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Analysis of single nucleotide polymorphism sites in exon 4 of the p53 gene using
high-performance liquid chromatography electrospray ionization mass spectrometry tandem mass spectrometry Renfang Song,a,b Wenbing
Zhang,a Huayong Chen,a,b Huimin Ma,a,b Yulian Dong,a Guoying Sheng,a Zhen Zhoua and Jiamo
Fua,* aState Key Laboratory of Organic Geochemistry, Guangzhou Research Center of Mass Spectrometry, Guangzhou Institute of
Geochemistry, Chinese Academy of Sciences, Guangzhou 510640, China. E-mail: fujm@gig.ac.cn bGraduate School of the Chinese Academy of Sciences,
Beijing, China
ABSTRACT:
Three groups of four oligonucleotides with special single nucleotide polymorphisms (SNP) sites in exon 4 of the p53 gene were analyzed with ion-pair reversed-
phase high-performance liquid chromatography electrospray ionization mass spectrometry tandem mass spectrometry. The retention order of four oligonucleotides with SNPs was
C < G < A < T, regardless of whether the polymorphisms were at the 3′ end, the 5′ end, or the middle of the oligonucleotides. The charge state of the
molecular ion affects the MS/MS spectra of the oligonucleotides. SNPs at the 3′ end can be easily identified from the fragmentation pattern of the 2- charge state, but not
from the 3- charge state, especially from the w1 fragment. The single base may be taken as the symbol of the 5′ end SNP site derived from
[M3H]2, but not from the [M3H]2 charge state. The oligonucleotides with SNPs in the middle were also determined from
the [M2H]2 precursor ion.
Keywords:
single nucleotide polymorphisms (SNPs); high-performance liquid chromatography; electrospray ionization mass
spectrometry; SNP site; retention order; charge state; product ion
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