Methods for quantitative and qualitative analyses of phospholipids using one-dimensional thin layer chromatography
First Claim
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1. A method for the simultaneous quantitative and qualitative determination of individual phospholipids in a phospholipid mixture, wherein said phospholipid mixture comprises a neutral lipid, said method comprising:
- dissolving said phospholipid mixture in at least one extraction solvent;
applying said at least one extraction solvent having said phospholipid mixture dissolved therein to a thin layer chromatography (TLC) plate;
placing said TLC plate having said phospholipid mixture applied thereto into an elution solvent mixture comprising chloroform, methanol, acetic acid, and an aqueous solution of potassium chloride; and
allowing said phospholipid mixture to migrate up said TLC plate in one direction until said individual phospholipids are resolved into discrete, detectable spots such that all said individual phospholipids present in said phospholipid mixture can be individually detected and said discrete detectable spots are separately quantifiable.
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Abstract
A highly sensitive and specific method for the detection and quantification of lipids is provided. Specifically, methods for the simultaneous detection and quantification of phospholipids extracted from mammalian tissues is described. The analytical methods provided disclose a modified one-dimensional thin-layer chromatography technique specifically developed to rapidly and accurantely detect and quantify phospholipids from mammalian cardiac tissues.
18 Citations
22 Claims
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1. A method for the simultaneous quantitative and qualitative determination of individual phospholipids in a phospholipid mixture, wherein said phospholipid mixture comprises a neutral lipid, said method comprising:
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dissolving said phospholipid mixture in at least one extraction solvent;
applying said at least one extraction solvent having said phospholipid mixture dissolved therein to a thin layer chromatography (TLC) plate;
placing said TLC plate having said phospholipid mixture applied thereto into an elution solvent mixture comprising chloroform, methanol, acetic acid, and an aqueous solution of potassium chloride; and
allowing said phospholipid mixture to migrate up said TLC plate in one direction until said individual phospholipids are resolved into discrete, detectable spots such that all said individual phospholipids present in said phospholipid mixture can be individually detected and said discrete detectable spots are separately quantifiable. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method for the quantitative determination of individual phospholipids in a phospholipid mixture, wherein said phospholipid mixture comprises a neutral lipid, said method comprising:
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dissolving said phospholipid mixture in at least one extraction solvent;
applying said at least one extraction solvent having said phospholipid mixture dissolved therein to a thin layer chromatography (TLC) plate;
placing said TLC plate having said phospholipid mixture applied thereto into an elution solvent mixture comprising chloroform, methanol, acetic acid, and an aqueous solution of potassium chloride; and
allowing said phospholipid mixture to migrate up said TLC plate in one direction until said individual phospholipids are resolved into discrete, detectable spots such that all said individual phospholipids present in said phospholipid mixture can be individually detected and said discrete detectable spots are stately quantifiable. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 20)
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21. A method for the simultaneous quantitative and qualitative determination of individual phospholipids in a phospholipid mixture, said method comprising:
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dissolving said phospholipid mixture in at least one extraction solvent;
applying said at least one extraction solvent having said phospholipid mixture dissolved therein to a thin layer chromatography (TLC) plate;
placing said TLC plate having said phospholipid mixture applied thereto into an elution solvent mixture comprising chloroform, methanol, acetic acid, and an aqueous solution of potassium chloride; and
allowing said phospholipid mixture to migrate up said TLC plate in one direction until said individual phospholipids are resolved into discrete, detectable spots such that all said individual phospholipids present in said phospholipid mixture can be individually detected and said discrete detectable spots are separately quantifiable. - View Dependent Claims (22)
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Specification