Apparatus and method of multi-dimensional chemical separation

A method of two-stage thermal modulation for generating concentration pulses in a fluid stream flowing through a tube, said tube comprising an inlet, an outlet, a first portion which is a length of said tube comprising a first stage, and a final portion which is a length of said tube comprising a outlet stage, said method comprising the steps of:

• (a) creating a flow of carrier fluid in a direction through said tube to produce a carrier fluid flow;
• (b) introducing a sample into the carrier fluid flow, said sample comprising one or more sample substances;
• (c) manipulating the temperature of the first stage to cause the sample to be retained;
• (d) manipulating the temperature of the outlet stage such that the sample is retained;
• (e) accumulating within the first stage for a period of time sample substances carried thereinto by the carrier fluid, thus forming a first concentration;
• (f) manipulating the temperature of the first stage to release the first concentration into the carrier fluid flow in the form of a first concentration pulse;
• (g) carrying the first concentration pulse in said carrier fluid flow toward said outlet stage;
• (h) accumulating said first concentration pulse at an inlet of said outlet stage so as to focus and hold therein for a period of time, sample substances of said first concentration pulse carried by the carrier fluid, thus forming an outlet concentration more compact in distance than said first concentration pulse;
• (i) positioning the temperature of the first stage so as to resume accumulating therein for a period of time sample substances carried thereinto by the carrier fluid;
• (j) positioning the temperature of the outlet stage so as to release said outlet concentration into the carrier fluid flow in the form of an outlet concentration pulse, said outlet concentration pulse being of shorter duration than said first concentration pulse;
• and
• (k) positioning the temperature of the outlet stage such that the sample is retained.

A method of chemical separation comprising one or more portion of tube forming one or more capillary electrophoretic column, said method further comprising the steps of:

• (a) injecting a sample composed of two or more substances into a first dimension of electrophoresis;
• (b) carrying said sample along said first dimension of electrophoresis in a direction so as to cause a first separation of said sample into a first series of bands dispersed during a first separation time period, each band of said first series of bands being composed of one or more substance and being either separated from or partially overlapping with one or more other band in said first series of bands;
• (c) accumulating a portion of the sample in a modulator positioned between said first dimension of electrophoresis and a final dimension of electrophoresis disposed in serial relationship with said first dimension of electrophoresis;
• (d) transmitting said portion of the sample from said modulator as a concentration pulse into said final dimension of electrophoresis;
• (e) carrying said portion of the sample along said final dimension of electrophoresis in a direction so as to cause a final separation of said portion of the sample into a final series of bands dispersed during a final separation time period;
• and
• (f) repeating steps (c), (d), and (e) during said first separation time period so as to generate a multiplicity of said final separations and a multiplicity of said final series of bands during a multiplicity of said final separation time periods such that,
• said first series of bands emerging from said first dimension of electrophoresis is submitted to and transmitted through said final dimension of electrophoresis with fidelity, and
• an electrophoretic parameter affecting retention of said final dimension of electrophoresis is varied as a function of the progress of said first dimension of electrophoresis such that, said final series of bands emerges from said final dimension of electrophoresis within a limited and defined range of retention times and said comprehensive two-dimensional electrophorogram is substantially orthogonal.

A method of two-dimensional chromatographic separation comprising the steps of:

• (a) injecting a sample composed of various sample substances into a first dimension of chromatographic separation;
• (b) carrying said sample along said first dimension of chromatographic separation so as to cause a first resolution of said sample into a series of bands;
• (c) accumulating a portion of the sample between said first dimension of chromatographic separation and a second dimension of chromatographic separation disposed in serial relationship with said first dimension of chromatographic separation;
• (d) transmitting the accumulated portion of sample as a sharp wave front to an inlet of the second dimension of chromatographic separation;
• (e) carrying the portion of sample along the second dimension of chromatographic separation so as to cause a further resolution of said portion of sample;
• repeating steps (c), (d), and (e) so as to generate a multiplicity of said sharp wave fronts and a multiplicity of said further resolutions by said second dimension of chromatographic separation, without materially changing said first resolution;
• and
• detecting said multiplicity of further resolutions so as to generate a multiplicity of second dimension chromatograms together comprising a two-dimensional chromatogram of said sample.