Method of assaying whole blood for HDL cholesterol

Claim 1

A method of separating cellular components, low density lipoprotein fraction and very low density lipoprotein fraction from an undiluted whole blood sample consisting essentially of: (a) first directly contacting the undiluted whole blood sample with a first carrier matrix to separate the cellular components of the whole blood sample and provide a plasma or serum, wherein said first carrier matrix has incorporated therein about 25 to about 250 units of an agglutinin, about 50 to about 150 NIH units of a coagulant or a mixture thereof to agglutinize or coagulate the cellular components of the undiluted whole blood sample in the first carrier matrix; (b) then allowing the plasma or serum to migrate by gravity to a second carrier matrix that has the same dimensions and is in fluid communication with the first carrier matrix to separate the low density lipoprotein fraction and the very low density lipoprotein fraction from the plasma or serum, said second carrier matrix having a pore size of at least 1 to about 50 microns and having incorporated therein a precipitating reagent composition comprising a sufficient amount of a polyvalent metal ion and a sufficient amount of a precipitating compound to separate the low density lipoprotein fraction and the very low density lipoprotein fraction from the plasma or serum and to provide a serum or plasma including a high density lipoprotein fraction, wherein the precipitating compound is selected from the group consisting of dextran sulfate, heparin sodium, phosphotungstic acid, polyvinyl sulfate, sodium tungstate, ammonium molybdate, and mixtures thereof; and (c) then allowing the serum or plasma including the high density lipoprotein fraction to flow by gravity through the second carrier matrix.

Claim 2

A method of separating cellular components, low density lipoprotein fraction and very low density lipoprotein fraction from an undiluted whole blood sample consisting essentially of: (a) first directly contacting the undiluted whole blood sample with a first carrier matrix to separate the cellular components of the whole blood sample and provide a plasma or serum, wherein said first carrier matrix has incorporated therein about 25 to about 250 units of an agglutinin, about 50 to about 150 NIH units of a coagulant or a mixture thereof to agglutinize or coagulate the cellular components of the undiluted whole blood sample in the first carrier matrix; (b) then allowing the plasma or serum to migrate by gravity to a second carrier matrix that has the same dimensions and is in fluid communication with the first carrier matrix to separate the low density lipoprotein fraction and the very low density lipoprotein fraction from the plasma or serum, said second carrier matrix having a pore size of at least 1 to about 50 microns and having incorporated therein a precipitating reagent composition comprising a sufficient amount of a polyvalent metal ion and a sufficient amount of a precipitating compound to separate the low density lipoprotein fraction and the very low density lipoprotein fraction from the plasma or serum and to provide a serum or plasma including a high density lipoprotein fraction, wherein the precipitating compound is selected from the group consisting of dextran sulfate, heparin sodium, phosphotungstic acid, polyvinyl sulfate, sodium tungstate, ammonium molybdate, and mixtures thereof; and (c) then allowing the serum or plasma including the high density lipoprotein fraction to flow by gravity through the second carrier matrix.

Claim 6

A method of separating plasma or serum that is essentially free of cellular components, low density lipoproteins and very low density lipoproteins, from an undiluted whole blood sample and assaying the plasma or serum for a predetermined constituent of high density lipoproteins present in the plasma or serum consisting essentially of: (a) first directly contacting the undiluted whole blood sample with a first carrier matrix to separate the cellular components of the whole blood sample and to provide a plasma or serum, wherein said first carrier matrix has incorporated therein a separating reagent composition comprising about 25 to about 250 units an agglutinin, about 50 to about 150 NIH units of a coagulant or a mixture thereof to agglutinize or coagulate the cellular components of the undiluted whole blood sample in the first carrier matrix; (b) then allowing the plasma or serum to migrate by gravity to a second carrier matrix that has the same dimensions and is in fluid communication with the first carrier matrix to separate the low density lipoproteins and the very low density lipoproteins from the plasma or serum, said second carrier matrix having a pore size of at least 1 to about 50 microns and having incorporated therein a precipitating reagent comprising a sufficient amount of a polyvalent metal ion and a sufficient amount of a precipitating compound to separate the low density lipoproteins and the very low density lipoproteins from the plasma or serum and to provide a serum or plasma that is essentially free of cellular components, low density lipoproteins and very low density lipoproteins and that includes high density lipoproteins, wherein the precipitating compound is selected from the group consisting of dextran sulfate, heparin sodium, phosphotungstic acid, polyvinyl sulfate, sodium tungstate, ammonium molybdate, and mixtures thereof; (c) then allowing the serum or plasma including the high density lipoproteins to flow by gravity through the second carrier matrix to contact a test area incorporating a reagent composition to assay for the predetermined constituent of the high density lipoproteins in the plasma or serum, said reagent composition capable of interacting with the predetermined constituent of the high density lipoprotein to produce a detectable change in the test area; and (d) examining the test area for a detectable change, wherein the assay is independent of the amount of hematocrit present in the whole blood sample.

Claim 7

A method of separating plasma or serum that is essentially free of cellular components, low density lipoproteins and very low density lipoproteins, from an undiluted whole blood sample and assaying the plasma or serum for a predetermined constituent of high density lipoproteins present in the plasma or serum consisting essentially of: (a) first directly contacting the undiluted whole blood sample with a first carrier matrix to separate the cellular components of the whole blood sample and to provide a plasma or serum, wherein said first carrier matrix has incorporated therein a separating reagent composition comprising about 25 to about 250 units an agglutinin, about 50 to about 150 NIH units of a coagulant or a mixture thereof to agglutinize or coagulate the cellular components of the undiluted whole blood sample in the first carrier matrix; (b) then allowing the plasma or serum to migrate by gravity to a second carrier matrix that has the same dimensions and is in fluid communication with the first carrier matrix to separate the low density lipoproteins and the very low density lipoproteins from the plasma or serum, said second carrier matrix having a pore size of at least 1 to about 50 microns and having incorporated therein a precipitating reagent comprising a sufficient amount of a polyvalent metal ion and a sufficient amount of a precipitating compound to separate the low density lipoproteins and the very low density lipoproteins from the plasma or serum and to provide a serum or plasma that is essentially free of cellular components, low density lipoproteins and very low density lipoproteins and that includes high density lipoproteins, wherein the precipitating compound is selected from the group consisting of dextran sulfate, heparin sodium, phosphotungstic acid, polyvinyl sulfate, sodium tungstate, ammonium molybdate, and mixtures thereof; (c) then allowing the serum or plasma including the high density lipoproteins to flow by gravity through the second carrier matrix to contact a test area incorporating a reagent composition to assay for the predetermined constituent of the high density lipoproteins in the plasma or serum, said reagent composition capable of interacting with the predetermined constituent of the high density lipoprotein to produce a detectable change in the test area; and (d) examining the test area for a detectable change, wherein the assay is independent of the amount of hematocrit present in the whole blood sample.