Metal speciation using capillary electrophoresis--inductively coupled plasma atomic emission spectrometry and polytetrafluoroethylene capillaries

A capillary electrophoresis--inductively coupled plasma atomic-emission spectrometry (CE-ICP-AES) system using a polytetrafluoroethylene (PTFE) capillary has been developed. The CE-ICP interface was a modified concentric nebulizer. The PTFE capillary (50 microm internal diameter) was used as the central capillary of the nebulizer. Using the PTFE capillaries, the solution flow rate induced by the carrier gas flow was smaller than that of glass capillary. Solution flow was mainly induced by the CE electric field. Baseline separation of Ba2+/Mg2+ ion pair using simple buffer solution of 0.014 M sodium acetate was reported. Separation and correlation of metal species in metallothioneins (MT-1 and MT-2 in MT) of rabbit liver using the CE-ICP system were also discussed.     

Interface for capillary electrophoresis coupled with inductively coupled plasma atomic emission spectrometry.

A modified concentric nebulizer was used as the interface to couple capillary electrophoresis (CE) to inductively coupled plasma atomic emission spectrometry (ICP-AES). The CE capillary replaces the central tube of the concentric nebulizer. The tip of the nebulizer tapers slowly to allow uncertainty in the position of the capillary. A platinum wire was inserted into the CE capillary to provide electrical connection to the CE power supply. pH changes inside the capillary due to electrolysis of the background buffer electrolyte was small and has minimal effects on the CE separation. The peak broadening effects due to the nebulizing gas flow, however, were significant. Resolution decreases quickly when the flow-rate of the carrier gas increases. Sample stacking technique was used to improve the resolution of species of opposite charge, e.g., Cr(VI) vs. Cr(III) ions. Detection limit of Cr based on peak area is approximately 10 ppb for the CE-ICP-AES system.     

Identification of a metallothionein in Synechococcus by capillary electrophoresis hyphenated with inductively coupled plasma mass spectrometry.

A home-made system hyphenating capillary electrophoresis with an inductively coupled plasma mass spectrometer (CE-ICP-MS) for cadmium speciation of protein-binding and free cadmium ions in solution is presented. The CE-ICP-MS interface consisted of an acrylic block with an internal volume ca. 20 microL in which a platinum electrode, a capillary column, and a connection to an ICP nebulizer were inserted. A make-up electrolyte solution containing 50 mmol L(-1) Tris-HCl buffer solution (pH 9.0) was continuously flowed through the interface to the ICP nebulizer. The separation of free Cd ions, Cd-cysteine, and Cd bounded to metallothionein (MT) isoforms from rabbit liver was carried out by capillary electrophoresis, and the analytes were detected by ICP-MS. The feasibility to isolate metallothionein compounds extracted from the cyanobacterium Synechococcus PCC7942 was demonstrated. The Cd binding proteins were induced in Synechococcus PCC7942 and further analyzed by CE ICP-MS.     

A new interface for combining capillary electrophoresis with inductively coupled plasma-mass spectrometry

This paper describes the development and design of a new, efficient, simple and robust interface for coupling capillary electrophoresis (CE) with inductively coupled plasma-mass spectrometry. The interface is based on a modified microconcentric nebulizer which permits a low flow rate of about 6 μL/min in the free aspiration mode. This interface construction provides an electrical connection for stable electrophoretic separations and adapts the flow rate of the electro-osmotic flow inside the CE capillary to the flow rate of the nebulizer for efficient transport of the analytes into the plasma. By optimization of the fluid mechanical properties the interface prevents the nebulizer from causing any laminar flow in the CE capillary and thus the high resolution power of CE can be preserved. Furthermore, this new device permits independent optimization of the nebulization from the CE whereby exact positioning of the CE capillary is not necessary, thus enabling fast exchange. A low dead volume spraychamber has been constructed which circumvents any band broadening of the sharp CE signals. Peak widths down to 3.5 s comparable to CE with UV detection are possible. Received: 5 February 1999 / Revised: 21 April 1999 / Accepted: 23 April 1999     

A new interface for combining capillary electrophoresis with inductively coupled plasma-mass spectrometry

This paper describes the development and design of a new, efficient, simple and robust interface for coupling capillary electrophoresis (CE) with inductively coupled plasma-mass spectrometry. The interface is based on a modified microconcentric nebulizer which permits a low flow rate of about 6 μL/min in the free aspiration mode. This interface construction provides an electrical connection for stable electrophoretic separations and adapts the flow rate of the electro-osmotic flow inside the CE capillary to the flow rate of the nebulizer for efficient transport of the analytes into the plasma. By optimization of the fluid mechanical properties the interface prevents the nebulizer from causing any laminar flow in the CE capillary and thus the high resolution power of CE can be preserved. Furthermore, this new device permits independent optimization of the nebulization from the CE whereby exact positioning of the CE capillary is not necessary, thus enabling fast exchange. A low dead volume spraychamber has been constructed which circumvents any band broadening of the sharp CE signals. Peak widths down to 3.5 s comparable to CE with UV detection are possible. Received: 5 February 1999 / Revised: 21 April 1999 / Accepted: 23 April 1999     

In-column fiber-optic laser-induced fluorescence detection for CE

A highly sensitive in-column fiber-optic LIF detector for CE has been constructed and evaluated. In this detection system, a 457-nm diode-pumped solid-state blue laser was used as the excitation light source and an optical fiber (40 mum od) was used to transmit the excitation light. One end of the optical fiber was inserted into the separation capillary and was in situ positioned at the detection window. The other end of the fiber was protruded from the capillary to capture the excitation light beam from the blue laser. Fluorescence emission was collected by a 40 x microscope objective, focused on a spatial filter, and passed through a yellow color filter before reaching the photomultiplier tube. The present CE-fluorescence detection is a simple and compact optical system. It reduces the laser scattering effect from the capillary and fiber as compared to the conventional LIF detection for CE. Its utility was successfully demonstrated by the separation and determination of D-penicillamine labeled with naphthalene-2,3-dicarboxaldehyde. The detection limit was 0.8 nM (S/N = 3). The present detection scheme has been proven to be attractive for sensitive fluorescence detection for CE.     

一种新型毛细管电泳—电感耦合等离子体光谱法接口装置

毛细管电泳是一种高效、简便的分离方法，已被用于生物、环境及临床等试样的分离及分析[1]．检测技术在毛细管电泳中占有重要的地位，目前，在柱紫外可见及荧光检测是两种广为接受的检测方法，但其检测灵敏度仅为10－5～10－6mol／L[2]．电感耦合等离子体光谱（ICP－AES／MS）是一种灵敏的元素选择性的分析方法，已被广泛地用于各种试样中元素分析．近年来，该方法作为色谱及毛细管电泳的检测器，被用于元素的形态分析[3]．在毛细管电泳（CE）与ICP光谱连用技术中，挑战性的工作是设计一种能把CE与ICP相连的接口．目前已有几种接口…     

A novel high-temperature (360 degrees C)/high-pressure (30 MPa) flow system for online sample digestion applied to ICP spectrometry

A flow injection sample digestion system has been developed comprising an indirectly electrically heated Pt/Ir capillary. Such a capillary allows reaction temperatures of up to 360 degrees C and pressures of up to 30 MPa (300 bar) and withstands concentrated acids. This temperature is 130 degrees C to 160 degrees C higher compared to the operating temperatures of microwave heated flow systems. A combination of an ultrasonic nebulizer and membrane desolvator serves as an interface between the flow digestion system and an ICP/AES spectrometer. The membrane desolvator removes interfering gaseous digestion products so effectively before the sample stream enters the plasma that the measured residual carbon concentration falls in the region of the detection limit of ICP/OES measurements. Sewage sludge samples were digested using nitric acid and the elemental traces online determined. The detection limits related to the original dry substances amount to the lower microg/g range.     

Capillary electrophoresis-inductively coupled plasma-mass spectrometry: a report on technical principles and problem solutions, potential, and limitations of this technology as well as on examples of application

This paper summarizes some basic principles of capillary electrophoresis (CE), inductively coupled plasma-mass spectrometry (ICP-MS), and coupling of both devices. Especially the interfacing is described in detail. A special focus is drawn to various interface developments reported in literature and technical problems, i.e., requirements to the interface setup and respective solutions. Nowadays, typically sheath flow-based interfaces are used. The sheath flow fulfills two requirements of hyphenation, (i) the closing of the electrical circuit of CE and (ii) the feeding of the used nebulizer with an adequate flow rate. In the beginning of CE-ICP-MS coupling predominantly home-made interface-nebulizer constructions were developed and tested for various speciation problems. Now increasingly such laboratory-constructed interfaces are left. Mostly commercial nebulizers are employed being combined with commercially available tee or cross fittings to connect the CE capillary to the electrode, the additional sheath flow, and the nebulizer. Due to the low sample amounts and low flow rates from CE, such nebulizers are typically low-flow nebulizers like, e.g., the microconcentric nebulizer (MCN) and the direct injection nebulizer (DIN). However, there are also reports on couplings using standard Meinhard systems. Still the control and reduction of a siphoning sucting flow and sufficient detection limits are the major problems in hyphenating CE to ICP-MS. Different solutions are reported on these problems and summarized here. Finally numerous applications are reported. Mostly, applications are performed on speciation of selenium, arsenic, metallothionein isoforms, mercury, or cobalt.     

A simple microfluidic system for efficient capillary electrophoretic separation and sensitive fluorimetric detection of DNA fragments using light-emitting diode and liquid-core waveguide techniques

A miniaturized CE system has been developed for fast DNA separations with sensitive fluorimetric detection using a rectangle type light-emitting diode (LED). High sensitivity was achieved by combining liquid-core waveguide (LCW) and lock-in amplification techniques. A Teflon AF-coated silica capillary on a compact 6x3 cm baseplate served as both the separation channel for CE separation and as an LCW for light transmission of fluorescence emission to the detector. An electronically modulated LED illuminated transversely through a 0.2 mm aperture, the detection point on the LCW capillary without focusing, and fluorescence light was transmitted to the capillary outlet. To simplify the optics and enhance collection of light from the capillary outlet, an outlet reservoir was designed, with a light transmission window, positioned directly in front of a photomultiplier tube (PMT), separated only by a high pass filter. Automated sample introduction was achieved using a sequential injection system through a split-flow interface that allowed effective release of gas bubbles. In the separation of a phiX174 HaeIII DNA digest sample, using ethidium bromide as labeling dye, all 11 fragments of the sample were effectively resolved in 400 s, with an S/N ratio comparable to that of a CE system with more sophisticated LIF.     

A novel high-temperature (360 °C)/high-pressure (30 MPa) flow system for online sample digestion applied to ICP spectrometry

A flow injection sample digestion system has been developed comprising an indirectly electrically heated Pt/Ir capillary. Such a capillary allows reaction temperatures of up to 360?°C and pressures of up to 30 MPa (300 bar) and withstands concentrated acids. This temperature is 130?°C to 160?°C higher compared to the operating temperatures of microwave heated flow systems. A combination of an ultrasonic nebulizer and membrane desolvator serves as an interface between the flow digestion system and an ICP/AES spectrometer. The membrane desolvator removes interfering gaseous digestion products so effectively before the sample stream enters the plasma that the measured residual carbon concentration falls in the region of the detection limit of ICP/OES measurements. Sewage sludge samples were digested using nitric acid and the elemental traces online determined. The detection limits related to the original dry substances amount to the lower μg/g range.     

Design and performance of a sheathless capillary electrophoresis/mass spectrometry interface by combining fused-silica capillaries with gold-coated nanoelectrospray tips

A simple sheathless capillary electrophoresis (CE)/mass spectrometry (MS) interface was constructed by combining widely used nanospray needles with fused-silica capillaries and it was successfully applied for the separation of peptides. The end of the CE capillary was pulled to a taper, etched and then fitted into the metal-coated nanospray borosilicate capillary. The nanospray needle can be used for several CE runs, but it can be easily and rapidly changed in the case of accidental breakage or evaporation of the coating. A fast capillary electrochromatographic method was also developed for MS analysis of peptides containing numerous basic amino acids.     

Capillary electrophoresis-inductively coupled plasma mass spectrometry interface with minimised dead volume for high separation efficiency

A modified microconcentric nebulizer (MCN-100) has been used to construct an improved interface with minimised liquid and gas phase dead volumes for the coupling of capillary electrophoresis (CE) and inductively coupled plasma mass spectrometry (ICP-MS). A plate number of 3.6 x 10(6) plates m(-1) has been achieved. This is an order of magnitude better than the results previously reported for CE-ICP-MS. The separation efficiency of the system is demonstrated by the baseline separation of eight rare earth metals within a time span of 14.6 s. The system was used to control the purity of vitamin B12 and glutaredoxin 2 from Escherichia coli.     

Use of a parallel path nebulizer for capillary-based microseparation techniques coupled with an inductively coupled plasma mass spectrometer for speciation measurements

A low flow, parallel path Mira Mist CE nebulizer designed for capillary electrophoresis (CE) was evaluated as a function of make-up solution flow rate, composition, and concentration, as well as the nebulizer gas flow rate. This research was conducted in support of a project related to the separation and quantification of cobalamin (vitamin B-12) species using microseparation techniques combined with inductively coupled plasma mass spectrometry (ICP-MS) detection. As such, Co signals were monitored during the nebulizer characterization process. Transient effects in the ICP were studied to evaluate the suitability of using gradients for microseparations and the benefit of using methanol for the make-up solution was demonstrated. Co signal response changed significantly as a function of changing methanol concentrations of the make-up solution and maximum signal enhancement was seen at 20% methanol with a 15 μl/min flow rate. Evaluation of the effect of changing the nebulizer gas flow rates showed that argon flows from 0.8 to 1.2 l/min were equally effective. The Mira Mist CE parallel path nebulizer was then evaluated for interfacing capillary microseparation techniques including capillary electrophoresis (CE) and micro high performance liquid chromatography (μHPLC) to inductively coupled plasma mass spectrometry (ICP-MS). A mixture of four cobalamin species standards (cyanocobalamin, hydroxocobalamin, methylcobalamin, and 5′ deoxyadenosylcobalamin) and the corrinoid analogue cobinamide dicyanide were successfully separated using both CE-ICP-MS and μHPLC-ICP-MS using the parallel path nebulizer with a make-up solution containing 20% methanol with a flow rate of 15 μl/min.     

A new interface used to couple capillary electrophoresis with an inductively coupled plasma mass spectrometry for speciation analysis

In this work, a novel and high-efficiency interface has been developed in coupling CE with inductively coupled plasma MS (ICPMS). The interface completely avoids laminar flow in CE capillary caused by the suction of nebulizer, and can be easily and stably operated at room temperature with high analyte transport efficiency to ICPMS. The new interface has a liquid dead volume smaller than 5 nL, which was much smaller than those (65-2500 microL) reported previously for other interfaces. All above features led to a higher sensitivity and a better electrophoretic resolution for CE-ICPMS coupled with this new interface. With the help of this new interface, we have successfully separated and determined five species of arsenic, As(III), As(V), monomethylarsonic acid, dimethylarsinic acid and p-aminophenylarsonic acid using CE-ICPMS within 11 min with a detection limit of 0.046-0.075 ng/mL and an RSD of 2-6% (n=6).     

The influence of the sample introduction system on signals of different tin compounds in inductively coupled plasma-based techniques

The influence of the sample introduction system on the signals obtained with different tin compounds in inductively coupled plasma (ICP) based techniques, i.e., ICP atomic emission spectrometry (ICP–AES) and ICP mass spectrometry (ICP–MS) has been studied. Signals for test solutions prepared from four different tin compounds (i.e., tin tetrachloride, monobutyltin, dibutyltin and di-tert-butyltin) in different solvents (methanol 0.8% (w/w), i-propanol 0.8% (w/w) and various acid matrices) have been measured by ICP–AES and ICP–MS. The results demonstrate a noticeable influence of the volatility of the tin compounds on their signals measured with both techniques. Thus, in agreement with the compound volatility, the highest signals are obtained for tin tetrachloride followed by di-tert-butyltin/monobutyltin and dibutyltin.The sample introduction system exerts an important effect on the amount of solution loading the plasma and, hence, on the relative signals afforded by the tin compounds in ICP–based techniques. Thus, when working with a pneumatic concentric nebulizer, the use of spray chambers affording high solvent transport efficiency to the plasma (such as cyclonic and single pass) or high spray chamber temperatures is recommended to minimize the influence of the tin chemical compound. Nevertheless, even when using the conventional pneumatic nebulizer coupled to the best spray chamber design (i.e., a single pass spray chamber), signals obtained for di-tert-butyltin/monobutyltin and dibutyltin are still around 10% and 30% lower than the corresponding signal for tin tetrachloride, respectively. When operating with a pneumatic microconcentric nebulizer coupled to a 50 °C-thermostated cinnabar spray chamber, all studied organotin compounds provided similar emission signals although about 60% lower than those obtained for tin tetrachloride. The use of an ultrasonic nebulizer coupled to a desolvation device provides the largest differences in the emission signals, among all tested systems.     

On-line conversion and determination of artemisinin using a flow-injection capillary electrophoresis system

A novel, rapid, and simple capillary electrophoresis (CE) method has been developed for the determination of antimalarial artemisinin by on-line treatment with alkaline. By on-line reaction, artemisinin was automatically and reproducibly converted to the strongly UV-absorbing compound, Q292, by treating it with 0.20 mol/L NaOH solution for 3 min at 40 degrees C. Analysis was carried out in less than 12 min after conversion of artemisinin in a flow injection (FI) system that was coupled to CE equipment via a split-flow interface cell, and a sampling frequency of 8 h(-1) is achievable. The on-line conversion method has been applied to the determination of artemisinin in the traditional Chinese herbal drug Artemisia annua L., and the results are satisfactory.     

Optical fiber light-emitting diode-induced fluorescence detection for capillary electrophoresis

A highly sensitive optical fiber light-emitting diode (LED)-induced fluorescence detector for CE has been constructed and evaluated. In this detector, a violet or blue LED was used as the excitation source and an optical fiber with 40 microm OD was used to transmit the excitation light. The upper end of the fiber was inserted into the separation capillary and was situated right at the detection window. Fluorescence emission was collected by a 40 x microscope objective, focused on a spatial filter, and passed through a cutoff filter before reaching the photomultiplier tube. Output signals were recorded and processed with a computer using in-house written software. The present CE/fluorescence detector deploys a simple and inexpensive optical system that requires only an LED as the light source. Its utility was successfully demonstrated by the separation and determination of amino acids (AAs) labeled with naphthalene-2,3-dicarboxaldehyde (NDA) and FITC. Low detection limits were obtained ranging from 17 to 23 nM for NDA-tagged AAs and 8 to 12 nM for FITC-labeled AAs (S/N=3). By virtue of such valuable features as low cost, convenience, and miniaturization, the presented detection scheme was proven to be attractive for sensitive fluorescence detection in CE.     

新型毛细管电泳化学发光系统应用于氨基酸分离测定

本研究建立一种以过氧草酸酯为化学发光体系的新型毛细管电泳化学发光系统,并将该系统应用于氨基酸的分离测定.该系统设计了新型的电极模式与反应混和模式以获得高的信号稳定性与检测灵敏度.以一端与高压电源负极相连的铂丝连接到不锈钢注射针头为接地电极,电泳毛细管穿过注射针头,且其末端到达注射针头的出口,电泳毛细宇航局末端与化学发光试剂在反应池中以相对的方向靠近.实验结果表明,该系统稳定性好、分离效率高、灵敏度高、精密度好,对天冬氨酸与亮氨酸的检出限分别为2.0和 1.1 nmol/L.峰高及迁移时间的相对标准偏差分别为2.3%～3.8%和1.2%～1.5%.     

On-line coupling of gas diffusion to capillary electrophoresis

On-line gas diffusion has been coupled to a capillary electrophoresis system (CE) via a specially designed interface. The sample is merged with a modifying solution, e.g., a strong acid, in a flow system to transform the analytes of interest into their respective gaseous forms. These transformed, gaseous analytes permeate through a PTFE membrane into an acceptor stream comprising of a tris-buffer. The continuously flowing acceptor stream is led into an injector forming an integrated part of a flow injection analysis (FIA) system. The sample receiving carrier stream in the FIA system, a chromate buffer, brings the sample, 50 mul, to the FIA-CE interface into which one end of a separation capillary has been inserted. A small portion of the injected sample enters the capillary (electrokinetic injection) and separation takes place. A UV detector is placed at the other capillary end and a run potential of 25 kV is applied to two platinum electrodes positioned in the flow system. Multiple sample injections can be performed in one uninterrupted electrophoretic run. A typical sampling frequency is 15 h(-1); each run may result in quantitation of at least five anions. The overall repeatability is in the range 1.8-3.6% (RSD). The technique has been applied to the analysis of real samples such as soft drinks, vinegar and wine. Selective discrimination of anions which are unable to form volatile species is accomplished. No off-line sample pre-treatment is needed.