Detection and quantification of chitosan aggregates by pressure-assisted capillary zone electrophoresis

An approach to the detection and determination of chitosan aggregates in acetic acid solutions is proposed using pressure-assisted capillary zone electrophoresis. Processes of chitosan aggregation are studied depending on the composition of dispersion medium and storage time. The presence of several species of positively charged chitosan aggregates is revealed for the first time. Particle sizes in the range 20–2500 nm are determined by scanning electron microscopy and static and dynamic light scattering. The dependence of the shape of electropherograms on particle size distribution obtained under the same conditions is found. A trend to changing electrophoretic mobility depending on the size of the aggregate is observed, which enables the approximate evaluation of the polydispersity of chitosan solutions. Chitosan is used for the effective dynamic modification of capillaries, which does not require the introduction of a modifier into the background electrolyte.     

Identification and quantification of natural isoxazolinone compounds by capillary zone electrophoresis.

A capillary zone electrophoresis (CZE) method that is specific, simple, rapid and also cheap was developed to analyse some natural UV-absorbing isoxazolinone compounds with toxic potential present in legume seedlings. The six most common natural isoxazolinone compounds were separated within 10 min with 25 mM potassium phosphate (pH 7.5) containing 8% 1-propanol as running buffer. A 60 cm coated fused-silica capillary (52.6 cm effective length x 75 microm I.D.), with an electric field of 375 V/cm at 30 degrees C was used. The limit of detection ranged from 0.01 mM (3.0 microg/ml) to 0.03 mM (7.7 microg/ml). Linearity between peak areas and concentrations ranging from 0.05 mM to 1.75 mM were determined for each isoxazolinone. The correlation coefficient was 0.9954 or greater. Both relative migration time and peak area were reproducible. The RSD of relative migration time is between 0.44 and 1.94% and RSD of peak area is between 1.26 and 6.86%. The concentrations of isoxazolinones in Lathyrus odoratus and L. sativus seedlings obtained by CZE were in agreement with the previous results from HPLC.     

A new method of quantification of pipemidic-acid by capillary zone electrophoresis with end-column amperometric detection

Capillary zone electrophoresis was employed for the determination of pipemidic acid using an end-column amperometric detection with a carbon fiber microdisk array electrode, at a constant potential of -1.10 V vs. saturated calomel electrode. The optimum conditions of separation and detection were 1.2 x 10(-4) mol/LNaOAc - 8.8 x 10(-4) mol/ LHOAc for the buffer solution, 20 kV for the separation voltage, 5 kV and 10 s for the injection voltage and the injection time. The limit of detection was 1.05 x 10(-7) mol/L or 189 amol (S/N=3). The relative standard deviation was 0.31% for the migration time and 2.0% for the electrophoretic peak current. The method was applied to determining pipemidic acid in human serum.     

Separation of nicotine metabolites by capillary zone electrophoresis and capillary zone electrophoresis/mass spectrometry

The use of capillary zone electrophoresis (CZE) and capillary zone electrophoresis/mass spectrometry (CZE/MS) has been demonstrated, in principle, for the separation of nicotine and nicotine metabolites. The buffer system developed for separation and detection by CZE/UV was modified for use in CZE/MS analysis. Several of the metabolites are isobaric and tandem mass spectrometric (MS/MS) techniques have been used to differentiate such analytes.     

Rapid quantification of total genomic DNA methylation degree by short-end injection capillary zone electrophoresis

We propose a new capillary zone electrophoresis method applying short-end injection technique for the fast evaluation of methylcystosine/total cytosine ratio after acidic DNA hydrolysis. By short-end injection and by using a 100 mmol/l Tris solution titrated with 1 mol/l phosphoric acid to pH 3.75 as background electrolyte, cytosine and methylcytosine were separated with a good resolution in less than 1.5 min. Stepwise multiple linear regression with DNA methylation degree as the dependent variable and age, cysteine, homocysteine and methionine as independent variables, showed a negative association with age and that total cysteine is the most important determinant of DNA methylation.     

Separation of nitrophenols by capillary zone electrophoresis

Separation conditions suitable to a rapid resolution of a group of eight nitrophenols by capillary zone electrophoresis (CZE) were found. Required differences in their effective mobilities were achieved via host-guest complexation of -cyclodextrin combined with intermolecular interactions involved by polyvinylpyrrolidone. When both additives were present in the carrier electrolyte at pH=9.1 nitrophenols could be separated in the column of a, 300 m I.D. and 180 mm in the length within 8–9 minutes. It is shown that the column of such an I.D. providing enhanced sample load capacity, can operate with high separation efficiencies as maintaining zone dispersions due to Joule heating on a tolerable level. CZE on-line coupled with isotachophoretic sample pretreatment is shown to provide the concentration limits of detection at low ppb concentrations by using an on-column photometric detector operating at 254 and 405 nm detection wavelengths.     

毛细管区带电泳法测定粉针剂中头孢拉定的含量

用毛细管区带电泳法测定头孢拉定的含量 ,未涂层毛细管柱 (75 μm×48.5cm ,有效长度 40cm) ,电压 2 8kV ,检测波长 2 3 0nm ,温度 2 0℃ ,进样 5×1 0 3Pa× 3s。运行缓冲液为 2 5mmol/L硼砂缓冲液。方法的线性范围 3 1 .2 2μg/mL～ 749.2 8μg/mL ,检测限为 1 .1 7μg/mL。     

A rapid urine creatinine assay by capillary zone electrophoresis.

Using capillary zone electrophoresis, the urine creatinine (uCr) assay was validated in extemporaneous diluted urine, both in healthy subjects and athletes, with the uCr concentration as a reference value to compare excretion rates of other metabolites in the same samples. The electrokinetic sample injection was carried out at 10 kV per 10 s; UV absorbance detection was at 254 nm. Using standard samples, the creatinine migration mean time in 100 mmol/L acetate buffer, pH 4.4, was 3.3+/-0.2 min; the repeatability for absolute migration mean time was 0.6% and peak height repeatability was 2.9%. The correlation coefficient of the standard curve was r = 0.999 and the detection limit was 23.1 micromol/L. Intra- and interassay coefficients of variation (CV) were 3.0 and 3.6%, respectively; recovery was 99+/-3% and linearity was r= 0.98. Normal urine samples were diluted 1:80 in run buffer. The present CE urine creatinine assay showed a good correlation with HPLC and with Jaffe methods (r = 0.98 and r = 0.97, respectively; p < 0.0001). The uCr in the morning urine samples of 34 healthy males (M), 38 healthy females (F), and 83 male athletes (A) was 10.4+/-6.1 mmol/L, 10.8+/-8.1 mmol/L and 13.2+/-6.5 mmol/L, respectively. The uCr difference (p < 0.02) between M and A and a correlation (p < 0.05) with age in A were observed.     

Stacking in capillary zone electrophoresis

Due to the short light path of the capillaries, the CE detection limit based on concentration, is far less than that of HPLC and not sufficient for many practical applications. Several methods, based on different electrophoretic maneuvers, can concentrate the sample (stack) easily on the capillary before the separation step of capillary zone electrophoresis (CZE). These methods incorporate different types of discontinuous buffers as the means for invoking different velocities to the same analyte molecules to produce a sharpening of the band (stacking). In CZE, these buffers can be often very simple such as sample dilution or adding to the sample a high concentration of a fast mobility ion. However, in other applications these buffers can be as complicated as those required for isotachophoresis. Stacking can often yield a concentration factor of 5-30-fold, which can improve greatly in CZE the detection limits bringing them very close to those of HPLC. Different methods of stacking, the importance of discontinuous buffers and the different mechanism for concentration on the capillary are reviewed here. As there is a need for more practical applications, there will be more methods devised for stacking in CZE.     

Cocaine detection by structure-switch aptamer-based capillary zone electrophoresis

Aptamers, which are nucleic acid oligonucleotides that can bind targets with high affinity and specificity, have been widely applied as affinity probes in capillary electrophoresis (CE). Due to relative weak interaction between aptamers and small molecules, the application of aptamer-based CE is still limited in certain compounds. A new strategy that is based on the aptamer structure-switch concept was designed for small molecule detection by a novel CE method. A carboxyfluorescein (fluorescein amidite, FAM) label DNA aptamer was first incubated with partial complementary strand (CS), and then the free aptamer and the aptamer-CS duplex were well separated and determined by metal cation mediated CE/laser-induced fluorescence. When the target was introduced into the incubated sample, the hybridized form was destabilized, resulting in the changes of the fluorescence intensities of the free aptamer and the aptamer-CS duplex. The length of CS was investigated and 12 mer CS showed the best sensitivity for the detection of cocaine. The presented CE-LIF method, which combines the separation power of CE with the specificity of interactions occurring between target, aptamer, and CS, could be a universal detection strategy for other aptamer-specified small molecules.     

Stability study of somatropin by capillary zone electrophoresis

Human Growth Hormone (hGH) is a monomeric 22 kilo Dalton (kDa), 191 amino acid protein with an isoelectric point (pI) close to pH 5, produced in the anterior pituitary gland. High level production of somatropin (recombinant hGH) is done in Escherichia coli (E. coli) to meet the demand and to avoid possible Creutzfeldt-Jacob disease (CJD).The present study was initiated on the basis of results from post-marketing control of all somatropin preparations on the Norwegian market. The samples consisted of preparations presented as somatropin in solution, and some freeze-dried preparations were also included for comparison.The present study showed a significant degree of degradation of somatropin in solution. Deamidation increased over time for preparations in solution, as well as for freeze-dried preparations after dissolution. Preparations in solution showed high content of deamidated and cleaved forms. Freeze-dried preparations after dissolution and storage showed high content of deamidated forms, but low content of cleaved forms. Also, in one preparation, an unknown peak was detected in the electropherogram from capillary zone electrophoresis (CZE), eluting after the principal peak, in front of the Gln-18 somatropin peak.     

Determination of aristolochic acid by capillary zone electrophoresis

A simple and rapid capillary zone electrophoresis (CZE) method for the determination of aristolochic acid (AA) in dietary supplements and selected herbs is described. A clear separation of AA from other sample constituents was achieved within 5 minutes without any sample clean up. A mixture of 20 mM-morpholinethanesulphonic acid+10 mM-BisTrisPropane+0.2% hydroxyethylcelullose in 10% methanol serves as a background electrolyte. The linearity, accuracy, intra-assay and detection limit of the developed method are 200–6000 ng/mL, 95–103%, 3.5%, and 50 ng/ml, respectively. Ease of use, sufficient sensitivity and low running cost are the most important attributes of the CZE method. The proposed CZE method was compared with HPLC.     

Separation of human fibrinopeptides by capillary zone electrophoresis

We describe a method for the simultaneous determination of the five fibrinopeptide forms derived from the thrombin-promoted activation of human fibrinogen by capillary zone electrophoresis (CZE). The fibrinopeptide mixture was first desalted by a solid-phase extraction (SPE) step. The analysis was performed in reversed polarity in a highly cross-linked polyethylene glycol (PEG)-coated capillary with UV-light absorption detection at 200 nm. Several parameters including buffer concentration and pH, presence of an organic modifier, temperature, and applied voltage, have been tested. The best separations were obtained within 20 min, utilizing a 20 mM sodium phosphate buffer without organic modifier, in the narrow 6.1-6.2 pH range, at 25 degrees C, with an applied voltage of 20 kV. Quantitative analysis is made possible by the use of sheep fibrinopeptide A as an internal standard to correct for both extraction and injection errors.     

毛细管区带电泳法拆分手性药物环扁桃酯

近年来,随着不同种类的手性添加剂[1]在毛细管电泳(CZE)中的使用,毛细管电泳越来越显示出其强有力的手性拆分性能。具有特殊笼状结构并含有多个手性中心的环糊精及其衍生物是毛细电泳手性分离研究中最常采用的手性添加添[2-4]。本文合成了环糊精衍生物单3 O 苯基胺甲酰基 β CD[2]并以之作为手性选择剂分离了β CD及手性药物环扁桃酯。1　实验部分932 3 HVPS高压电源(山东省化工研究院),DD 2000型可调波长紫外检测器(中国科学院大连化学物理研究所),XWT型记录仪(上海大华仪表厂),pHS 25型酸度计(上海雷磁仪器厂),石英毛细管45cm…     

Determination of dyes in foodstuffs by capillary zone electrophoresis

A rapid method based on capillary zone electrophoresis coupled with photodiode-array detection has been developed to determine the dyes Tartrazine E-102, Sunset Yellow FCF E110, Amaranth E-123, New Coccine E-124, Patent Blue V calcium salt E-131 and Allura Red AC E-129 in foodstuffs. Separation was done by using a Bare CElect-FS75 CE column, using a 10 mM phosphate buffer at pH 11.0. Hydrodynamic injections at 0.5 p.s.i. for 4 s (21 nl of sample) and 20 kV separation voltage were used. The quantitation limits for the six dyes ranged from 3 to 6 microg/ml. A linear relationship between 3 to 95 microg/ml, with correlation coefficient better than 0.995 was obtained. This method has been applied to the determination of the studied dyes in beverages, jellies and syrups.     

Enantiomeric separation of racemic clenbuterol by capillary zone electrophoresis

A method for capillary electrophoretic enantiomeric separation of a racemic clenbuterol has been established with hydroxypropyl-β-cyclodextrin as the chiral selector. General equations and data analysis are presented to relate mobility to the equilibrium constants in simple binding equilibria and used to determine binding constants and thermodynamic parameters for host-guest complexation of clenbuterol enantiomers with hydroxypropyl-β-cyclodextrin as a selector. The effects of β-cyclodextrin type and concentration, buffer type, concentration and pH, as well as separation voltage and capillary temperature were investigated in detail. A maximal resolution of 6.78 was obtained. The binding constants of the host-guest complex of clenbuterol enantiomers with hydroxypropyl-β-cyclodextrin, K _R-CD and K _S-CD are 22.50 and 43.09 l mol^-1, respectively.     

Analysis of illicit amphetamine seizures by capillary zone electrophoresis

Capillary zone electrophoresis was applied for the determination of amphetamine and related substances in seized drugs. A buffer made of 0.1 M phosphoric acid adjusted to pH 3.0 with triethanolamine was selected. With this background electrolyte, triethanolamine is adsorbed to the capillary wall and the electroosmotic flow is reversed. This gives rise to peaks with good symmetry, high efficiency and reproducible migration times. The separation of the different analytes was performed in a fused-silica capillary thermostatted at 25 degrees C and the applied voltage was 25 kV. Under these experimental conditions, amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyethamphetamine, N-methyl-1-(1,3-benzodioxol-5-yl)-2-butamine and ephedrine were resolved within 8 min and without interference from adulterants usually found in illicit powders. Their identification by the migration time was confirmed by their UV spectra recorded with a diode array UV detector (190-350 nm). The selected method was then applied to identify these substances in illicit tablets known as "Ecstasy" and the MDMA determined in these samples according to a laboratory validation procedure.     

Split-flow injector for capillary zone electrophoresis

A simple construction of a split-flow injector eliminating some common problems connected with the use of such devices is described. It consists of a low-pressure pump, an injection valve and a delivery tube in which the separating capillary inlet is fixed. The sample is injected without moving the separating capillary inlet and without interrupting the applied voltage. The grounded electrophoretic electrode is close to the injection valve so that all metal parts of the injector are kept at a sufficiently low potential. Minimum length and small internal diameter of delivery tube minimizes additional sample zone broadening. The effects of some experimental parameters, such as the position of the separation capillary inlet with respect to the background solution flow direction and background solution flow-rate are experimentally studied. The injector was tested primarily for the electrokinetic injection.