Supercritical fluid chromatography laser wave mixing detection methods and apparatus

This invention relates to methods and apparatus of a combination of laser wave mixing technology with diagnostic flow technology with embodiments describing supercritical fluid chromatography. The mix of these technologies together with instant detection amounts still haven’t been seen in the field.


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Laser wave mixing has been described in many patents, journals and articles. Having greatest terms of embodiments of the invention described herein are Tong has described degenerate four wave-mixing and apparatus therein in U.S. Pat. Nos.5,600,444 and 6,141,094 and Patent Application 2006263777. These explain apparati and techniques that in their abilities are capable of analyzing small quantities of analytes down to a detection level of attomoles. They use different complements ofanalysis systems including HPLC and HCPE and a gas phase atomizer type spectroscopy. Furthermore, the dissertation”Protein Analysis at the Single Cell Level by Nonlinear Laser Wave-Mixing Spectroscopy for High Throughput Capillary ElectrophoresisApplications” from Sadri’s PhD dissertation N.C. State in 2008 relates similar apparati discussed in the Tong patents that reach the levels of discovery of yoctomoles (10.sup.-24). The termed articles, dissertations and patents are incorporated byreference in their entirety. These references provide a background into the concepts, alterations and variations upon the tech that are self explanatory. Similarly, supercritical fluid chromatography (SFC) has been clarified and explain many patentsand journal posts. A review article gives a fantastic illustration of the technology as used with chiral compounds”Enantioseparations in super- and subcritical fluid chromatography” Gerald Terfloth, J. Chromatogr. A 906 (2001) 301-307. This reference givesa modern perspective of how SFC can be used using the various column technologies and instrumentation techniques. Some advantages of SFC include: greater efficiency, wider range of selectivity, longer column life, lower solvent and waste disposal price, with lesstoxic solvents and faster run time and overall more environmentally cleaner than other techniques.

As used in this specification and in the appended claims, the singular forms “a,” an” and “the” include plural references unless the content clearly dictates otherwise.

In describing the invention and embodiments, the following terms will be used and are meant to be defined as indicated below. If any terms are not fully defined, then the normal usage as used in the art will fill some openings in theunderstanding of the terminology.

Laser: is a device that creates a beam of light where all the photons are in a coherent state–usually with the same frequency and phase. Among the additional effects, this usually means that the light from a laser can be tightly focused and can notdiverge considerably, causing the conventional laser beam. In free space, the beams inside and outside the cavity are often Gaussian distributed and therefore are highly collimated with very modest divergence. The space over which the laser beam remainscollimated depends on the square of the beam diameter whilst divergence angle varies slightly with the beam diameter.

Collimating: is the procedure of making light rays parallel by a mixture of diverging light beams or rays, and so will propagate slowly as it propagates. The term is related to”collinear” and implies light that does not disperse withdistance (ideally), or that will disperse minimally (in fact ). A perfectly collimated beam free of divergence can’t be created due to diffraction, but mild can be roughly collimated by numerous processes, for example by means of acollimator. A focusing lens using large focal point (1000 mm) is an illustration collimating lens.

Diagnostic flow technology: Is a good state technology by means of a series of pumps or pump such as mechanics (such as electroosmotic flow, electrophoretic flow, capillary action, siphoning, pressure, imploding gas bubbles and the like) and apparatimove analytes in a sample set area to an investigation area which contain of numerous detectors types such as photodiode arrays (PDA), ultraviolet-visible (UV-VIS) spectrometers, charge coupled device (CCD) (such as Fourier Transform Infrared(FT-IR)}, Nuclear Magnetic Resonance (NMR) detectors, Refractive Index spectrometers (RI), fluorescence detectors, radiation photomultipliers, and so on. Flow can be accomplished through fluids, fluids, gasoline or alternative means pumped or alternative means driventhrough a series of channels and mediums (such as tubing or silica gels) to move analytes from one stage to another. Examples would include but not restricted to Liquid Chromatography (LC) (which would further comprises variants such as micellar, ionexchange and the like), Reverse Phase High Performance Liquid Chromatography (RP-HPLC), Gas Chromatography (GC), High Performance Capillary Electrophoresis (HPCE), Capillary Zone Electrophoresis (CZE), Supercritical Fluid Chromatography (SFC),Sub-critical Fluid Chromatography (SubFC), Inductively Coupled Plasma (ICP), and so on. Every technology is unique unto its own with positives and negatives propagating from each in achieving the needs of the user. By way of example, capillaryelectrophoresis has ecological positives in utilizing very little hazardous materials but has adverse issues in what substances can be an analyzed and in what solvents are compatible.

Focal spot: a place or point onto which collimated light parallel to the axis of a lens is focused. This place of light could be expanded and contracted in different shapes and geometries by some means including a cylindrical lens.

Absorptive interaction: interaction of analytes in a flow cell chamber or multi channel chamber that makes a sign through the beam wave mixing when both input beams form light induced gratings and an input beam is diffracted away thisgrating when analytes absorb the excitation light beam. The excited molecules in the form of interference patterns discharge their heat energy to surrounding solvent or matrix molecules, producing dynamic thermal gratings, and consequently, refractive indexgratings. The incoming photons from the probe beam diffract off the gratings to generate the output signal beams.

Multichannel room: an enclosed space in which is configured to allow an absorptive interaction involving numerous analytes and light beams. Multichannel flow cells and capillary arrays can be located in a multichannel chamber.

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