New protamine quantification method in microtiter plates using o-phthaldialdehyde/N-acetyl-L-cysteine reagent

International Journal of Pharmaceutics
Dirk LochmannA Zimmer

Abstract

Protamine is a well-known excipient in pharmaceutics. It represents a peptide consisting of exclusive aliphatic amino acids, hence it cannot be quantified by UV-spectroscopy (lambdamax 280 nm). A new and sensitive quantification method based on the derivatisation of protamine with ortho-phthaldialdehyde (OPA) in the presents of 2-mercaptoethanol (ME) or N-acetyl-L-cysteine (NAC) in basic aqueous solution using 96-well microtiter plates are introduced in this report. The resulting isoindol derivatives reveal a fluorescence excitation (maximum lambdaex 345 nm) and emission (maximum lambdaem 450 nm) spectra. Derivatives of OPA/NAC reagent were found to be useful for protamine quantification in pharmaceutical nanoparticle preparation containing DNA. A sufficient stability of the isoindol derivatives was shown. It was possible to determine protamine free base, protamine sulphate and protamine chloride with limits of detection less than 1.1 microg/ml.

References

Jan 1, 1976·Nucleic Acids Research·K WehlingK G Wagner
Oct 1, 1985·Analytical Biochemistry·P K SmithD C Klenk
Jun 1, 1971·Analytical Chemistry·M Roth
Jan 1, 1967·Histochemie. Histochemistry. Histochimie·C D NotenboomJ C van de Kamer
Mar 20, 1995·Analytical Biochemistry·J K ChanT A Gill
May 9, 1997·Journal of Chromatography. B, Biomedical Sciences and Applications·B M ChenR Q Zhao
Feb 12, 1998·Gene Therapy·F L SorgiL Huang
Apr 25, 2000·Nucleic Acids Research·M JunghansA Zimmer
Jun 23, 2001·Journal of Chromatography. B, Biomedical Sciences and Applications·M VenzaD Teti
Aug 14, 2001·Journal of Chromatography. B, Biomedical Sciences and Applications·P Campíns-FalcóL A Tortajada Genaro
Aug 23, 2001·Journal of Controlled Release : Official Journal of the Controlled Release Society·M González FerreiroR Bodmeier
Mar 15, 2005·Journal of Microencapsulation·D LochmannA Zimmer
Oct 31, 2008·Talanta·Y Moliner MartinezP Campíns Falcó

❮ Previous
Next ❯

Citations

Feb 10, 2012·Journal of Pharmaceutical and Biomedical Analysis·David Awotwe-OtooRakhi B Shah
Apr 4, 2006·Human Reproduction Update·Rafael Oliva
Feb 2, 2016·Journal of Separation Science·Caroline LamalleMarianne Fillet
Feb 28, 2007·Journal of Biomedical Materials Research. Part a·Kalpna GarkhalNeeraj Kumar
Mar 15, 2011·Journal of Agricultural and Food Chemistry·Roman BuckowYacine Hemar
Jan 25, 2008·The Journal of Physical Chemistry. B·Yun Fei LongYuan Fang Li

❮ Previous
Next ❯

Related Concepts

Related Feeds

Antisense Oligonucleotides: ND

This feed focuses on antisense oligonucleotide therapies such as Inotersen, Nusinursen, and Patisiran, in neurodegenerative diseases including amyotrophic lateral sclerosis.