Quantification of unknown samples using standard curves

Importance of the Topic

Standard curves are fundamental for quantifying analyte concentrations in unknown samples by correlating absorbance to known standards. They are widely used across biochemical assays, environmental analysis, and quality control workflows. Implementing robust calibration models enhances accuracy and reproducibility of measurements.

Objectives and Study Overview

This application note demonstrates the use of the Lunatic and Little Lunatic platforms to generate, save, and apply standard curves for sample quantification. It highlights user workflows for creating new curves or reusing saved ones and compares single-wavelength measurements versus dual-wavelength difference methods.

Methodology and Instrumentation

A region of lowest absorbance is used for automatic background correction, eliminating the need for blank runs. Users can define reference concentrations, select curve types proportional linear point-to-point quadratic 4-parameter and set wavelength(s). The Lunatic platform supports flexible report customization including HTML XML TXT CSV XLSX and PDF. Little Lunatic uses predefined templates and generates HTML XML TXT and CSV.

Results and Discussion

Measured standard points are plotted and fitted to the chosen regression model. The regression formula provides concentration estimates for unknown samples and an R2 value indicates fit quality. The dual-wavelength approach improves precision by reducing standard deviation of results as demonstrated by comparative data.

Benefits and Practical Applications

• Enhanced precision using two-wavelength corrections
• Automated background subtraction simplifies workflow
• Multiple curve models accommodate diverse assay behaviors
• Flexible reporting supports data export and compliance
• Rapid quantification for high-throughput environments

Future Trends and Potential Applications

Ongoing advances in spectral processing and machine learning may enable adaptive curve fitting and real-time quality checks. Integration with laboratory information management systems could streamline data management. Expansion to multiplex assays and novel detector technologies will further broaden the methods applicability.

Conclusion

The Lunatic standard curve application offers a comprehensive solution for accurate concentration determination. Its flexible calibration options automated correction routines and robust reporting features make it suitable for research QAQC and industrial analytics.

Reference

Unchained Labs Application Note Quantification of unknown samples using standard curves Rev B 2020.