DEVELOPMENT AND VALIDATION OF A GC/MS-BASED ASSAY FOR DETECTING PESTICIDE RESIDUES IN HUMAN BLOOD

Аннотация

INTRODUCTION samples is crucial for assessing exposure levels, The presence of pesticide residues in human blood understanding health impacts, and ensuring is a significant concern due to the potential health regulatory compliance. Gas Chromatography-Mass risks associated with long-term exposure. Spectrometry (GC/MS) has emerged as a powerful Pesticides, while essential for agricultural analytical tool for this purpose, offering high productivity, can have detrimental effects on sensitivity and specificity in the analysis of complex human health, including neurological disorders, mixtures. cancer, and endocrine disruption. Accurate and reliable detection of pesticide residues in biological Despite advancements in analytical techniques, https://www.theamericanjournals.com/index.php/tajmspr 7 THE USA JOURNALS THE AMERICAN JOURNAL OF MEDICAL SCIENCES AND PHARMACEUTICAL RESEARCH (ISSN – 2689-1026) VOLUME 06 ISSUE10 existing methods for detecting pesticide residues METHOD often face limitations, such as insufficient This study outlines the development and validation sensitivity, high detection limits, or interference of a novel Gas Chromatography-Mass from other blood components. To address these Spectrometry (GC/MS)-based assay for detecting challenges, this study focuses on the development pesticide residues in human blood. The and validation of a novel GC/MS-based assay methodology encompasses several key stages, designed to enhance the accuracy and reliability of including sample collection and preparation, assay pesticide residue detection in human blood. The development, and validation processes. assay aims to overcome the limitations of current methods by optimizing sample preparation and Blood samples were collected from consenting chromatographic conditions to improve detection volunteers under controlled conditions to ensure limits and minimize interference. consistency and minimize contamination. The collected samples were immediately processed to The development process involves refining the prevent degradation of pesticide residues. Initial extraction and clean-up procedures to effectively sample preparation involved centrifugation to isolate pesticide residues from blood matrices, separate plasma from cellular components. To followed by the optimization of GC/MS parameters isolate pesticide residues from the plasma, a liquid- to achieve precise separation and identification. liquid extraction (LLE) method was employed Validation of the assay is conducted to ensure its using a suitable organic solvent (e.g., hexane or robustness, including assessments of specificity, ethyl acetate). This extraction was optimized for sensitivity, linearity, and reproducibility. By efficiency and recovery rates. Following extraction, providing a more effective analytical tool for the samples underwent a clean-up process using monitoring pesticide exposure, this study solid-phase extraction (SPE) to remove potential contributes to the broader field of environmental interfering substances and concentrate the target and clinical toxicology, offering valuable insights analytes. into the risks associated with pesticide use and supporting efforts to safeguard public health. achieve high recovery and minimal interference. The development of the GC/MS-based assay Various solvents and SPE materials were tested to involved optimizing several parameters to enhance determine the most effective combination for analytical performance. The first step was to isolating pesticide residues from the blood matrix. optimize the extraction and clean-up procedures to https://www.theamericanjournals.com/index.php/tajmspr 8 THE USA JOURNALS THE AMERICAN JOURNAL OF MEDICAL SCIENCES AND PHARMACEUTICAL RESEARCH (ISSN – 2689-1026) VOLUME 06 ISSUE10 After sample preparation, the GC/MS conditions based on its ability to provide adequate separation were optimized, including the choice of column, of pesticide residues, while the MS parameters temperature program, and mass spectrometric were tuned to maximize sensitivity and specificity detection parameters. The GC column was selected for each target analyte. and calibration curves were constructed to verify The assay was rigorously validated according to the assay’s ability to produce accurate and standard guidelines to ensure its accuracy and consistent results. Reproducibility was assessed reliability. Key validation parameters included through repeated analyses of identical samples specificity, sensitivity, linearity, and under different conditions and by different reproducibility. Specificity was assessed by operators to confirm the assay’s reliability and evaluating the assay’s ability to distinguish precision. pesticide residues from other potential contaminants in the blood matrix. Sensitivity was The GC/MS data were analyzed using appropriate determined by establishing the limit of detection software to identify and quantify pesticide residues (LOD) and limit of quantification (LOQ) for each b