Tacrolimus Extraction Using Aqueous Detergents

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Solution Overview

Problem

Current tacrolimus quantification methods rely on organic solvents, which are hazardous, volatile, and require manual, labor-intensive sample preparation, limiting automation and accuracy due to evaporation issues and human error.

Innovation Solution

A fully automated method using non-precipitating, non-denaturing aqueous solutions with bile salts or zwitterionic detergents like CHAPS to extract tacrolimus from blood samples, eliminating the need for organic solvents and centrifugation, and allowing for complete automation of the quantification process.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If organic solvents are used to extract tacrolimus from blood samples, then extraction efficiency is improved, but safety hazards and volatility increase

Engineering Contradiction:
Improveextraction efficiencyVSAvoidsafety hazards
Core Design Contradiction:
Measurement precisionVSObject-affected harmful factors

Solution Approach 1:

The patent changes the chemical parameter of the extraction medium from organic solvents (methylene chloride, ethyl acetate, methanol) to aqueous solutions containing non-ionic detergents (Triton X-100, Tween 80, Brij 35). This parameter change maintains extraction efficiency while eliminating the harmful properties of organic solvents including flammability, toxicity, and excessive volatility.

Inventive Principle:
Principle #35Parameter changes

2Measurement precision

If organic solvents are used for tacrolimus extraction, then extraction effectiveness is improved, but automation is limited due to evaporation interference

Engineering Contradiction:
Improveextraction effectivenessVSAvoidautomation capability
Core Design Contradiction:
Measurement precisionVSExtent of automation

Solution Approach 1:

The patent changes the physical parameter of the extraction medium from volatile organic solvents to non-volatile aqueous solutions. This eliminates evaporation interference during automated processing steps, allowing complete automation of the tacrolimus quantification assay without manual intervention for solvent handling or evaporation control.

Inventive Principle:
Principle #35Parameter changes

3Ease of manufacture

If manual sample preparation procedures are used, then extraction can be performed, but labor intensity and time consumption increase

Engineering Contradiction:
Improveextraction capabilityVSAvoidanalytical throughput
Core Design Contradiction:
Ease of manufactureVSProductivity

Solution Approach 1:

The patent merges multiple separate manual steps (denaturation, extraction, centrifugation, decantation) into a single automated liquid handling process using aqueous detergent solutions. This consolidation enables complete automation, dramatically increasing analytical throughput and eliminating manual labor while maintaining extraction capability.

Inventive Principle:
Principle #5Merging (Combining)

4Measurement precision

If centrifugation and decantation steps are included, then separation of extracted sample is achieved, but process complexity increases

Engineering Contradiction:
Improvesample separationVSAvoidprocedure complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent extracts and removes the need for centrifugation and decantation steps by using aqueous detergent-based extraction that is compatible with automated liquid handling systems. The extraction process itself enables direct transfer of the supernatant without requiring separate separation steps, thereby reducing procedural complexity while maintaining sample separation quality.

Inventive Principle:
Principle #2Taking out (Extraction)

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 method provides accurate and reproducible tacrolimus quantification with reduced costs and improved safety by eliminating manual handling and volatile solvents, achieving results comparable to traditional methods while ensuring automation and minimizing interference with the quantification process.

Implementation Method 1

In order to determine the presence of Tacrolimus and its concentration in a human blood sample, RBC's present in the sample must be lysed to release the tacrolimus/protein complexes

Methodology Applied
Scientific EffectHemolysis:

Implementation Method 2

A fully automated method using non-precipitating, non-denaturing aqueous solutions with bile salts or zwitterionic detergents like CHAPS to extract tacrolimus from blood samples

Methodology Applied
Scientific EffectSolubilization: Solvation

Data Source

PatentEP1949100B1Method of tacrolimus extraction and quantification using aqueous detergents
Publication Date: 2011.08.03 ABBOTT LAB INC
  • EP1949100B1 patent drawingFigure 1
  • EP1949100B1 patent drawingFigure 2
  • EP1949100B1 patent drawingFigure 3

AI summary

The present invention relates to a method of extracting tacrolimus in a test blood sample to use in a tacrolimus quantification assay. The method of the present invention extracts tacrolimus in non-precipitating, non-denaturing aqueous environment, avoiding cumbersome manual pretreatment procedures or the use of organic solvents. The method of the present invention permits full automation of tacrolimus quantification assays.