Zhining Xia

Direct UV detection of underivatized amino acids using capillary electrophoresis with online sweeping enrichment

This is an original report proposed a CE method for direct analysis of the underivatized amino acids using UV detection with relatively higher sensitivity, which was based on coordination interactions between amino acids and Cu (II) ions. In addition, an online sweeping preconcentration technique was easily combined to improve the detection sensitivity. Satisfying separations of the amino acids were obtained under optimized conditions: 50 mmol/L CuSO4-0.05% HAc-H2O (pH 4.5), and the separation voltage of 15 kV. The LODs for the analytes ranged from 0.1 to 0.5 micromol/L. The linearity of detection for all analytes was two orders of magnitude with the correlation coefficients greater than 0.99. The repeatability was displayed with an RSD less than 3% for migration time and peak height (n = 5). Moreover, some amino acids in real samples of human saliva and green tea were analyzed by this direct UV detection CE method with acceptable sensitivity.

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.

A facile and versatile approach for controlling electroosmotic flow in capillary electrophoresis via mussel inspired polydopamine/polyethyleneimine co-deposition.

Electroosmotic flow (EOF), which reveals the charge property of capillary inner surface, has an important impact on the separation performance and reproducibility of capillary electrophoresis (CE). In this study, a novel, facile and versatile method to achieve diverse and controllable EOF in CE was reported based on the co-deposition of mussel-inspired polydopamine (PDA) and branched polyethyleneimine (PEI) on the capillary inner surface as the hybrid functional coating. After these PDA/PEI co-deposited columns were reinforced by the post-incubation of FeCl3, various magnitude and direction of EOF in CE could be easily achieved by varying a number of preparation parameters, including the mass ratio of DA/PEI and the molecular weight of PEI (including PEI-600, PEI-1800, PEI-10000 and PEI-70000). The separation effectiveness and stability of the hybrid coated columns were verified by the analysis of aromatic acids and aniline derivatives. The results showed that the controllable and diverse EOF was important in enhancing the separation performance of the analytes. The baseline separation of all the five aromatic acids can be achieved in 7 min with high separation efficiency by using the PDA/PEI-600 co-deposited column with the mass ratio of 6:1. On the other hand, with the PDA/PEI-70000 co-deposited column with the mass ratio of 6:1, the aniline compounds were easily baseline separated within 10 min. By contrast, using the bare and PDA coated columns, the migration of the aromatic acids was very slow and the baseline separation of the aniline compounds cannot be obtained. Moreover, the co-deposited columns showed long lifetime and good stability. The relative standard deviations for intra-day, inter-day and capillary-to-capillary repeatability of the PDA/PEI-600 co-deposited column with the mass ratio of 6:1, which was reinforced by the post-incubation of FeCl3, were all lower than 5%.

Fractionation of Volatile Constituents from Curcuma Rhizome by Preparative Gas Chromatography

A preparative gas chromatography (pGC) method was developed for the separation of volatile components from the methanol extract of Curcuma rhizome. The compounds were separated on a stainless steel column packed with 10% OV-101 (3 m × 6 mm, i.d.), and then, the effluent was split into two gas flows. One percent of the effluent passed to the flame ionization detector (FID) for detection and the remaining 99% were directed to the fraction collector. Five volatile compounds were collected from the methanol extract of Curcuma rhizome (5 g/mL) after 83 single injections (20 uL) with the yield of 5.1–46.2 mg. Furthermore, the structures of the obtained compounds were identified as β-elemene, curzerene, curzerenone, curcumenol, and curcumenone by MS and NMR spectra, respectively.

Determination of eight isoflavones in Radix Puerariae by capillary zone electrophoresis with an ionic liquid as an additive

Isoflavones are the main active components in Radix Puerariae, the dry root of Pueraria lobata or P. thomsonii. We developed a simple capillary zone electrophoresis (CZE) method with an ionic liquid (IL) as an additive for the simultaneous determination of eight isoflavones – ononin, daidzin, genistin, biochanin A, formononetin, puerarin, genistein and daizein – in Radix Puerariae. The experimental conditions, including the concentration of sodium tetraborate, the pH, the type and concentration of the ILs, the applied voltage and the capillary temperature, were investigated. The eight analytes (detection wavelength 260 nm) were well separated within 9 min using a running buffer of 30 mM sodium tetraborate and 50 mM 1-butyl-3-methylimidazolium tetrafluoroborate as an additive at pH 9.5, with an applied voltage of 18 kV and a capillary temperature of 25 °C. The method developed was fully validated (limit of detection, 1.72–4.92 μg mL−1; limit of quantification, 3.64–9.84 μg mL−1; intra-day precision, 1.1–4.7% RSD; inter-day precision, 2.1–6.6% RSD; and recovery, 93.1–107.5% with 4.0–5.9% RSD) and was successfully applied to the determination of the eight analytes in three Radix Puerariae samples. The results indicated that P. lobata contains more isoflavones than P. thomsonii. CZE with an IL as an additive is a promising method for the analysis of natural products.

Improvement of microemulsion electrokinetic chromatography for measuring octanol-water partition coefficients.

Microemulsion electrokinetic chromatography (MEEKC) has been used to indirectly measure octanol–water partition coefficients (log Pow) of compounds. In order to obtain an accurate log Pow value, the electrophoretic mobilities of the microemulsion phase (μme) and the analyte (μeff) in MEEKC must be accurately required. However, in conventional MEEKC, the shortage of obtaining μme with a tracing method was discovered, and the influences of concentration, injection volume of analyte, and high electric field on measuring μeff were also found. In this paper, a novel method called improved MEEKC (I‐MEEKC) was developed to avoid the problems mentioned above. In I‐MEEKC, a nonlinearity fitting program was used to obtain μme to avoid the error from tracing μme; the extrapolating method was used to eliminate the effects of concentrations and injection volumes of analytes on μeff measurement, and an enough stable microemulsion was selected to eliminate the effect of high electric field on μeff measurement. Then the novel method was applied to estimate log Pow of uncharged compounds and charged pharmaceuticals compared to the conventional MEEKC. The log Pow of all analytes obtained by I‐MEEKC agreed with those obtained by classical shake flask or literature values, the errors between them were within 0.1 logarithm units, better than the ones by conventional MEEKC.

Natural Products for Antithrombosis

Thrombosis is considered to be closely related to several diseases such as atherosclerosis, ischemic heart disease and stroke, as well as rheumatoid arthritis, hyperuricemia, and various inflammatory conditions. More and more studies have been focused on understanding the mechanism of molecular and cellular basis of thrombus formation as well as preventing thrombosis for the treatment of thrombotic diseases. In reality, there is considerable interest in the role of natural products and their bioactive components in the prevention and treatment of thrombosis related disorders. This paper briefly describes the mechanisms of thrombus formation on three aspects, including coagulation system, platelet activation, and aggregation, and change of blood flow conditions. Furthermore, the natural products for antithrombosis by anticoagulation, antiplatelet aggregation, and fibrinolysis were summarized, respectively.

Enantioselective analysis of ofloxacin enantiomers by partial‐filling capillary electrophoresis with bacteria as chiral selectors

The enantiomeric separation of ofloxacin enantiomers (OFLX) was achieved by using capillary electrophoresis partial-filled with Escherichia coli, Pseudomonas aeruginosa (Gram-negative), and Staphylococcus aureus (Gram-positive) as chiral selectors. Experimental parameters, including the concentration of background electrolyte, applied voltage, length of the filled bacteria plug, and pH of the buffer, were intensively investigated. Baseline separation of OFLX could be achieved within 7 min by using E. coli and P. aeruginosa as chiral selectors under the following conditions: electrophoretic buffer composed of 10 mM phosphate buffer at pH 7.4, applied voltage at 15 kV, and the bacteria (6.0 × 10(8) cells/mL) were injected into the capillary by gravity with injection height of 17.5 cm for 180 s (E. coli), 300 s (P. aeruginosa), and 300 s (S. aureus), respectively. E. coli and P. aeruginosa had better chiral selectivity for OFLX than S. aureus, which was in good agreement with OFLX having better antimicrobial activity on Gram-negative rather than Gram-positive bacteria. A novel method was developed for the enantioselective separation of enantiomers using bacteria as chiral selectors, which provides a new approach for antimicrobials enantioselective analysis, chiral pharmacodynamics, and chiral pharmacokinetics studies.

Separation of cis- and trans-Asarone from Acorus tatarinowii by Preparative Gas Chromatography

A preparative gas chromatography (pGC) method was developed for the separation of isomers (cis- and trans-asarone) from essential oil of Acorus tatarinowii. The oil was primarily fractionated by silica gel chromatography using different ratios of petroleum ether and ethyl acetate as gradient elution solvents. And then the fraction that contains mixture of the isomers was further separated by pGC. The compounds were separated on a stainless steel column packed with 10% OV-101 (3 m × 6 mm, i.d.), and then the effluent was split into two gas flows. One percent of the effluent passed to the flame ionization detector (FID) for detection and the remaining 99% was directed to the fraction collector. Two isomers were collected after 90 single injections (5 uL) with the yield of 178 mg and 82 mg, respectively. Furthermore, the structures of the obtained compounds were identified as cis- and trans-asarone by 1H- and 13C-NMR spectra, respectively.

As-prepared MoS2 quantum dot as a facile fluorescent probe for long-term tracing of live cells.

Recently, the newly emerged two-dimensional nanomaterials, layered transition metal dichalcogenide (e.g. MoS2) nanosheets, have drawn tremendous attentions due to their extraordinary electronic and optical properties, and MoS2 quantum dots (MoS2 QDs) with lateral sizes less than 10 nm have been found to be highly luminescent. In the present study, a facile approach for large-scale preparation of MoS2 QDs by Na intercalation reaction without using any toxic organic reagents is proposed. MoS2 QDs were carefully characterized by various techniques including transmission electron microscopy, atomic force microscopy, dynamic light scattering, spectroscopy, in vitro cytotoxicology, and capillary electrophoresis. The as-prepared MoS2 QDs were strongly fluorescent, highly photo-stable, low in cytotoxicity, and readily reactive to thiols. These inherent properties of MoS2 QDs make them excellent fluorescent probes for long-term live cell tracing. The results of live cells imaging indicated that MoS2 QD stained cells remained highly fluorescent after long-term culture, and could be easily traced from other co-cultured cell lines.