A novel method for the detection of hydrogen peroxide in aqueous solution was developed via reaction between H₂O₂, trivalent titanium ion (Ti³⁺) and 4-(2-thiazolylazo) resorcinol (TAR), resulting in a ternary complex with a maximum UV absorbance at 530 nm. The CE detection of H₂O₂ was fast, sensitive and cost-effective without pretreatment procedures. H₂O₂ was detected within 15 min at 1 to 100 µM range with the lowest detection limit at 1.0 µM. Under the optimized CE conditions, the concentration of H₂O₂ in coffee or tea extract was quantitatively determined. Our results show that CE detection of the ternary complex of H₂O₂-Ti³⁺-TAR has potential applications for the detection of H2O2 in aqueous sources.
Capillaries* ; Coffee ; Electrophoresis, Capillary* ; Hydrogen Peroxide* ; Hydrogen* ; Limit of Detection ; Methods ; Tea ; Titanium*

OBJECTIVELower respiratory tract infections continue to pose a significant threat to human health. It is important to accurately and rapidly detect respiratory bacteria. To compensate for the limits of current respiratory bacteria detection methods, we developed a combination of multiplex polymerase chain reaction (PCR) and capillary electrophoresis (MPCE) assay to detect thirteen bacterial pathogens responsible for lower respiratory tract infections, including Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Mycoplasma pneumoniae, Legionella spp., Bordetella pertussis, Mycobacterium tuberculosis complex, Corynebacterium diphtheriae, and Streptococcus pyogenes.

METHODSThree multiplex PCR reactions were built, and the products were analyzed by capillary electrophoresis using the high-throughput DNA analyzer. The specificity of the MPCE assay was examined and the detection limit was evaluated using DNA samples from each bacterial strain and the simulative samples of each strain. This assay was further evaluated using 152 clinical specimens and compared with real-time PCR reactions. For this assay, three nested-multiplex-PCRs were used to detect these clinical specimens.

RESULTSThe detection limits of the MPCE assay for the 13 pathogens were very low and ranged from 10-7 to 10-2 ng/μL. Furthermore, analysis of the 152 clinical specimens yielded a specificity ranging from 96.5%-100.0%, and a sensitivity of 100.0% for the 13 pathogens.

CONCLUSIONThis study revealed that the MPCE assay is a rapid, reliable, and high-throughput method with high specificity and sensitivity. This assay has great potential in the molecular epidemiological survey of respiratory pathogens.

Bacteria ; classification ; genetics ; isolation & purification ; Bacteriological Techniques ; DNA, Bacterial ; genetics ; Electrophoresis, Capillary ; methods ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Respiratory Tract Infections ; microbiology ; Sensitivity and Specificity

OBJECTIVE: To establish the rapid PCR amplification program and system and to verify the technical indexes. METHODS: PCR multiplex and capillary electrophoresis detection of 24 autosomal STR loci and one Y-STR loci using the 6-color fluorescence marking technology, as well as A melogenin and Y-InDel. Meanwhile, sensitivity, specificity, identity, stability, mixing and a batch of sample tests were investigated, and the genotype of various routine samples and degraded, exfoliated cell samples were observed. RESULTS: The sensitivity of the system was 0.062 5 ng. In addition, the genotype could be detected accurately only around 65 min via rapid amplification. The species-specificity was high and the genotyping of all kinds of dry blood specimens of filter paper and mixed, degraded, exfoliated cell samples were accurate. CONCLUSION The rapid amplification system can significantly improve the detection rate, and obtain accurate and stable genotyping results, which may be important implications for the establishment of STR database and study on population genetics and forensic identification.
Electrophoresis, Capillary ; Fluorescence ; Genetics, Population ; Genotype ; Humans ; Microsatellite Repeats ; Multiplex Polymerase Chain Reaction/methods* ; Sensitivity and Specificity

OBJECTIVETo determine enantiomeric impurity in levocetirizine tablets by using capillary electrophoresis.

METHODSThe effects of pH and the concentrations of sulfated-Β-cyclodextrin (S-Β-CD) and buffer salt on chiral resolution were examined with S-Β-CD as chiral selector.

RESULTSA good enantioseparation of cetirizine was achieved with 30 mmol/L NaH2PO4 buffer solution (pH 7.0) containing 20 g/L of S-Β-CD.

CONCLUSIONThe method developed in the study is sensitive and reliable for determination of enantiomeric impurity in levocetirizine tablets.

Cetirizine ; analysis ; Electrophoresis, Capillary ; methods ; Stereoisomerism ; Tablets

OBJECTIVETo develop a capillary electrophoresis system for enantiomeric impurity test of repaglinide.

METHODSAn uncoated fused silica capillary (50 μm×50 cm, with an effective length of 41 cm) was used. The running buffer was composed of 30 mmol/L NaH2PO4 and 5 mg/ml carboxymethyl-β-cyclodextrin(pH 3.5).

RESULTSLinear range was 2.00-80.00 μg/ml (correlation coefficient was 0.9993). The average recovery rate was 92.5% to 105.0%.

CONCLUSIONThe method is simple, accurate and sensitive and it can be used for determination of enantiomeric impurities in repaglinide tablet.

Carbamates ; analysis ; Electrophoresis, Capillary ; methods ; Piperidines ; analysis ; Stereoisomerism ; Tablets

OBJECTIVE: To develop a multiplex allele-specific PCR (AS-PCR) assay with three-color fluorescence labeling for mitochondrial DNA (mtDNA) SNP typing. METHODS: Based on the principle of AS-PCR, the primer sets were designed for 20 SNP located on the coding region of mtDNA and divided into 2 groups labeled with FAM and HEX fluorescence, respectively. A primer set included two forward (reverse) allelic specific primers with different sizes and a generic reverse (forward) primer. Blood samples from 200 unrelated individuals were analyzed by AS-PCR and capillary electrophoresis. Three random samples at least for each SNP site were examined and verified by direct sequencing. The haplotype frequency was investigated. RESULTS: Distinct electropherograms of 200 blood samples were obtained successfully. The typing results of direct sequencing were identical to those obtained from AS-PCR. The minimum detectable DNA concentration was 0.2 pg under the system of 10 microL. The sensitivity of the DNA concentrations ranged from 0.5 to 5 pg. The 200 individuals were assigned into 15 haplotype, and the haplotype diversity was 0.906 0. CONCLUSION AS-PCR is a simple, rapid and efficient method for mtDNA SNP typing, and can be applied to forensic practice.
Alleles ; DNA ; DNA Primers ; DNA, Mitochondrial/analysis* ; Electrophoresis, Capillary ; Haplotypes ; Humans ; Mitochondria ; Polymerase Chain Reaction/methods* ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

OBJECTIVE: To assess the patterns of linkage disequilibrium (LD) of 16 STR loci on X chromo- some and investigate the genetic stability. METHODS: Genomic DNA samples extracted from blood stains from 500 unrelated individuals and 885 lineage members from Eastern Chinese Han population were genotyped through multiplex amplification using IDtyperX-16 kit by our independent research followed by capillary electrophoresis. LD was assessed by PowerMarker v3.25 software and mutation rate of every locus was analyzed. RESULTS: LD were not found at the 16 X-STR loci. Allele mutations were observed at 10 loci. Among them, mutation rates of DXS6809 and DXS7132 were both up to 0.0048. CONCLUSION When the 16 X-STR loci included in IDtyperX-16 kit were used for parentage testing, product princi- ples can be applied to calculate the likelihood, but mutation should be taken into consideration in the case that the genotypes do not meet the genetic law (especially at DXS6809 and DXS7132).
Alleles ; Asian People/genetics* ; Blood Stains ; China ; Chromosomes, Human, X/genetics* ; Electrophoresis, Capillary ; Female ; Forensic Genetics/methods* ; Gene Frequency ; Genetic Loci/genetics* ; Genotype ; Humans ; Linkage Disequilibrium/genetics* ; Microsatellite Repeats/genetics* ; Multiplex Polymerase Chain Reaction ; Mutation ; Mutation Rate

OBJECTIVETo establish fingerprints for Poecilobdella manillensis from Guangxi province using the high performance capillary electrophoresis (HPCE) method.

METHODElectrophoresis was performed on a fused silica capillary column (75 microm x 56 cm), with 25 mmol x L(-1) Na2HPO4-120 mmol x L(-1) Tris-16 mmol x L(-1) SDS (adjusted to pH 12.0 with 1 mol x L(-1) NaOH ) as the running buffer. The applied voltage was 17 kV, the temperature was 25 degrees C and the detection wavelength was 254 nm. The sample's hydrodynamic injection was 3.4 kPa x 6s and the duration was 27 min.

RESULTHPCE fingerprint was established with 13 common peaks. The similarity between fingerprints of P. manillensis in 10 batches and control fingerprints was more than 0.98.

CONCLUSIONThe method is so precise, reproducible and stable that it could be used as a new means for the quality control of P. manillensis.

Animals ; China ; Electrophoresis, Capillary ; methods ; Leeches ; chemistry ; Medicine, Chinese Traditional ; standards ; Quality Control ; Reproducibility of Results

OBJECTIVETo establish a capillary electrophoresis-mass spectrometry(CE-MS) method for the analysis of nineteen organonitrogen pesticides in Paeoniae Radix Alba.

METHODSCE-MS analysis was performed on a 70 cm X 50 μm fused-silica capillary. The optimal buffer was composed of 1 % formic acid and 15 % methanol(V/V, pH 2.2). The temperature of capillary was controlled at 25 degree. The separation voltage was +20 kV. The optimal MS parameters were as follows: ESI-MS analysis was performed in the positive mode; 90 % methanol containing 0.2 % formic acid with a flow rate of 8 μl·min(-1) was selected as the sheath liquid; the flow rate and temperature of drying gas were 6 L·min(-1) and 250 degree, respectively; The nebulizing gas pressure was set at 5 psig; The optimal values of fragmentor and ESI voltage were 100 V and 5 000 V, respectively.

RESULTSThe nineteen pesticides had good linearity over the testing ranges. The average recoveries were in the range of 80.1 %-108.4 % with RSDs less than 20 % (except ethoxyquin and spiroxamine, those of which were 29.2 % and 22.3 % at 0.01 mg.kg(-1) concentration level). The LODs of nineteen pesticides were 0.503 ≊10.1 μg.kg(-1).

CONCLUSIONThe method can be used effectively to analyze the nineteen organonitrogen pesticides residue in Paeoniae Radix Alba.

Electrophoresis, Capillary ; methods ; Mass Spectrometry ; methods ; Paeonia ; chemistry ; Pesticide Residues ; analysis

Caspases play important roles in cell apoptosis. Measurement of the dynamics of caspase activation in tumor cells not only facilitates understanding of the molecular mechanisms of apoptosis but also contributes to the development, screening, and evaluation of anticancer drugs that target apoptotic pathways. The fluorescence resonance energy transfer (FRET) technique provides a valuable approach for defining the dynamics of apoptosis with high spatio-temporal resolution. However, FRET generally functions in the single-cell level and becomes ineffective when applied in the high throughput detection of caspase activation. In the current study, a FRET sensor was combined with capillary electrophoresis (CE) to achieve a high throughput method for cellular caspase detection. The FRET-based CE system is composed of a homemade CE system and a laser source for detecting the dynamics of caspase-3 in various cells expressing sensors of caspase-3 that have been treated with anticancer drugs, such as cell cycle-independent drug cisplatin and specific cell cycle drugs camptothecin and etoposide, as well as their combination with tumor necrosis factor (TNF). A positive correlation between the caspase-3 activation velocity and drug concentration was observed when the cells were treated with cisplatin, but cells induced by camptothecin and etoposide did not show any apparent correlation with their concentrations. Moreover, different types of cells presented distinct sensitivities under the same drug treatment, and the combination treatment of TNF and anticancer drugs significantly accelerated the caspase-3 activation process. Its high throughput capability and detection sensitivity make the FRET-based CE system a useful tool for investigating the mechanisms of anticancer drugs and anticancer drug screening.
Antineoplastic Agents ; pharmacology ; Camptothecin ; pharmacology ; Caspase 3 ; metabolism ; Cisplatin ; pharmacology ; Drug Screening Assays, Antitumor ; Electrophoresis, Capillary ; methods ; Enzyme Activation ; drug effects ; Etoposide ; pharmacology ; HeLa Cells ; Hep G2 Cells ; Humans ; Neoplasms ; enzymology ; pathology ; Time Factors ; Tumor Necrosis Factor-alpha ; pharmacology