Qionglin Liang

Carbon nanotube-modified electrodes for the simultaneous determination of dopamine and ascorbic acid

The voltammetric separation of dopamine and ascorbic acid was studied with cyclic voltammetry at two kinds of carbon nanotube-modified electrodes (coated and intercalated). The anodic peak difference reached 270 mV under the present conditions. The separation mechanism and effect factors were carefully studied. Using various types of surfactants as coating dispersants of carbon nanotubes, it was demonstrated that the charge nature of the surfactants had a strong effect on the electrochemical behavior of dopamine and ascorbic acid. When the oxidation solution of carbon nanotubes was changed from the most commonly used mixed concentrated nitric acid and sulfuric acid (1 + 3 v/v) to dilute nitric acid and to hydrochloric acid, the anodic peak separation value of dopamine and ascorbic acid increased significantly, and it was shown that carboxylic acid groups attached to the carbon nanotubes were an adverse factor for the discrimination of DA from AA. These results indicated that the resolution of DA and AA was mainly attributable to the stereo porous interfacial layer formed from aggregated pores and inner cavities of the carbon nanotubes. The modified electrodes exhibited an attractive ability to measure DA and AA simultaneously and showed good stability and reproducibility.

Carbon nanotube-intercalated graphite electrodes for simultaneous determination of dopamine and serotonin in the presence of ascorbic acid

The modified electrode was constructed by intercalating carbon nanotubes (CNT) on graphite surface, which exhibits an attractive ability to determine dopamine (DA) and serotonin (5-HT) simultaneously in the presence of ascorbic acid (AA), resulting in a high DPV current response to 5-HT at 0.32 V and DA at 0.13 V and a favorably low response to AA at −0.07 V versus SCE in a 50 mM phosphate buffer (pH 7.4). The properties and behaviors of the modified electrode were characterized using CV and DPV. The results show that the high sensitivity and selectivity is attributed mainly to the unique carbon surface of the nanotubes and the porous interfacial layers that come from the subtle tubule structure of CNT. The linear calibration range for DA in the presence of 0.5 mM AA and 5 μM 5-HT was 0.5–10 μM, with a correlation coefficient of 0.9996, and a detection limit of 0.1 μM. A linear relationship was found for 5-HT in the range of 1–15 μM containing 5 μM DA and 0.5 mM AA, with a correlation coefficient of 0.9998 and a detection limit of 0.2 μM. The current sensitivities were 1.89 and 8.05 μA μM−1 for 5-HT and DA, respectively. Rabbits brain homogenate presented no interference to 5-HT and DA simultaneous detection.

Crystal structure of the Caenorhabditis elegans apoptosome reveals an octameric assembly of CED-4.

The CED-4 homo-oligomer or apoptosome is required for initiation of programmed cell death in Caenorhabditis elegans by facilitating autocatalytic activation of the CED-3 caspase zymogen. How the CED-4 apoptosome assembles and activates CED-3 remains enigmatic. Here we report the crystal structure of the complete CED-4 apoptosome and show that it consists of eight CED-4 molecules, organized as a tetramer of an asymmetric dimer via a previously unreported interface among AAA(+) ATPases. These eight CED-4 molecules form a funnel-shaped structure. The mature CED-3 protease is monomeric in solution and forms an active holoenzyme with the CED-4 apoptosome, within which the protease activity of CED-3 is markedly stimulated. Unexpectedly, the octameric CED-4 apoptosome appears to bind only two, not eight, molecules of mature CED-3. The structure of the CED-4 apoptosome reveals shared principles for the NB-ARC family of AAA(+) ATPases and suggests a mechanism for the activation of CED-3.

Simultaneous determination and quantification of seven major phospholipid classes in human blood using normal-phase liquid chromatography coupled with electrospray mass spectrometry and the application in diabetes nephropathy.

A rapid and specific analytical method for simultaneous determination and quantification of seven major phospholipid classes in human blood was developed by normal-phase high-performance liquid chromatography tandem mass spectrometry. The optimal separation was achieved by using mobile phase hexane (A) and 2-propanol with water, formic acid and ammonia as modifiers (B) using an HPLC diol column. Isocratic elution method was used for better repeatability and no balance time. The seven major phospholipid classes in human blood that were detected including phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI) phosphatidylcholine (PC), lysophosphatidylcholine (Lyso-PC), and sphingomyelin (SM). That can be separated in this condition. Every phospholipid class contains many molecular species which have similar structure. The structure of phospholipids molecular species was identified by ion-trap MS(n) which produced ion fragments. And the qualification was completed by TOF-MS which shows good accuracy. Through the accurate quantification of one representative phospholipids molecule in each class, a method for simultaneous estimation hundreds of molecular species in seven major classes was established. The intra-day and inter-day precision and recovery had been investigated in detail. The RSD of precision for most compound is below 8% and RE is below 10%. Recovery is almost over 80%. This method was applied to phospholipids disorder related with diabetes nephropathy successfully. The concentrations of most phospholipids for normal people are higher than that for diabetic nephropathy (DN) patients in three phases. For most of phospholipids, with the development of DN the concentration was decreasing.

Metabonomic Study of Chinese Medicine Shuanglong Formula as an Effective Treatment for Myocardial Infarction in Rats

A UPLC/TOF-MS-based metabonomic study was conducted to assess the holistic efficacy of Traditional Chinese Medicine Shuanglong Formula (SLF) for myocardial infarction in rats. Thirty male Sprague-Dawley rats were randomly divided into five groups after surgery. The Panax ginseng group, Salvia miltiorrhiza group, and SLF group were treated with water extractions of Panax ginseng (PG), Salvia miltiorrhiza (SM), and SLF (the ratio of SM to PG was 3:7) at a dose of 5 g/kg·w·d for 21 consecutive days, respectively; the model group and sham surgery group were both treated with 0.9% saline solution. Urinary samples for metabonomic study, serum samples for biochemical measurement, and heart samples for histopathology were collected. As a result, metabonomics-based findings such as the PCA and PLS-DA plotting of metabolic state and analysis of potential biomarkers in urine correlated well to the assessment of serum biochemistry and histopathological assay, confirming that SLF exerted synergistic therapeutic efficacies to exhibit better effect on MI compared to PG or SM. The shifts in urinary TCA cycle as well as pentose phosphate pathway suggested that SLF may diminish cardiac injury of MI with its potential pharmacological effect in the regulation of myocardial energy metabolism.

On-chip manipulation of continuous picoliter-volume superparamagnetic droplets using a magnetic force

A microfluidic device for generating monodisperse superparamagnetic droplets and rapidly manipulating desired droplets into designated sub-microchannels by an external magnetic force is described. Superparamagnetic magnetite (Fe3O4) nanoparticles are synthesized by a chemical co-precipitation method. They are well dispersed in the water-phase to form a superparamagnetic fluid that is sheared into picoliter-volume monodisperse superparamagnetic droplets by the oil-phase in a T-junction PDMS microchannel. Superparamagnetic droplets always flow into sub-microchannel 1 due only to laminar flow without a magnetic field. But they are deflected from the direction of laminar flow by a perpendicular magnetic field. The results show that the deflection is proportional to the magnetic field gradient and magnetic nanoparticle concentration, and it is closely related to the magnet position. The observed experimental results make a good match with theoretical analysis. Single or bulk superparamagnetic droplets are successfully manipulated into the designated sub-microchannels 2 and 3 respectively, only by changing the positions of the magnet. Relatively high efficiency is obtained with more than 10 superparamagnetic droplets precisely manipulated per second. This simple and robust apparatus has wide applications in high throughput drug delivery/screening, immunoassay, cell research and synthesis of magnetic microparticles due to good biological compatibility and monodispersity of superparamagnetic droplets.

One-step synthesis of magnetic graphene oxide nanocomposite and its application in magnetic solid phase extraction of heavy metal ions from biological samples.

A novel magnetic graphene oxide nanocomposite was synthesized by one-step coprecipitation method and characterized by transmission electron microscopy, X-ray photoelectron spectroscopy and vibrating sample magnetometer. The nanocomposite beard many intriguing properties, including chemical stability, high adsorption capacity, and superparamagnetic. These properties evoked great interest and desire of its exploration in magnetic solid-phase extraction of heavy metal ions from complex samples. Several parameters effecting the analytical performance, such as the sample pH, amounts of adsorbent, sample volumes, elution volumes, and coexisting ions, had been investigated in detail. The adsorbed metal ions were easy eluted by controlling the pH condition and the materials could be reused more than 20 times. Under the optimized conditions, the limits of detection were 0.016, 0.046, 0.395, 0.038, 0.157 μg L(-1) for Co(2+), Ni(2+), Cu(2+), Cd(2+), and Pb(2+), respectively. The intra-day relative standard deviations (n=5) were in the range of 1.8-5.5% at 10 μg L(-1). The proposed method was successfully applied to biological sample analysis and got excellent recoveries in the range of 81-113% even the matrix was complex.

Rapid qualitative and quantitative analyses of Asian ginseng in adulterated American ginseng preparations by UPLC/Q-TOF-MS.

A new method using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS) was developed for the rapid qualitative and quantitative analyses of Asian ginseng (Panax ginseng C.A.Meyer) in adulterated American ginseng (Panax quinquefolium L.) preparations within 2min. The method was based on the baseline chromatographic separation of isomeric compounds of ginsenoside Rf and 24(R)-pseudoginsenoside F(11), two potential chemical markers present in Panax ginseng C.A.Meyer and P. quinquefolium L. methanolic extracts. The chromatographic separation was achieved by UPLC, which used a column with 1.7microm particle packing which enabled the higher peak capacity, greater resolution, increased sensitivity and higher speed of analysis. Ginsenoside Rf and 24(R)-pseudoginsenoside F(11) were separated on baseline with retention times of 1.5 and 1.7min, respectively. Ginsenoside Rf and 24(R)-pseudoginsenoside F(11) were identified and conformed unambiguously by accurate mass measurement and their different fragmentation pathways were performed on Q-TOF-MS. Quantitative analysis was carried out under selective ion monitoring (SIM) mode. The limit of detection (LOD) of this UPLC/Q-TOF-MS analysis for ginsenoside Rf and 24(R)-pseudoginsenoside F(11) was 0.05 and 0.08ng, respectively. Ginsenoside Rf was linear over the range of 0.164-16.4ng with a correlation coefficient (R(2)) of 0.9997, while 24(R)-pseudoginsenoside F(11) was linear from 0.243 to 24.3ng with an R(2) of 0.9989. Furthermore, inter-day and intra-day precisions were obtained below 4.0% and the analytical method was fully validated. 12 batches of self-prepared adulterated samples, 11 batches of Asian ginseng, 16 batches of American ginseng and 13 batches of commercial American ginseng preparations were tested. The method developed is rapid, accurate, reliable and highly sensitive for qualitative and quantitative analyses of Asian ginseng and American ginseng.

Phospholipidomic identification of potential plasma biomarkers associated with type 2 diabetes mellitus and diabetic nephropathy

Type 2 diabetes mellitus (T2DM) and its attendant complications, such as diabetic nephropathy (DN), impose a significant societal and economic burden. The investigation of discovering potential biomarkers for T2DM and DN will facilitate the prediction and prevention of diabetes. Phospholipids (PLs) and their metabolisms are closely allied to nosogenesis and aggravation of T2DM and DN. The aim of this study is to characterize the human plasma phospholipids in T2DM and DN to identify potential biomarkers of T2DM and DN. Normal phase liquid chromatography coupled with time of flight mass spectrometry (NPLC-TOF/MS) was applied to the plasma phospholipids metabolic profiling of T2DM and DN. The plasma samples from control (n=30), T2DM subjects (n=30), and DN subjects (n=52) were collected and analyzed. The significant difference in metabolic profiling was observed between healthy control group and DM group as well as between control group and DN group by the help of partial least squares discriminant analysis (PLS-DA). PLS-DA and one-way analysis of variance (ANOVA) were successfully used to screen out potential biomarkers from complex mass spectrometry data. The identification of molecular components of potential biomarkers was performed on Ion trap-MS/MS. An external standard method was applied to quantitative analysis of potential biomarkers. As a result, 18 compounds in 7 PL classes with significant regulation in patients compared with healthy controls were regarded as potential biomarkers for T2DM or DN. Among them, 3 DM-specific biomarkers, 8 DN-specific biomarkers and 7 common biomarkers to DM and DN were identified. Ultimately, 2 novel biomarkers, i.e., PI C18:0/22:6 and SM dC18:0/20:2, can be used to discriminate healthy individuals, T2DM cases and DN cases from each other group.

Plasma esterified and non-esterified fatty acids metabolic profiling using gas chromatography–mass spectrometry and its application in the study of diabetic mellitus and diabetic nephropathy

Using gas chromatography-mass spectrometry (GC-MS), a new metabolic profiling method was established to assess the levels of non-esterified fatty acids (NEFAs) and esterified fatty acids (EFAs) in plasma. The extraction method was simple and robust without removing protein process. With this method 25 fatty acids (FAs), both EFAs and NEFAs, can be recognized simultaneously with only 10 μL plasma. 15 of the 25 can be precisely quantified. The method was validated and then applied into clinical metabonomics research. Five clinical groups including 150 cases were involved. The relationship between FA levels and diabetic mellitus (DM) as well as diabetic nephropathy (DN) pathology was speculated. Furthermore, the possible pathological causes and effects were discussed in detail. Potential biomarkers (p value <0.01) were screened with Students t-test. With the application of partial least squares-discriminant analysis (PLS-DA), different stages were distinguished. The result may be useful for the pathology study of metabolic syndromes, and may also be helpful for monitoring the progression of DM and DN.