[Nano ultra high performance liquid chromatography coupled with time-of-flight mass spectrometer for detecting low molecular weight polypeptides in zebrafish embryos].
OBJECTIVE
To apply the nano-ultra high performance liquid chromatography coupling with time-of-flight mass spectrometer (nano-UPLC-TOF-MS) to detect the low molecular weight polypeptides in zebrafish embryos.


METHODS
Zebrafish embryos were collected 24 hours after fertilization and the outer chorion was removed. And then the yolk was removed with mechanical forces, of which the efficiency was validated by microscope and SDS-PAGE; the low molecular weight polypeptide was extracted with corresponding solution, and followed by separation and detection with nano-UPLC-TOF-MS.


RESULTS
Mechanical vortex almost completely removed yolk cells of zebrafish embryos, from which the special solution extracted effectively low molecular weight polypeptides for the later detection. As demonstrated by nano-UPLC-TOF-MS, the polypeptide peak showed between 22 and 24. 5 min, and the m/z ration of most of polypeptides were between 800-1400.


CONCLUSIONS
Nano-OPLC-TOF-MS is effective to detect low molecular weight polypeptides, and low molecular weight peptides definitely exist in zebrafish embryos, of which the relative molecular mass seems to be 800 to 1400, probably consists of about 10 residues.
Yang
Han-shuo
HS
State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
Tang
Ming-hai
MH
Deng
Hong-xin
HX
Yang
Jin-liang
JL
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D010455:Peptides / Q000032:analysis*; D015027:Zebrafish / Q000196:embryology*
D010455:Peptides