Separation method based on affinity reaction between magnetic and nonmagnetic particles for the analysis of particles and biomolecules.

A separation method is reported for particle and biochemical analysis based on affinity interactions between particle surfaces under magnetic field. In this method, magnetic particles with immunoglobulin G (IgG) or streptavidin on the surface are flowed through a separation channel to form a deposition matrix for selectively capturing nonmagnetic analytes with protein A or biotin on the surface due to specific antigen (Ag)--antibody (Ab) interactions. This separation method was demonstrated using model reactions of IgG--protein A and streptavidin-biotin on particle surface. The features of this new separation method are (1) the deposited Ag-Ab complex can be examined and further analyzed under the microscope, (2) a kinetic study of complex binding is possible, and (3) the predeposited matrix can be formed selectively and changed easily. The detection limits were about 10(-11) g. The running time was less than 10 min. The selectivities of studied particles were 94% higher than those of label-controlled particles. This method extends the applications of analytical magnetapheresis to nonmagnetic particles. Preliminary study shows that this separation method has a great potential to provide a simple, fast, and selective analysis for particles, blood cells, and immunoassay related applications.

Determination of magnetic susceptibility of various ion-labeled red blood cells by means of analytical magnetapheresis.

Analytical magnetapheresis is a newly developed technique for separating magnetically susceptible particles. The magnetically susceptible particles are deposited on a bottom plate after flowing through a thin (< 0.05 cm) separation channel under a magnetic field applied perpendicular to the flow. Particles with various magnetic susceptibilities can be selectively deposited and separated by adjusting the applying magnetic force and flow rates. Magnetic susceptibility is an important parameter for magnetic separation. Magnetic susceptibility determination of various ion-labeled red blood cells (RBCs) using analytical magnetapheresis with a simple theoretical treatment is reported in this study. Susceptibility determination is based on the balance between maximal channel flow rate and magnetically induced flow rate for deposition. We tried a new approach to determine particle magnetic susceptibilities using a balance of magnetic and drag forces to control magnetically induced particle velocities. The Er3+, Fe3+, Cu2+, Mn2+, Co2+, and Ni2+ ions were used to label RBC at various labeling concentrations for susceptibility determination. The susceptibilities determined for various ion-labeled RBC under two magnetic field intensities fell within a 10% range. The average viabilities of various ion-labeled RBCs were 96.1 +/- 0.8%. The susceptibility determination generally took less than 10 min. Determined susceptibilities from analytical magnetapheresis differed by 10% from reference measurements using a superconducting quantum interference device (SQUID) magnetometer. The cost and time for analysis is much less using analytical magnetapheresis. This technique can provide a simple, fast, and economical way for particle susceptibility determinations.

Solid-phase microextraction coupled with liquid chromatography for determination of beta-carotene in food.

Beta-carotene in vegetables and nutritional products is analyzed using solid-phase microextraction (SPME) coupled with liquid chromatography (LC) to improve the speed of analysis and to reduce the consumption of organic solvents. The relative standard deviations (RSDs) of this analytical method for beta-carotene determinations in vegetables and nutritional products are approximately 10% and 5%, respectively. The amount of beta-carotene was found to vary from 0.35 +/- 0.05 ppm to 76.5 +/- 6.9 ppm for several vegetables in Taiwan. This method was linear over the range of 0.4-40 ppm with correlation coefficients higher than 0.997. The experimentally determined level of beta-carotene in nutritional products varied from 3.8 +/- 0.2 ppm to 24.6 +/- 1.1 ppm following SPME-LC. The recoveries of beta-carotene for these measurements following SPME were all higher than 97% +/- 2% (n = 3). The detection limits of beta-carotene for this method were from 0.027 to 0.054 ppm. Conventional solvent extractions take approximately 4-6 h for extraction and reconcentration but SPME takes approximately 1 h. From several tens to hundreds of milliliters, organic solvents can be saved using SPME. SPME provides better analyses on beta-carotene than conventional solvent extraction for nutritional products in terms of speed, precision, simplicity, and solvent consumption.

Rapid diffusion coefficient measurements using analytical SPLITT fractionation: application to proteins.

This work reports a new technique for the rapid measurement of diffusion coefficients using a special flow cell called a SPLITT cell. Such SPLITT cells, designed for continuous SPLITT fractionation, utilize differential transport across a thin (approximately 100 microns) ribbonlike lamina flowing through the cell. The liquid stream in the cell is split into two substreams at the outlet. The relative concentration in the two outlet substreams of a dilute component introduced into one of two inlet substreams is mathematically related to the appropriate transport coefficient, in this case the diffusion coefficient D. Thus D can be calculated from measured values of relative outlet concentrations. This approach has been tested using a number of dilute protein solutions. The results at different stream-splitting ratios are reasonably self-consistent and agree with most literature values for D within 5%. This approach provides a simple, rapid, and predictable means for determining the diffusion coefficients of proteins and other substances.