HPLC-ELSD Detection of Saccharides in Compound Kushen Injection
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Solution Overview
Problem
Current methods for detecting saccharides in Compound Kushen Injection, such as HPLC-ELSD and HPLC-CAD, face issues like poor separation between fructose and D-glucose anhydrous, excessive interference, and unstable baselines, making it difficult to accurately determine the content and fingerprint of sugar components.
Innovation Solution
A high-performance liquid chromatography-evaporative light scattering detection method using a Prevail Carbo hydrogen ES column with a gradient solution of acetonitrile and water, optimized elution conditions, and specific detector settings to simultaneously detect D-glucose anhydrous, D-fructose, sucrose, and pinitol, allowing for accurate content and fingerprint analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If HPLC-ELSD method is used to detect saccharide components, then the detection can be performed, but the separation degree between fructose and D-glucose anhydrous is relatively small and pinitol cannot be separated
Solution Approach 1:
The patent applies parameter changes by optimizing the mobile phase composition (acetonitrile-water gradient elution), column temperature (30°C), and flow rate (1.0 mL/min) to achieve complete separation of fructose, pinitol, and D-glucose anhydrous peaks, resolving the separation issue while maintaining detection capability
Solution Approach 2:
The patent uses dynamic gradient elution where the acetonitrile concentration increases over time (from 25% to 40% over 40 minutes), allowing dynamic optimization of separation conditions during the chromatographic run to resolve multiple saccharide components with different polarities
2Measurement precision
If HPLC-CAD method is used with Xbridge Amide chromatographic column, then monosaccharides and disaccharides can be detected, but there is excessive interference from other substances and unstable baseline
Solution Approach 1:
The patent introduces an intermediary substance (oxalic acid) in the mobile phase that suppresses ionization of saccharide components, reducing interference from other substances and stabilizing the baseline in the CAD detector
Solution Approach 2:
The patent changes the mobile phase parameters by adding oxalic acid and using acetonitrile-water gradient elution, which improves peak shape, reduces interference, and stabilizes the baseline while maintaining detection sensitivity
3Manufacturing precision
If Prevail Carbohydrate ES column with acetonitrile-water mobile phase is used, then fructose and pinitol can be separated, but the method complexity increases
Solution Approach 1:
The patent achieves multi-functionality by using a single Prevail Carbohydrate ES column and acetonitrile-water mobile phase system that can simultaneously separate multiple types of saccharides (monosaccharides and disaccharides) without requiring multiple different columns or complex sample preparation steps
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 a fast and efficient means to simultaneously determine the content and fingerprint of multiple sugar components in Compound Kushen Injection, overcoming previous challenges of separation and interference, and ensuring accurate quality control.
Implementation Method 1
high-performance liquid chromatography-evaporative light scattering detection method
Implementation Method 2
evaporative light scattering detection method
Data Source
AI summary
A method for detecting the monosaccharide and oligosaccharide content and fingerprint spectrum of a compound Sophora flavescens injection, comprising: carrying out detection by using a high-performance liquid chromatography-evaporation light scattering detection method, wherein the monosaccharides and oligosaccharides are D-anhydrous glucose, D-fructose, sucrose and pinitol. The present method is an improved method for detecting monosaccharides and oligosaccharides of a compound Sophora flavescens injection, and can measure four saccharide components in the compound Sophora flavescens injection at the same time, and constructs an HPLC-ELSD fingerprint spectrum to provide a technical method for the quality control of the saccharide components in the compound Sophora flavescens injection.