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US 11,674,965 B2Granted
Method for preparing and analyzing fluorescent compounds in plasma
AssigneeMediBeacon Inc.
InventorJeng J. Shieh
Life of this patent
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Abstract
Disclosed herein is a method for analyzing the concentration of a fluorescent compound in the plasma of a patient. The method includes collecting a sample of plasma from a patient, diluting the sample with a solvent and analyzing the diluted sample by HPLC. The sample does not need to be dried down during sample preparation nor is an internal standard required.
Claims23
A method for measuring the amount of a fluorescent compound in plasma, the method comprising:
- diluting a sample of plasma comprising the fluorescent compound with an aqueous buffer;
- and
- analyzing the diluted sample by high performance liquid chromatography (HPLC) thereby measuring the amount of the fluorescent compound in the sample of plasma;
- wherein the sample of plasma is not dried before HPLC analysis, and no internal standard is added to the sample,
- wherein the method does not include a protein precipitation step that comprises causing proteins to precipitate from the sample of plasma and removing the precipitated proteins from the sample prior to HPLC analysis,
- wherein the fluorescent compound is a compound of Formula I, or a pharmaceutically acceptable salt thereof, wherein
- each of X1 and X2 is independently —CO2R1, —CONR1R2, —CO(AA) or —CONH(PS);
- each of Y1 and Y2 is independently selected from the group consisting of —NR1R2;
- and
- Z1 is a single bond, —CR1R2—, —O—, —NR1—, —NCOR1—, —S—, —SO—, or —SO2—;
- each of R1 to R2 are independently selected from the group consisting of H, —CH2(CHOH)aH, —CH2(CHOH)aCH3, —CH2(CHOH)aCO2H, —(CHCO2H)aCO2H, —(CH2CH2O)cH, —(CH2CH2O)cCH3, —(CH2)aSO3H, —(CH2)aSO3−, —(CH2)aSO2H, —(CH2)aSO2−, —(CH2)aNHSO3H, —(CH2)aNHSO3−, —(CH2)aNHSO2H, —(CH2)aNHSO2−, —(CH2)aPO3H2, —(CH2)aPO3H−, and —(CH2)aPO32−;
- AA is a peptide chain comprising one or more amino acids selected from the group consisting of natural and unnatural amino acids, linked together by peptide or amide bonds and each instance of AA may be the same or different than each other instance;
- PS is a sulfated or non-sulfated polysaccharide chain comprising one or more monosaccharide units connected by glycosidic linkages;
- and
- ‘a’ is a number from 1 to 10, ‘c’ is a number from 1 to 100, and each of ‘m’ and ‘n’ are independently a number from 1 to 3.
12 more elements
A method for measuring the amount of a fluorescent compound in plasma, the method comprising:
- diluting a sample of plasma comprising the fluorescent compound with an aqueous buffer comprising a phosphate buffering agent, wherein the sample of plasma is diluted with the aqueous buffer in a dilution ratio (v/v) of from 1:5 to 1:1000 (sample:buffer);
- and
- analyzing the diluted sample by high performance liquid chromatography (HPLC) thereby measuring the amount of the fluorescent compound in the sample of plasma;
- wherein the sample of plasma is not dried before HPLC analysis, and no internal standard is added to the sample,
- wherein the method does not include a protein precipitation step that comprises causing proteins to precipitate from the sample of plasma and removing the precipitated proteins from the sample prior to HPLC analysis,
- wherein the fluorescent compound is a compound of Formula I, or a pharmaceutically acceptable salt thereof, wherein
- each of X1 and X2 is independently —CO2R1, —CONR1R2, —CO(AA) or —CONH(PS);
- each of Y1 and Y2 is independently selected from the group consisting of —NR1R2;
- and
- Z1 is a single bond, —CR1R2—, —O—, —NR1—, —NCOR1—, —S—, —SO—, or —SO2—;
- each of R1 to R2 are independently selected from the group consisting of H, —CH2(CHOH)aH, —CH2(CHOH)aCH3, —CH2(CHOH)aCO2H, —(CHCO2H)aCO2H, —(CH2CH2O)cH, —(CH2CH2O)cCH3, —(CH2)aSO3H, —(CH2)aSO3−, —(CH2)aSO2H, —(CH2)aSO2−, —(CH2)aNHSO3H, —(CH2)aNHSO3−, —(CH2)aNHSO2H, —(CH2)aNHSO2−, —(CH2)aPO3H2, —(CH2)aPO3H−, and —(CH2)aPO32−;
- AA is a peptide chain comprising one or more amino acids selected from the group consisting of natural and unnatural amino acids, linked together by peptide or amide bonds and each instance of AA may be the same or different than each other instance;
- PS is a sulfated or non-sulfated polysaccharide chain comprising one or more monosaccharide units connected by glycosidic linkages;
- and
- ‘a’ is a number from 1 to 10, ‘c’ is a number from 1 to 100, and each of ‘m’ and ‘n’ are independently a number from 1 to 3.
12 more elements
Dependencies are inferred from the claim text.