DEVELOPMENT OF THE METHOD FOR DETERMINING THE CONTENT OF THE SYNTHETIC DYE DIAMOND BLUE FCF IN THE SORPTION MATERIAL BY THE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD
At a Glance
Section titled “At a Glance”| Metadata | Details |
|---|---|
| Publication Date | 2023-04-27 |
| Journal | Medical and Clinical Chemistry |
| Authors | А.А. Borysenko, А. М. Антоненко, S. M. Holoborodko, K. P. Antonyuk, D. S. Milokhov |
| Citations | 1 |
| Analysis | Full AI Review Included |
Executive Summary
Section titled “Executive Summary”This research details the development and validation of a High-Performance Liquid Chromatography (HPLC) method for quantifying the synthetic tracer dye, Diamond Blue FCF (DB FCF), within sorption materials (filter paper).
- Core Value Proposition: The method provides a necessary analytical tool for evaluating the effectiveness and safety of Unmanned Aerial Vehicles (UAVs) used in agricultural pesticide application, supporting hygienic regulation and risk assessment.
- Analytical Technique: The methodology relies on reversed-phase HPLC coupled with spectrophotometric (SP) detection, optimized for high sensitivity and selectivity.
- Sample Preparation: A multi-stage preparation process includes aqueous extraction, pH adjustment (to pH < 2), and subsequent Solid Phase Extraction (SPE) cleanup using a Strata™ C18-e cartridge.
- Performance Metrics: The method reliably measures DB FCF content in the range of 0.125 to 1.25 µg/dm2 of sorption material.
- Chromatographic Precision: Under optimized conditions (30 °C column temperature, 1.0 mL/min flow rate), DB FCF exhibits a consistent retention time of 5.5 ± 0.1 minutes.
- Elution Chemistry: The dye is eluted from the C18 sorbent using a 3 mL solution of 2% (v/v) aqueous ammonia in methanol, followed by concentration via rotary evaporation (max 40 °C).
Technical Specifications
Section titled “Technical Specifications”| Parameter | Value | Unit | Context |
|---|---|---|---|
| Measurement Range | 0.125 to 1.25 | µg/dm2 | DB FCF content in sorption material |
| HPLC Column Type | Microsorb 100-5 C18 | N/A | Steel, 150/4.6 mm dimensions |
| Pre-column Type | Microsorb 100-5 C18 | N/A | Steel, 4/3 mm dimensions |
| Column Temperature | 30 | °C | HPLC thermostat setting |
| Detection Wavelength | 620 | nm | Diode Array Detector (DAD) setting |
| Mobile Phase Flow Rate | 1.0 | mL/min | Volumetric flow rate |
| Injection Volume | 10 | µL | Sample volume for HPLC |
| Retention Time (tR) | 5.5 ± 0.1 | minutes | Diamond Blue FCF |
| SPE Cartridge Specification | C18-e (55 µm) | 500 mg/6 mL | Strata™ cartridge used for cleanup |
| Rotary Evaporator Temp | Max 40 | °C | Water bath temperature during concentration |
| DB FCF Molecular Formula | C37H34Na2N2O9S3 | N/A | Empiric formula |
| DB FCF Molecular Weight | 792.9 | g/mol | Calculated weight |
Key Methodologies
Section titled “Key Methodologies”The analytical determination of Diamond Blue FCF (DB FCF) in the sorption material (filter paper) follows a rigorous, multi-step protocol:
- Aqueous Extraction: The sorption material (2 dm2 filter paper) is crushed and subjected to initial extraction using 50 mL of distilled water, followed by vigorous hand shaking for 1 minute. A subsequent wash with 10 mL of distilled water is added to the main extract.
- pH Adjustment: The combined aqueous extract is acidified by adding 1 M aqueous sulfuric acid (H2SO4) until the pH value is set to less than or equal to 2, verified using indicator paper.
- Solid Phase Extraction (SPE): The acidified extract is applied to a prepared Strata™ C18-e cartridge (55 µm, 70 °C). The cartridge is then washed with 10 mL of bidistilled water, and the wash solvents are discarded.
- Elution and Concentration: DB FCF is eluted from the sorbent using 3 mL of 2% (v/v) aqueous ammonia solution in methanol. The eluate is collected and concentrated via rotary evaporation (water bath temperature not exceeding 40 °C) to a volume of 0.2-0.3 mL, followed by complete evaporation in air to a dry residue.
- Final Sample Preparation: The dry residue is dissolved in 1 mL of bidistilled water, yielding the final sample extract volume of 1 mL for chromatographic injection.
- HPLC Analysis: Chromatography is performed using a gradient mobile phase system: Eluent A (Acetonitrile) and Eluent B (0.1% aqueous orthophosphoric acid). The analysis uses a steel C18 column maintained at 30 °C, with a flow rate of 1.0 mL/min, and detection is performed at 620 nm.
Commercial Applications
Section titled “Commercial Applications”The developed HPLC methodology is critical for industries and regulatory bodies involved in advanced agricultural practices and environmental safety monitoring.
- Precision Agriculture Technology: Essential for validating the efficiency and coverage of pesticide delivery systems utilizing Unmanned Aerial Vehicles (UAVs).
- Agrochemical Regulation: Supports the detailed study, risk assessment, and hygienic regulation of new pesticide application technologies, particularly during pre-registration trials and scientific research.
- Environmental Safety and Toxicology: Used to quantify the loss of pesticide formulation outside the treated field, minimizing negative environmental and population health impacts.
- Tracer Studies: Applicable in environmental engineering and hydrology where DB FCF is used as a standard dye tracer for monitoring solute transport and flow paths in soil or water systems.
- Quality Control (QC) in Farming: Provides a standardized method for evaluating the performance of various UAV models and pesticide combinations under different agrotechnical conditions.
View Original Abstract
Introduction. To date, the use of unmanned aerial vehicles (UAVs) in Ukraine for the agricultural lands treatment is a new promising technology that requires detailed study and development of approaches to risk assessment and hygienic regulation. Development of a method that will allow measuring the content of diamond blue FCF in the sorption material (filter paper) is relevant. The aim of the study - development of the method for determining the content of the synthetic dye diamond blue FCF in the sorption material by the high-performance liquid chromatography method. Research Methods. The following materials were used: laboratory analytical balance Radwag® AS220.R2, rotary evaporator, cartridge Strata™ C18-e (55 μm, 70 °С) 500 mg/6 ml, f. Phenomenex, steel chromatographic column 150/4.6 Microsorb 100-5 C18, pre-column chromatographic 4/3 Microsorb 100-5 C18, liquid chromatograph “Shimadzu” with a diode array detector, Diamond blue FCF, standard, 87.7, paper filters de-ashed “red ribbon”. Results and Discussion. At the first stage of the research, samples were taken and prepared. For analysis, 2 parallel samples were taken. The next stage was the preparation of the sample for introduction into the chromatograph. The third stage was chromatography performing. At the final stage, the diamond blue FCF peak areas were determined and calculated on the chromatograms. The indicated method of determining the content of diamond blue FCF in the sorption material (filter paper) includes extraction of the dye from the sorption material (filter paper) with distilled water; solid-phase extraction and quantification of diamond blue FCF by reversed-phase HPLC with SF detection. This method differs from the known ones in that it makes it possible to determine the investigated dye in the sorption material. Conclusion. The proposed method of containing diamond blue FCF in the sorption material (filter paper) will allow to evaluate the effectiveness and safety of the use of various models of UAVs in combination with various pesticide preparations when using different agrotechnical characteristics at the stage of pre-registration trials and scientific research.