TY - JOUR
T1 - Analysis of adenosine phosphates in HepG-2 cell by a HPLC-ESI-MS system with porous graphitic carbon as stationary phase
AU - Wang, Jun
AU - Lin, Tao
AU - Lai, Jiaping
AU - CAI, Zongwei
AU - Yang, M. S.
N1 - Funding Information:
The financial supports of the Health and Health Services Research Fund from Food and Health Bureau of Hong Kong (05060141) on this study are acknowledged. T. Lin was supported by the special post-graduate studentship from the Research Grant Council (2005–2007) of Hong Kong.
PY - 2009/7/15
Y1 - 2009/7/15
N2 - A high performance liquid chromatography coupled with electrospray ionization/mass spectrometry method was developed for the determination of adenosine 5′-monophosphate (AMP), adenosine 5′-diphosphate (ADP), and adenosine 5′-triphosphate (ATP) in the extract of HepG-2 cells. The chromatographic conditions were optimized by using porous graphitic carbon as the stationary phase for the retention and separation of the AMP, ADP and ATP. Negative-ion mode ESI-MS in basic mobile phase was applied to improve the method sensitivity. An external calibration method with linear ranges from 0.22 to 57.80 μM for AMP, from 0.59 to 117.37 μM for ADP, and from 0.49 to 98.81 μM for ATP was used for quantitative analysis. The levels of ATP, ADP, and AMP in HepG-2 cells treated with benzo[a]pyrene with different time periods were determined. Total adenine nucleotides and the energy charge potential were calculated for the investigation of the effect of benzo[a]pyrene on cell energy metabolism.
AB - A high performance liquid chromatography coupled with electrospray ionization/mass spectrometry method was developed for the determination of adenosine 5′-monophosphate (AMP), adenosine 5′-diphosphate (ADP), and adenosine 5′-triphosphate (ATP) in the extract of HepG-2 cells. The chromatographic conditions were optimized by using porous graphitic carbon as the stationary phase for the retention and separation of the AMP, ADP and ATP. Negative-ion mode ESI-MS in basic mobile phase was applied to improve the method sensitivity. An external calibration method with linear ranges from 0.22 to 57.80 μM for AMP, from 0.59 to 117.37 μM for ADP, and from 0.49 to 98.81 μM for ATP was used for quantitative analysis. The levels of ATP, ADP, and AMP in HepG-2 cells treated with benzo[a]pyrene with different time periods were determined. Total adenine nucleotides and the energy charge potential were calculated for the investigation of the effect of benzo[a]pyrene on cell energy metabolism.
KW - Adenosine nucleotides
KW - Benzo[a]pyrene
KW - HepG-2 cell
KW - LC-ESI-MS
KW - Porous graphitic carbon
UR - http://www.scopus.com/inward/record.url?scp=67649236203&partnerID=8YFLogxK
U2 - 10.1016/j.jchromb.2009.05.027
DO - 10.1016/j.jchromb.2009.05.027
M3 - Journal article
C2 - 19502115
AN - SCOPUS:67649236203
SN - 1570-0232
VL - 877
SP - 2019
EP - 2024
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
IS - 22
ER -