Determination of Tyrian purple by high performance liquid chromatography with diode array detection
Athina Vasileiadou1, Ioannis Karapanagiotis2, Anastasia Zotou1
1Laboratory of Analytical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
Journal of Chromatography. A
|April 30, 2016
Summary
This study quantifies Tyrian purple components like indigotin and indirubin using an efficient HPLC method. The validated method accurately measures these compounds in mollusc samples, revealing concentrations up to 12.2 μg/mg.
Area of Science:
- Analytical Chemistry
- Marine Biology
- Natural Products Chemistry
Background:
- Tyrian purple, a historically significant dye, is derived from the hypobranchial glands of marine molluscs.
- The precise quantification of its constituent compounds, including indigotin and its brominated derivatives, is crucial for chemical and historical analysis.
- Previous analytical methods may lack the efficiency or sensitivity required for complex natural matrices.
Purpose of the Study:
- To develop and validate an efficient High-Performance Liquid Chromatography (HPLC) method for quantifying indigotin, indirubin, and their brominated derivatives.
- To analyze Tyrian purple components in samples from Hexaplex (Murex) trunculus molluscs collected from Tunisian and Croatian coastlines.
- To assess the accuracy and reliability of the developed HPLC method for natural product analysis.
Main Methods:
- Quantification of indigotin, indirubin, 6-bromoindigotin, 6'-bromoindirubin, 6-bromoindirubin, 6,6'-dibromoindigotin, and 6,6'-dibromoindirubin using HPLC with a diode array detector.
- Separation achieved via gradient elution on a C18 reversed-phase column at 35°C with a mobile phase of water and acetonitrile containing trifluoroacetic acid.
- Method validation included linearity, detection/quantification limits, precision, accuracy, ruggedness, and robustness; analysis of mollusc samples employed the standard addition approach.
Main Results:
- The validated HPLC method demonstrated limits of detection between 0.02-0.05 μg/mL and linearity up to 2.5 μg/mL for all analytes.
- Analysis of four Hexaplex (Murex) trunculus samples revealed concentrations of indigotin, indirubin, and their brominated derivatives ranging from 0.01 to 12.2 μg/mg.
- The standard addition approach and recovery experiments confirmed the method's accuracy and suitability for analyzing these compounds in mollusc extracts.
Conclusions:
- An efficient and validated HPLC-diode array detector method enables accurate quantification of Tyrian purple components in mollusc samples.
- The study successfully identified and quantified key indigotin and indirubin derivatives in Hexaplex (Murex) trunculus from the Mediterranean.
- This analytical scheme provides a reliable tool for the chemical characterization of Tyrian purple and related natural colorants.
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