Fluorescent Compound Plasma Analysis via Direct HPLC
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Solution Overview
Problem
Current methods for assessing renal function, such as serum creatinine levels and exogenous GFR tracer agents, are labor-intensive, time-consuming, and lack sensitivity and accuracy, particularly for real-time kidney function monitoring in patients with chronic kidney disease.
Innovation Solution
A method involving the collection and analysis of plasma samples using high-pressure liquid chromatography (HPLC) without drying the samples or adding an internal standard, allowing for direct dilution or protein precipitation followed by HPLC analysis to measure the concentration of fluorescent compounds like MB-102, which emits a strong fluorescent signal for real-time GFR determination.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional protein precipitation and drying methods are used before HPLC analysis, then sample preparation is more thorough, but analysis time increases and productivity decreases
Solution Approach 1:
The invention extracts only the essential step for accurate measurement (protein precipitation) while eliminating unnecessary steps (drying and reconstitution). The plasma sample is directly analyzed by HPLC after protein precipitation, removing the time-consuming drying and reconstitution steps while maintaining measurement accuracy through direct injection of the precipitated sample.
Solution Approach 2:
The invention skips the traditional drying and reconstitution steps in the sample preparation process. By directly injecting the protein-precipitated plasma sample into HPLC, the method rushes through the analysis process efficiently, reducing analysis time from hours to minutes while maintaining sufficient precision for clinical measurements.
2Measurement precision
If multiple time-point blood samples are collected for GFR determination using iohexol, then GFR assessment accuracy is improved, but time consumption and operational complexity increase
Solution Approach 1:
The invention uses MB-102, a fluorescent compound with rapid renal clearance, allowing GFR determination from a single blood sample rather than multiple time-point collections. The preliminary selection of an optimized tracer agent enables accurate GFR measurement without the need for repeated sampling and complex pharmacokinetic modeling.
Solution Approach 2:
The invention replaces the complex mechanical process of multiple blood collections and laboratory-based PK analysis with a simplified fluorescent detection system. MB-102's strong fluorescence signal allows direct measurement of plasma concentration, eliminating the need for complex WinNonlin PK/PD modeling and reducing operational complexity.
3Measurement precision
If internal standards are added to plasma samples, then measurement accuracy is improved, but sample preparation complexity and error potential increase
Solution Approach 1:
The invention uses the endogenous plasma components and the fluorescent properties of MB-102 itself for self-quantification. The method relies on the compound's inherent fluorescence characteristics and comparison with external calibration curves, eliminating the need for additional internal standard substances and simplifying the sample preparation procedure.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach significantly reduces analysis time, minimizes errors, and provides accurate, real-time assessment of kidney function, enabling faster and more reliable monitoring of renal impairment.
Implementation Method 1
MB-102, a fluorescent compound that exhibits excellent photo-physical properties... MB-102... emits a strong fluorescent signal at 556 nm when excited 434 nm
Implementation Method 2
analyzing the diluted sample by HPLC thereby measuring the amount of the compound in the plasma
Data Source
AI summary
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.