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Absorbance Spectroscopy: Spectral Artifacts and Other Sources of Error

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Encyclopedia of Biophysics
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Synonyms

Deviations from Beer–Lambert Law; Light scattering; Solvent absorption; Spectrophotometric error; Stray light

Definition

Modern spectrophotometers and data acquisition software are usually reliable and easy to use, but the spectra that they produce must always be regarded critically, as there are many possible sources of error and measurement uncertainty. Some of these relate to sample purity and identity. Others relate to the quality and calibration of the instrument, experimental technique, choice of sample cells and wavelength ranges, correct use of software for data acquisition and analysis, appropriate scan parameters that optimize signal-to-noise ratios yet ensure the fidelity of the recorded spectrum, and uncertainties associated with data analysis (e.g., the use of standard curves). A good understanding of spectroscopic principles, and proper instruction and supervision in the use of spectroscopic instrumentation, is essential for reliable spectroscopic work. Other...

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References

  • Gordon DJ, Holzwarth G (1971) Artifacts in the measured optical activity of membrane suspensions. Arch Biochem Biophys 142:481–488

    Article  CAS  Google Scholar 

  • van de Hulst HC (1981) Light scattering by small particles. Dover Publications Inc., New York

    Google Scholar 

  • Kelly SM, Jess TJ, Price NC (2005) How to study proteins by circular dichroism. Biochim Biophys Acta 1751:119–139

    Article  CAS  Google Scholar 

  • Mao D, Wallace BA (1984) Differential light scattering and absorption flattening optical effects are minimal in the circular dichroism spectra of small unilamellar vesicles. Biochemistry 23:2667–2673

    Article  CAS  Google Scholar 

  • Meeten GH (1981) Conservative dichroism in the Rayleigh-Gans-Debye approximation. J Colloid Interface Sci 84:235–239

    Google Scholar 

  • Nordh J, Deinum J, Nordén B (1986) Flow orientation of brain microtubules studied by linear dichroism. Eur Biophys J 14:113–122

    Article  CAS  Google Scholar 

  • Waldron DE, Marrington R, Grant MC, Hicks MR, Rodger A (2010) Capillary circular dichroism. Chirality 22:E136–E141

    Article  CAS  Google Scholar 

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Correspondence to Paul Wormell .

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© 2018 European Biophysical Societies' Association (EBSA)

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Wormell, P., Rodger, A. (2018). Absorbance Spectroscopy: Spectral Artifacts and Other Sources of Error. In: Roberts, G., Watts, A. (eds) Encyclopedia of Biophysics. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-35943-9_776-1

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  • DOI: https://doi.org/10.1007/978-3-642-35943-9_776-1

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  • Print ISBN: 978-3-642-35943-9

  • Online ISBN: 978-3-642-35943-9

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