Qualitative analysis of bis-(3'-5')-cyclic dimeric adenosine monophosphate of Porphyromonas gingivalis by high performance liquid chromatography coupled with mass spectrometry.
- Author:
Yongmei TAN
;
Xiaojun YANG
;
Juan DU
;
Wanghong ZHAO
;
Xiaodan CHEN
;
Jin HOU
- Publication Type:Journal Article
- MeSH: Chromatography, High Pressure Liquid; Cyclic AMP; chemistry; metabolism; Periodontitis; Porphyromonas gingivalis; metabolism; Tandem Mass Spectrometry
- From: West China Journal of Stomatology 2016;34(3):307-311
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo test whether Porphyromonas gingivalis (P. gingivalis) could produce bacterial signal molecule, bis-(3'-5')-cyclic dimeric adenosine monophosphate (c-di-AMP) and lay the foundation for explorations of its roles in life metabolism and periodontitis immunity of P. gingivalis.
METHODSP. gingivalis standard strain ATCC33277 was used as the experimental strain to extract nucleic acids from the bacteria. Then, c-di-AMP was detected using high performance liquid chromatography coupled with mass spectrometry (HPLC-MS/MS). Subsequently, HPLC was used to validate the sample further.
RESULTSBased on the signal/noise (S/N) for 3 : 1, the limit of determination of HPLC-MS/MS for peak time of c-di-AMP standard substances was 7.49 min and nucleic acid extractions from P. gingivalis was 8.82 min (S/N > 3). Further confirmation of HPLC showed that nucleic acid extractions from both P. gingivalis and c-di-AMP standard substances pre- sented goal absorbent peaks at 15.7 min, with the same ultraviolet absorbent spectrum.
CONCLUSIONThe nucleic acid extrac- tions from P. gingivalis contained c-di-AMP, which shows that P. gingivalis could produce c-di-AMP.