Advantages of nanodrop over traditional spectrophotometr

 Advantages of nanodrop over traditional spectrophotometry.

Author: Akash Saha, Student, NFSU-Tripura Campus.

1. Introduction

Brief overview of spectrophotometry:

Spectrophotometry refers to the process of measuring how materials engage with light, including their reflection, transmission, scattering, absorption, and light emission. This technique requires accurate definitions of various quantities to ensure precise measurements and reliable interpretations of optical characteristics.

Introduction to Nanodrop technology:

Nanodrop spectrophotometers use microvolume samples (around 2 μL) and fiber optic technology to measure nucleic acids, proteins, and biomolecules, minimizing sample waste compared to traditional methods.

2. Traditional Spectrophotometry: Key Features

Basic principles of traditional spectrophotometry:

A spectrophotometer operates on the principle that a compound absorbs or transmits light at specific wavelengths. It obeys Beer's Law. 

Working of Spectrophotometer:

A spectrophotometer typically combines two components: a spectrometer, which generates, disperses, and measures light, and a photometer, which uses a photoelectric detector to measure light intensity. 

Spectrometer: It generates a specific range of light wavelengths. A collimator directs a  straight beam of light through a monochromator, which splits it into different wavelengths. A wavelength selector then filters and transmits only the required wavelengths, as depicted in Figure 1.

Photometer: When light of a specific wavelength passes through the sample solution in a cuvette, the photometer measures the absorbed photons and transmits a signal to a galvanometer or digital display, as shown.

3. Nanodrop Technology: A Modern Alternative.

Basic principles of Nanodrop technology: 

Nanodrop spectrophotometers operate using UV-Vis absorbance, detecting characteristic peaks for various biomolecules.

Working of Nanodrop technology

Figure 2: Techniques of nanodrop (Source: ResearchGate)

4. Nanodrop Vs Traditional Spectrophotometry:

5. Applications and Use Cases:

I. Nucleic acid quantification[10]

II. Contaminant identification

III. Nucleic acid quantification in RT-qPCR workflows

IV. Protein quantification

V. Choosing the best quantification assay for your protein or peptide sample

VI. Quality control laboratories

VII. Compliance in pharmaceuticals, biotechnology, and other regulated environments

VIII. Bacterial culture growth (OD600)

IX. Time-based kinetic measurements [11]

6. References

[1] T. A. Germer, J. C. Zwinkels, and B. K. Tsai, “Theoretical Concepts in Spectrophotometric Measurements,” Experimental Methods in the Physical Sciences, vol. 46, pp. 11–66, Jan. 2014, doi: 10.1016/B978-0-12-386022-4.00002-9.

[2] P. Desjardins and D. Conklin, “NanoDrop Microvolume Quantitation of Nucleic Acids,”J Vis Exp, no. 45, 2010, doi: 10.3791/2565.

[3] J. H. Hardesty, B. Attili, and C. College, “Spectrophotometry and the Beer-Lambert Law: An Important Analytical Technique in Chemistry,” 2010.

[4] “2.1.5: Spectrophotometry - Chemistry LibreTexts.” Accessed: Sep. 07, 2024. [Online].

Available:

https://chem.libretexts.org/Bookshelves/Physical_and_Theoretical_Chemistry_Textbook_Maps/Supplemental_Modules_(Physical_and_Theoretical_Chemistry)/Kinetics/02%3A_Reaction_Rates/2.01%3A_Experimental_Determination_of_Kinetics/2.1.05%3A_Spectrophotometry

[5] “NanoDrop Spectrophotometer Resources | Thermo Fisher Scientific - IN.” Available: https://www.thermofisher.com/in/en/home/industrial/spectroscopy-elemental-isotopeanalysis/molecular-spectroscopy/uv-visspectrophotometry/instruments/nanodrop/resources.html

[6] P. Desjardins, J. B. Hansen, and M. Allen, “Microvolume protein concentration determination using the NanoDrop 2000c Spectrophotometer,” Journal of Visualized Experiments, no. 33, 2010, doi: 10.3791/1610.

[7] S. K. Gupta, K. Tapadia, and A. Sharma, “A sensitive nanodrop method for the microlevel determination of cationic surfactant methyltrioctylammonium chloride (MTOAC) in biological fluids and environmental samples,” Journal of the Iranian Chemical society, vol. 18, no. 2, pp. 343–349, Feb. 2021, doi: 10.1007/S13738-020-02029-3.

[8] S. J. Bunu, D. Otele, T. Alade, and R. Dodoru, “Determination of serum DNA purity among patients undergoing antiretroviral therapy using NanoDrop-1000 spectrophotometer and polymerase chain reaction,” Biomedical and Biotechnology Research Journal, vol. 4, no. 3, pp. 214–219, Jul. 2020, doi:10.4103/BBRJ.BBRJ_68_20.

[9] Y. Vicente-Martínez, M. Caravaca, and M. Briceño, “Spectrophotometric nanodrop system for quantification of trace concentrations of ibuprofen in water samples using functional magnetic nanoparticles,” Microchemical Journal, vol. 180, Sep.2022, doi: 10.1016/J.MICROC.2022.107555.

[10] P. Desjardins and D. Conklin, “NanoDrop Microvolume Quantitation of Nucleic Acids,”J Vis Exp, no. 45, 2010, doi: 10.3791/2565.

[11] “NanoDrop Microvolume Spectrophotometer Applications | Thermo Fisher Scientific -IN.”Available:

https://www.thermofisher.com/in/en/home/industrial/spectroscopy-elemental-isotopeanalysis/molecular-spectroscopy/uv-visspectrophotometry/instruments/nanodrop/applications.html