The article reviews the application of next-generation sequencing (NGS) in dietary studies, focusing on its potential and limitations. It highlights the importance of accurate dietary data for understanding food web dynamics and ecosystem functions. Traditional methods, such as visual observations and molecular approaches, have limitations in scale and resolution. NGS has revolutionized dietary studies by enabling the simultaneous analysis of many species, reducing costs, and increasing data volume. The article discusses key considerations for designing NGS diet assessments, including the choice of DNA barcodes, amplification efficiency, reference databases, and contamination prevention. It also addresses technical aspects such as experimental design, data analysis, and validation of data accuracy. The review emphasizes the need for careful validation to ensure the reliability of NGS-based dietary data, particularly in quantifying food intake and avoiding biases.The article reviews the application of next-generation sequencing (NGS) in dietary studies, focusing on its potential and limitations. It highlights the importance of accurate dietary data for understanding food web dynamics and ecosystem functions. Traditional methods, such as visual observations and molecular approaches, have limitations in scale and resolution. NGS has revolutionized dietary studies by enabling the simultaneous analysis of many species, reducing costs, and increasing data volume. The article discusses key considerations for designing NGS diet assessments, including the choice of DNA barcodes, amplification efficiency, reference databases, and contamination prevention. It also addresses technical aspects such as experimental design, data analysis, and validation of data accuracy. The review emphasizes the need for careful validation to ensure the reliability of NGS-based dietary data, particularly in quantifying food intake and avoiding biases.