses on the introduction and detection of several common hazardous materials existing in food such as non-food additives, pesticide residues, veterinary drug residues, heavy metal ion contaminants, toxins, biphenol A and phthalates in packaging materials and so on. Furthermore, this review prospects the main development direction of CE in this field for the future. A total of 63 papers published during the period of Jan 2009 - Jun 2012 are included in the present review.

Preparation of guanidine group functionalized magnetic nanoparticles and the application in preconcentration and separation of acidic protein.

Guanidine group (Gnd) functionalized magnetic nanoparticles (Fe3O4@SiO2@NH2-Gnd) were synthesized and characterized in this work for the first time. The characterization of Fe3O4@SiO2@ NH2-Gnd nanoparticles was demonstrated by transmission electron microscopy, X-ray diffraction, Fourier transform infrared spectra, vibration sample magnetometer, and zeta potential analyzer. The novel multifunctional nanoparticles were served as a solid-phase extraction sorbent for easy isolation and preconcentration of acidic protein from aqueous solution only using a magnet. Bovine serum albumin (BSA) was selected as a model protein and the main experimental parameters influencing the adsorption and desorption efficiency were investigated and optimized. Under the optimum conditions, the particles reached saturated adsorption within 20 min and exhibited significant specific recognition for the acidic proteins. Fifteen fold enrichment efficiency was achieved and the detection limits was 45 ng x mL(-1) for BSA by capillary electrophoresis (CE). The practical application of the novel nanoparticles as a sorbent for the isolation and preconcentration of acidic proteins from basic proteins was demonstrated by effective separation and enrichment of bovine serum albumin from lysozyme and cytochrome C mixture, which was assayed by CE.

In-line preconcentration of oxidized and reduced glutathione in capillary zone electrophoresis using transient isotachophoresis under strong counter-electroosmotic flow.

Transient isotachophoresis (tITP) can improve the sensitivity of capillary electrophoresis (CE). In general, it was carried out under the condition of suppressed electroosmotic flow (EOF). However, some special conditions, such as extreme low pH background electrolyte and coating were needed to achieve the requirements of suppressed EOF. In this work, an approach of tITP under the strong counter-EOF in open system (counter-EOF-tITP) is presented for the rapid and sensitive preconcentrating the reduced glutathione (GSH) and the oxidized glutathione (GSSG) without modifying the capillary and the commercial CE instrument. The parameters of the experimental system, such as the concentration of leading electrolyte, the injected amount of terminating electrolyte and the injected pressure of sample were investigated in detail to understand the mechanism of counter-EOF-tITP. The sensitivity enhancement factors were of 320 for GSH and 280 for GSSG. In addition, the detection limit of 23.4 and 18.0 microg L(-1) for GSH and GSSG was achieved, respectively. The method's applicability was demonstrated by determining GSH and GSSG in tomato and human serum.