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US20040197235: Methods and apparatus for determining analytes in various matrices
67 Claims, 17 Drawings
Methods and apparatus for qualitatively or quantitatively determining one or more analytes in matrices such as foods, biological fluids, etc. An embodiment for determination of a single analyte comprises a sample receiving vessel, a first membrane and a reagent-containing well. The prepared sample passes through the first membrane whereby extraneous matter is removed, and a filtrate enters the reagent-containing well to provide a filtrate-reagent admixture from which the analyte may be determined. An embodiment for determination for multiple analytes includes one or more additional membranes in series with the first membrane, each such additional membrane being operative to capture one or more analytes. Each of the additional analytes may then be eluted from the membrane upon which it has been captured, into a separate reagent-containing well to provide eluant-reagent admixture from which each desired analyte may be determined. Formulations for preparation additives are also included. Additionally, theres provided an embodiment of above-described invention for determination of an analyte which is present in a matrix at low (e.g., sub-detectable) levels, wherein the filter of the apparatus is utilized to capture and concentrate the analyte, to provide a filtrate-reagent admixture wherein the analyte is present at detectable concentration.
- 1. An apparatus for non-electrophoretic determination of the presence of at least one analyte in each of n flowable samples, said apparatus comprising:
a housing having a cavity formed therein;
n filtrate-receiving vessels positioned within the cavity of said housing;
n membrane components, each of said membrane components being positioned in association with one of said filtrate-receiving vessels;
n sample-receiving wells, each of said sample-receiving wells being positioned in association with one of said membrane components such that sample placed within a particular sample receiving well may be caused to filter through the associated membrane component, and a filtrate which emerges from that membrane component will be received within the associated filtrate-receiving vessel;
a lid for sealing each of said sample receiving vessels and said cavity of said housing;
a differential pressure source to cause a pressure differential between each of said sample-receiving wells and each of said filtrate-receiving vessels, said pressure differential being operative to drive each sample through the associated membrane component and the resultant filtrate into the associated filtrate-receiving vessel.
- 18. A system for non-electrophoretic determination of at least a first analyte contained within a matrix, said system comprising:
a first membrane module having a membrane which is operative to prevent some of the matter of said matrix from passing therethrough, while allowing a filtrate containing said first analyte to pass therethrough;
a first vessel positioned in relation to said first membrane so as to receive said filtrate therein; and,
at least one reagent which is combinable with said filtrate in said receiving vessel to provide a reagent-filtrate admixture containing said first analyte and from which said first analyte may be determined.
- 44. A method for determining histamine in a sample, said method comprising the steps of:
A. adding to the sample a reagent which causes histamine to oxidize with resultant production of H2l O2; and, thereafter,
B. determining H2O2 in the sample as an indicator of histamine which was present prior to oxidation.
- 51. A method for determining free fatty acids in a sample, said method comprising the steps of:
A. adding a quantity of xylenol orange to the sample; and,
B. determining the change in color of the xylenol orange to indicate free fatty acids.
- 55. A method for determining free fatty acids in a sample, said method comprising the steps of:
A. adding a quantity of thymol blue to the sample; and,
B. determining the change in color of the thymol blue to indicate free fatty acids.
- 56. A method for determining lipid peroxides in a sample, said method comprising the steps of:
A. adding to the sample a quantity of hemoglobin and an activated electron donor substance, such that lipid peroxides present in the sample will cause at least some of the hemoglobin to convert to a modified hemoglobin derivative; and,
B. determining the amount of modified hemoglobin derivative present as an indication of lipid peroxides in the sample.
- 63. A method of determining sulfite and/or bisulfite in a sample, said method comprising the steps of:
A. adding a trivalent iron-xylenol orange complex to the sample; and,
B. determining the change in color of the trivalent iron-xylenol orange complex as an indicator of sulfite and/or bisulfite in the sample.
- 66. The method of claim be wherein step A is carried out by adding to the sample a reagent containing 0.1-5.0% by weight of Fe3(xanthine oxidase) in water/isopropanol solution.
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