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Optimizing the Concentration of Nile Red for Screening of Microplastics in Bottled Water
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Scientists developed a better way to detect tiny plastic particles in bottled water by fine-tuning a special dye test, making it more accurate at spotting microplastics of different sizes. When they tested seven brands of bottled water (still and sparkling, in both plastic and glass bottles), they confirmed that a majority of the flagged particles were indeed plastic, a useful step toward more reliable methods for tracking how much plastic we might be drinking.
Increasing concern regarding the presence of microplastics in drinking water has led to a growing number of studies aimed at quantifying microplastics in water.In this work, we present an optimized procedure for the use of Nile red (NR) as a fluorescent staining agent for pre-screening of microplastics in bottled water.Positive and negative control experiments with NR concentrations ranging from 0.001 to 10 mg/L showed that the appropriate NR concentration is an important factor in obtaining representative particle counts.Non-optimized staining concentrations led to underestimation or overestimation of the particle count.In this study, the optimized NR staining concentration was found to be 0.1 mg/L.This method was successfully used to screen particles in seven different brands of bottled water consisting of both still and carbonated water, in both plastic and glass bottles.Particles larger than 100 µm were chemically characterized using attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR).Sixtyseven percent of these particles pre-screened with NR were confirmed to be polymers.Particles smaller than 100 µm were qualitatively analyzed using pyrolysis coupled with gas chromatography and mass spectroscopy (Py-GC-MS).Analysis of polymers between ~5-100 µm using Py-GC-MS confirmed that this smaller fraction generally mirrors the FTIR results for particles larger than l00 µm.
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Optimizing the Concentration of Nile Red for Screening of Microplastics in Drinking Water
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This study optimized the concentration of Nile red fluorescent dye for pre-screening microplastics in drinking water, establishing a protocol that balances detection sensitivity with background fluorescence to improve accuracy in identifying plastic particles.
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Researchers developed a novel method for counting and sizing microplastic particles in bottled water using Nile Red fluorescent staining combined with direct microscopic counting. Testing Iranian bottled water brands revealed widespread microplastic contamination, with the new method providing a faster and more accessible alternative to spectroscopic identification for routine bottled water monitoring.
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Researchers refined a Nile Red dye-based method to count nanoplastic particles in water by reducing dye concentration to near-nanomolar levels, minimizing false positives from dye aggregates that mimic plastic particles. Using this approach they detected approximately 250 nanoplastics per nanoliter in water from plastic bottles and cartons — roughly ten times more than in tap water.
Efficient and Scalable Detection of Microplastics in Drinking Water Using Fluorescence High‐Content Imaging
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Researchers developed a rapid, high-throughput fluorescence imaging method using Nile Red staining and machine learning to detect microplastics in bottled drinking water. They found microplastic concentrations up to 152,000 particles per liter, with over 90% of detected particles in the 1-5 micrometer range that falls below current regulatory thresholds. The findings highlight that existing drinking water regulations focusing on particles larger than 20 micrometers may be missing the vast majority of microplastic contamination.
Quantitative image analysis of microplastics in bottled water following Nile Red staining and fluorescence microscopy
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Researchers developed and validated an integrated method for quantifying microplastics in bottled water using Nile Red staining, fluorescent microscopy, and automated image analysis with partial filter interrogation to boost analysis throughput. The method demonstrated high sensitivity for sizing microplastics down to 10 micrometers, with a limit of detection of 1.1 ppb, limit of quantification of 3.4 ppb, linearity between 10 ppb and 1.5 ppm (R2 = 0.99), and repeatability of 11-12% RSD.
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