Formic acid hydrolysis/liquid chromatography isotope dilution mass spectrometry: An accurate method for large DNA quantification.

Journal of chromatography. A

PubMedID: 27665496

Shibayama S, Fujii SI, Inagaki K, Yamazaki T, Takatsu A. Formic acid hydrolysis/liquid chromatography isotope dilution mass spectrometry: An accurate method for large DNA quantification. J Chromatogr A. 2016;.
Liquid chromatography-isotope dilution mass spectrometry (LC-IDMS) with formic acid hydrolysis was established for the accurate quantification of ?DNA. The over-decomposition of nucleobases in formic acid hydrolysis was restricted by optimizing the reaction temperature and the reaction time, and accurately corrected by using deoxynucleotides (dNMPs) and isotope-labeled dNMPs as the calibrator and the internal standard, respectively. The present method could quantify ?DNA with an expanded uncertainty of 4. 6% using 10fmol of ?DNA. The analytical results obtained with the present method were validated by comparing with the results of phosphate-base quantification by inductively coupled plasma-mass spectrometry (ICP-MS). THE RESULTS
showed good agreement with each other.We conclude that the formic acid hydrolysis/LC-IDMS method can quantify ?DNA accurately and is promising as the primary method for the certification of DNA as reference material.