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Related Experiment Videos

Two methods for quantifying DNA extracted from poly(lactide-co-glycolide) microspheres.

S P Barman1, L Lunsford, P Chambers

  • 1ZYCOS Inc., 44 Hartwell, Lexington, MA 02421, USA.

Journal of Controlled Release : Official Journal of the Controlled Release Society
|December 5, 2000
PubMed
Summary
This summary is machine-generated.

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This study compares UV spectroscopy and PicoGreen dye for quantifying DNA in poly(lactide-co-glycolide) microparticles. PicoGreen offers higher sensitivity but both methods have limitations for encapsulation efficiency and in vitro DNA release studies.

Area of Science:

  • Biomaterials Science
  • Analytical Chemistry
  • Pharmaceutical Sciences

Background:

  • Poly(lactide-co-glycolide) (PLG) microparticles are used for DNA delivery.
  • Accurate quantification of encapsulated DNA is crucial for assessing drug delivery systems.
  • UV spectroscopy and PicoGreen fluorometry are common methods for DNA quantification.

Purpose of the Study:

  • To evaluate the utility and limitations of UV spectroscopy and PicoGreen dye for quantifying DNA in PLG microparticles.
  • To optimize DNA extraction from PLG microspheres for accurate analysis.
  • To assess factors affecting DNA quantification during in vitro release studies.

Main Methods:

  • Comparison of DNA quantification using UV spectroscopy and PicoGreen fluorometry.

Related Experiment Videos

  • Development of a biphasic solvent system for DNA extraction from PLG microspheres.
  • Analysis of DNA encapsulation efficiency and in vitro DNA release.

    • Main Results:

    • PicoGreen demonstrated higher sensitivity (12 pg/ml) than UV spectroscopy (0.5 μg/ml) for DNA detection.
    • Assay conditions for reproducible DNA extraction from PLG microspheres were established.
    • Residual poly(vinyl alcohol) and DNA isoforms (linear, nicked, supercoiled) were found to influence PicoGreen/DNA fluorescence.

    Conclusions:

    • Both UV spectroscopy and PicoGreen fluorometry have limitations for quantifying DNA in PLG microparticles, necessitating careful method selection.
    • Optimized extraction protocols and consideration of interfering substances are vital for accurate DNA quantification in microparticle-based delivery systems.
    • Understanding these limitations is essential for reliable assessment of encapsulation efficiency and in vitro DNA release profiles.