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Biomedical subjects

D D Chen

Publications and source records attributed to D D Chen.

At least 19 recordsLinked to original sources
PubMed2001

Separation of free amino acids in human plasma by capillary electrophoresis with laser induced fluorescence: potential for emergency diagnosis of inborn errors of metabolism.

Free amino acids (AAs) in human plasma are derivatized with 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) and analyzed by capillary electrophoresis (CE) with laser induced fluorescence (LIF) detection. The labeling procedure is significantly improved over results reported previously. Derivatization can be completed in 40 min, with concentrations as low as 4 x 10(-8) M successfully labeled in favourable cases. Twenty-nine AAs (including 2 internal standards) are identified and can be reproducibly separated in 70 min. Migration time RSD values for 23 of these AAs were calculated and found in the range from 0.5 to 4%. The rapid derivatization procedure and the resolution obtained in the separation are sufficient for a semi-quantitative, emergency diagnosis of several inborn errors of metabolism (IEM). Amino acid profiles for both normal donor plasma samples and plasma samples of patients suffering from phenylketonuria, tyrosinemia, maple syrup urinary disease, hyperornithinemia, and citrullinemia are studied.

Amino Acids↗
PubMed2001

Ion fragmentation in an electrospray ionization mass spectrometer interface with different gases.

Nitrogen, argon, and krypton are used as curtain gases in an electrospray ionization mass spectrometer in an attempt to study the effect of these gases on the extent of ion fragmentation between the orifice and the skimmer of the interface region. A previously published collision model predicts that the degree of ion fragmentation increases with increasing mass of the curtain gas. However, the fragmentation yields are found to be the opposite to that expected. It is believed that the reversed trend with argon and krypton is caused by condensation of the gases within the free jet expansion between the orifice and the skimmer. A condensation parameter can be used to predict the degree of clustering of gases within a free jet expansion. When the condensation parameter is minimized, the predicted trend of fragmentation with mass is observed.

Argon↗
PubMed2001

Collision-induced dissociation of bradykinin ions in the interface region of an ESI-MS.

By applying different electric field strengths to the orifice-skimmer region of an electrospray ionization mass spectrometer, the rate of dissociation can be varied based on the amount of internal energy acquired by an ion through collisions with the curtain gas molecules. Both the Arrhenius equation and Rice-Ramsperger-Kassel (RRK) theory can be used to predict the rate of dissociation of internally excited molecules. A previously determined model for collision-induced dissociation is tested by comparison of predicted and experimentally observed orifice-skimmer potential differences for dissociation of ions. The rate of collision-induced dissociation of bradykinin ions is determined by monitoring the fragments produced in a mass spectrometer. The semi-quantitative model is found to yield effective predictions when accurate Arrhenius and RRK parameters are utilized.

Algorithms↗
PubMed2001

Emergent patterns of feedback strategies in performing a closed motor skill.

This sturdy identified self-chosen patterns of feedback usage and how they influenced accuracy and consistency in performing a closed motor skill. Learners completed 100 practice trials with 25 opportunities to receive knowledge of results (KR), the timing of which was self-chosen, followed by 25 no KR retention trials. The study identified two patterns of self-chosen KR, namely, a faded frequency pattern and a reversed faded frequency pattern. Analyses of variance indicated a significant trial block effect for accuracy in acquisition and no significant main effects or interactions associated with self-chosen patterns of KR. Quantification of regularity of KR patterns yielded correlations indicating that less regularity was associated with greater accuracy in performing the task. These results were congruent with previous research on self chosen feedback.

Adult↗
PubMed2000

Velocity-difference induced focusing of nucleotides in capillary electrophoresis with a dynamic pH junction.

Velocity-difference induced focusing (V-DIF) of nucleotides was achieved by using a dynamic pH junction in capillary electrophoresis (CE) with UV detection. The influence of specific analyte properties, such as nucleotide base structure, sugar structure, and degree of phosphorylation, is examined. The pKa values and borate complexation with vicinal diols are important factors that caused the focusing. Therefore, the pH and borate content in the sample and background electrolyte can be adjusted to optimize the focusing effect. This method allows the injection of large volumes of sample (approximately 300 nL), resulting in at least 50-fold improvement in concentration sensitivity. The detection limit of 4.0 x 10(-8) M for nucleotides can be achieved in favorable conditions. V-DIF can be also applied to nucleotide pool analysis from cell extracts to improve the concentration sensitivity of CE and to reduce the time-consuming steps of desalting and off-line preconcentration that are often required for assays of nucleotides from biological samples.

Animals↗
PubMed2000

Selective focusing of catecholamines and weakly acidic compounds by capillary electrophoresis using a dynamic pH junction.

A systematic study of selective analyte focusing in a multisection electrolyte system by capillary electrophoresis (CE) is presented. It was found that a dynamic pH junction between sample and background electrolyte zones can be used to focus zwitterionic catecholamines and weakly acidic compounds without the use of special ampholytes. Differences in pH and concentration of complexing agents, such as borate, in the sample and background electrolyte zones were determined to cause focusing through changes in the local velocity of the analyte in two different segments of the capillary. Velocity-difference induced focusing (V-DIF) of analytes using a dynamic pH junction allowed the injection of large sample volumes and significantly improved the concentration sensitivity of CE. Under optimized conditions, the limit of detection for epinephrine was determined to be about 4 x 10(-8) M (the original sample) with conventional UV absorbance detection. Moreover, separation efficiencies greater than a million theoretical plates can be achieved by focusing such large sample volumes into narrow zones. Multisection electrolyte systems, which lead to the formation of a dynamic pH junction, can be tuned toward improving the concentration sensitivity of specific analytes if their chemical properties are known.

Acids↗
PubMed1999

Analysis of epinephrine from fifteen different dental anesthetic formulations by capillary electrophoresis.

A robust method for the quantification of epinephrine from 15 different commercial dental anesthetic formulations is developed using CE. This work presents an extension to a method reported earlier. The solvability of several anesthetic compounds was improved through appropriate dilutions and the addition of sodium dodecyl sulfate to the separation background electrolyte. By controlling the mobility of the analyte at different pH values, a dilute solution of epinephrine is focused into a sharp zone with the injection of about 150 nl of anesthetic solution into the capillary. This on-column concentration technique extended the concentration detection limit of epinephrine to about 5.0 x 10(-7) M using a commercially available UV detector. A correlation plot between the measured and listed epinephrine concentration for the 15 dental anesthetic solutions demonstrated excellent accuracy of this method.

Anesthesia, Dental↗
PubMed1999

Analysis of gamma-carboxyglutamic acid content of protein, urine, and plasma by capillary electrophoresis and laser-induced fluorescence.

When the properties of an analyte are known, the separation system can be designed to make the analyte of interest migrate at either a much faster or a much slower velocity compared to other molecules in the sample matrix. A simple and sensitive method to analyze the gamma-carboxyglutamic acid (Gla) content of protein, urine, and plasma was developed using capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). The separation method is designed according to the specific properties of three amino acids of interest. The number of Gla residues from three vitamin K-dependent proteins were estimated by quantifying the amount of fluorescein thiocarbamyl derivative of Gla after alkaline hydrolysis and fluorescein isothiocyanate labeling. Human prothrombin, blood coagulation factor X, and bovine osteocalcin were calculated to have 10.0 +/- 0.7, 11.0 +/- 0.6, and 2.1 +/- 0.1 Gla residues per mole of protein, respectively, which agreed well with amino acid sequencing data. The analysis of free Gla content in urine and plasma was also demonstrated by this method. It was demonstrated that submicrograms of protein can be characterized by CE-LIF.

1-Carboxyglutamic Acid↗
PubMed1999

Undercarboxylation of recombinant prothrombin revealed by analysis of gamma-carboxyglutamic acid using capillary electrophoresis and laser-induced fluorescence.

The gamma-carboxyglutamic acid (Gla) content of several variants of human prothrombin has been measured by using capillary electrophoresis and laser-induced fluorescence (CE-LIF). Both plasma-derived prothrombin and recombinant prothrombin contain ten residues of Gla per molecule of protein. In contrast, a variant of human prothrombin (containing the second kringle domain of bovine prothrombin) was separated into two populations that differed in their Gla content. Direct measurement of the Gla content showed an association with the presence or absence of the calcium-dependent conformational change that is required for prothombinase function. Thus, the CE-LIF assay is useful in determining the carboxylation status of recombinant proteins.

1-Carboxyglutamic Acid↗
PubMed1998

Dynamic complexation of solutes in capillary electrophoresis.

The analyte migration behavior in any chemical separation system can be described using a single equation that unifies all areas of separation science. This equation can be used in capillary electrophoresis (CE) to design separation systems, and to study interactions between analytes and additives. By using individual capacity factors for each analyte species present in the system, and with the knowledge of the characteristics of each interaction, one can predict the analyte migration behavior in complicated CE systems, including systems with multiple 1:1 interactions and/or higher order interactions.

Binding Sites↗
PubMed1998

The effects of a mixture of charged and neutral additives on analyte migration behavior in capillary electrophoresis.

Multicomponent additives, such as derivatized cyclodextrins with various degrees of substitution, can be considered single-component additives as long as the fraction of each component remains constant. In this paper, equations are derived describing the effect of such additives on the migration behavior of analytes. These equations are used in the study of capillary electrophoresis (CE) systems with differentially charged cyclodextrins as additives. For weakly acidic analytes, the binding with highly negatively charged sulfobutyl ether beta-cyclodextrin (SBE-beta-CD) increases their negative electrophoretic mobility, while the binding with neutral hydroxypropyl-beta-cyclodextrin (HP-beta-CD) decreases their negative mobility. By obtaining the equilibrium constants and mobilities for each additive with each analyte (in this case, phenol, 2-naphthol and 1-naphthol), the migration behavior of these analytes in CE systems is quantitatively predicted at various concentrations of mixtures of the two additives. The properties of the contour lines in the binding isotherm surfaces of such CE systems are discussed.

2-Hydroxypropyl-beta-cyclodextrin↗
PubMed1998

The effect of complexation additives on analyte migration behavior in capillary electrochromatography.

In capillary electrochromatography (CEC), analytes often have different mobilities in the mobile phase, and often are involved in multiple equilibria. In this paper, the migration behavior of an analyte in CEC is described by a general equation in which individual capacity factors are used to describe the tendency of the analyte to exist as the various analyte species present in a separation system, and the effects of both field and equilibrium are accounted for. The resolution of two analytes is shown to be related linearly to the ratio of their migration rates. The effect of the electroosmotic flow (EOF) in CEC is more complicated than in CE because it is experienced only by a fraction of the analyte, whereas in CE, it is experienced by all analyte species. A procedure for calculating the electrophoretic mobility of the analyte based on the fraction of the analyte in the buffer is demonstrated. The effect of the EOF on resolution is also discussed.

Chromatography↗
PubMed1998

Recent developments towards a unified theory for separation science (minireview).

Various techniques for chemical separation can be described using a generally applicable theory. There are several schools of thought on how a unified separation science should be developed. The theories described include the mass balance equation (i.e. moving boundary), virtual migration distances, and the use of individual capacity factors.

Chromatography↗
PubMed1998

Quantitative assay for epinephrine in dental anesthetic solutions by capillary electrophoresis.

A simple and robust method for the separation and quantification of epinephrine in dental anesthetic solutions was developed. The method allows the direct injection of high salt solutions without sample pre-treatment. Large sample plugs (5.7% of the total capillary length) are used for epinephrine determination by selective analyte focusing in capillary electrophoresis. The concentration detection limit for epinephrine is about 5.0 x 10(-7) M (90 ng ml-1) with a commercial UV detector. The separation protocol was validated in terms of its precision, linearity, accuracy and specificity.

Anesthesia, Dental↗
PubMed1997

Quantitative description of migration behavior of porphyrins based on the dynamic complexation model in a nonaqueous capillary electrophoresis system.

The effect of an additive (Brij 35) on the mobilities of a group of porphyrin acids is quantitatively characterized based on a 1:1 dynamic complexation model. Varying additive concentration shifts the equilibrium and changes the viscosity of the background electrolyte. The equilibrium constant, the electrophoretic mobility of the free analyte, and the electrophoretic mobility of the complex are identified as the parameters necessary to describe the analytes' migration behavior. Several statistical methods for obtaining these parameters are discussed. The equilibrium constants and complex mobilities are calculated using three different linear regression methods. The weighted y-reciprocal method was preferred because it gives smaller error, and the data points are evenly distributed along the concentration axis. These values are confirmed using a nonlinear regression to ensure that the proper weighting was used in the linear regression plots. The parameters are then used to predict the apparent mobilities of the analytes over the entire additive concentration range, allowing the optimum separation conditions to be identified. For disc-like molecules, such as porphyrins, the mobility is determined by the orientation of the molecule in an electric field, in addition to their size and charge. The strength of binding between the porphyrins and Brij 35 depends on the number of binding sites and the solvation shell.

Buffers↗
PubMed1997

Quantitative description of analyte migration behavior based on dynamic complexation in capillary electrophoresis with one or more additives.

A comprehensive theory is proposed to describe the migration behavior of analytes in capillary electrophoresis (CE) when one or more additives are present in the buffer solution. This theory amalgamates and extends the previous work done by others. The capacity factor (k') in this theory is defined as the product of the equilibrium constant and the additive concentration, thus, k' changes linearly with additive concentration. The net electrophoretic mobility of an analyte is a function of k', therefore, it can be changed by varying the additive concentration. Three parameters are needed to predict the mobility of an analyte in a one-additive CE system: the mobility of the free analyte, the mobility of the complex, and the equilibrium constant for the analyte-additive interaction (which determines the fraction of the free analyte at different additive concentrations). When additives are used, the change in viscosity obscures this relationship, therefore, a viscosity correction factor is required to convert all mobilities to an ideal state where the viscosity remains constant. The migration behavior of an analyte in a solution with multiple additives can be predicted and controlled, once the equilibrium constants of the interactions between the analyte and each of the additives are obtained separately. beta-Cyclodextrin and hydroxypropyl-beta-cyclodextrin are used as additives and the migration behavior of phenol, p-nitrophenol, and benzoic acid are studied as a model system to verify this theory. When the necessary viscosity correction factor is included, the net electrophoretic mobilities of the analytes obtained from experimental results agree with the values predicted by the theory based on dynamic complexation. Although only experiments with one and two additives were carried out to verify the theory, the equations apply to situations when more than two additives are used. The relationship between the theories of electrophoresis and chromatography is clarified.

2-Hydroxypropyl-beta-cyclodextrin↗
PubMed1997

Redefining the separation factor: a potential pathway to a unified separation science.

Understanding the separation process in capillary electrophoresis (CE) leads to the unification of the theories for separation science. While the separation of analytes is governed by equilibria in chromatography, and by (centrifugal) field in ultracentrifugation, the separation in CE is governed by both equilibria and (electric) field. Therefore, a comprehensive separation theory that describes the separation process of analytes in CE should be able to describe the separation processes in both chromatography and ultracentrifugation. In this paper, we propose that individual capacity factors for each analyte species be used to describe the migration behavior of an analyte. The effect of field on each analyte species, as well as the effect of equilibria are considered in deriving a generalized equation that is applicable for all separation techniques. The separation factor defined at present does not directly relate to the migration rates of the analytes, and therefore can not be used in a generalized theory. We propose that the ratio of the migration rates of a pair of analytes (gamma) should be used as the separation factor, instead of the ratio of the two capacity factors. When gamma is used to describe the separation of two closely migrating analytes, all separation techniques have the same resolution equation.

Chromatography↗