A sensitive HPLC method for the determination of fatty acids using 2-(2-(pyren-1-yl)-1Hbenzo[
d]imidazol-1-yl) ethyl-p-toluenesulfonate (PBIOTs) as a novel labeling reagent with fluorescence
detection has been developed. PBIOTs could easily and quickly label fatty acids in the presence
of the K2CO3 catalyst at 90C for 30 min in N,N-dimethylformamide solvent. Fatty acids derivatives
were separated on a reversed phase Eclipse XDB-C8 column by HPLC in conjunction with gradient
elution. The corresponding derivatives were identified by post-column APCI-MS in positive ion
detection mode. PBIOTs-fatty acid derivatives gave an intense molecular ion peak at m=z
[MþH]þ; with MS=MS analysis, the collision induced dissociation spectra of m=z [MþH]þ
produced the specific fragment ions at m=z [MþH–319]þ and m=z 319.0 (here, m=z 319 is the
core structural moiety of the PBIOTs molecule). The fluorescence excitation and emission wavelengths
of the derivatives were kex¼350 nm and kem¼402 nm, respectively. Linear correlation coefficients
for all fatty acid derivatives were 0.9985. Detection limits for the labeled fatty acids, at a
signal-to-noise ratio of 3:1, were in the range of 10.32– 44.28 fmol. From the extracts of Stellera
chamaejasma L, free fatty acids were sensitively determined.
d]imidazol-1-yl) ethyl-p-toluenesulfonate (PBIOTs) as a novel labeling reagent with fluorescence
detection has been developed. PBIOTs could easily and quickly label fatty acids in the presence
of the K2CO3 catalyst at 90C for 30 min in N,N-dimethylformamide solvent. Fatty acids derivatives
were separated on a reversed phase Eclipse XDB-C8 column by HPLC in conjunction with gradient
elution. The corresponding derivatives were identified by post-column APCI-MS in positive ion
detection mode. PBIOTs-fatty acid derivatives gave an intense molecular ion peak at m=z
[MþH]þ; with MS=MS analysis, the collision induced dissociation spectra of m=z [MþH]þ
produced the specific fragment ions at m=z [MþH–319]þ and m=z 319.0 (here, m=z 319 is the
core structural moiety of the PBIOTs molecule). The fluorescence excitation and emission wavelengths
of the derivatives were kex¼350 nm and kem¼402 nm, respectively. Linear correlation coefficients
for all fatty acid derivatives were 0.9985. Detection limits for the labeled fatty acids, at a
signal-to-noise ratio of 3:1, were in the range of 10.32– 44.28 fmol. From the extracts of Stellera
chamaejasma L, free fatty acids were sensitively determined.
The research has been published on tJournal of Liquid Chromatography & Related Technologies, 33:859–874, 2010
Copyright # Taylor & Francis Group, LLC
ISSN: 1082-6076 print/1520-572X online
DOI: 10.1080/10826071003684620
Copyright # Taylor & Francis Group, LLC
ISSN: 1082-6076 print/1520-572X online
DOI: 10.1080/10826071003684620
Cuihua Song,1 Zhiwei Sun,2,3 Lian Xia,1,2,3 Yourui Suo,2 and
Jinmao You1,2
Jinmao You1,2