XU He1,2, JIN Shi-ying1,2, XU Ping-hua1, YUAN Hai-long1*, JIN Shi-xiao1,2, MENG Xiao-lin3, HAN Jin1*
1 Department of Pharmacy,302 Military Hospital of PLA,Beijing 100039,China; 2 School of Pharmaceutical Sciences,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China; 3 State Key Laboratory of Virology,Wuhan University,Wuhan 430072,China
Abstract��
Objective To establish a method for determination of cecropin from Plutella xylostella (pxCECA1) by RP-HPLC. METHODS An Agilent Alltima C18 column (4.6 mm×250 mm,5 μm) was adopted,and the mobile phase was composed of methanol-acetonitrile-0.05% TFA (2.5:2.5:95) at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 275 nm and the column temperature was 25 ��. RESULTS The linear range of the calibration curve for cecropin was 0.08-0.800 μg (r=0.999 4,n=6). The average recovery was 99.69%,and RSD was 1.20% (n=6),respectively. CONCLUSION The method is rapid,simple,and accurate with good reproducibility. It can be used for quantitative analysis and quality control of cecropin from Plutella xylostella.
WANG H,MENG X L,XU J P,et al. Production,purification,and characterization of the cecropin from Plutella xylostella,pxCECA1,using an intein-induced self-cleavable system in Escherichia coli. Appl Microbiol Biotechnol,2012,94(4): 1031-1039.
[4]
HU Q,LIU Y D. Progress and summarization of study on diamondback moth (Plutella xylostella L.) . J Tianjin Agricul Coll�����ũѧԺѧ����,2003,10(1): 33-36.
[5]
SUN F. HPLC Purification of Antimicrobial Peptides from Northeast Forest Frogs and the Study of Anti-Influenza Virus Effect in Vitro . Harbin: Northeast Forestry University,2007.
[6]
LORAINE S,JEFFREY N W,PAUL H A. Antibacterial and antimembrane activities of cecropin A in Escherichia coli. Antimicrob Agents Chemother,2000,42(3): 602-607.
[7]
ALISON J,MOORE A,PAUL M,et al.Extraction of the synthetic lytic peptide,cecropin B,frombiological fluid and analysis using reversed-phase high performance liquid chromatography. J Chromatog B,1995,672(2): 225-231.
[8]
YANG Z H,YANG X Y,YUAN X H,et al. Purification of the antimicrobial peptide AMP-1 of cyclopelta parva distant by HPLC . J Southwest Agricultural Univ(����ũҵ��ѧѧ��),2001,23(2): 104-107.
[9]
GONG X,SHI Y H,LE G W. Purification of the antimicrobial peptide MDL-1 from Musca domestica tarvae and its effect on Escherichia coli ultrastucture . Acta Entomol Sin(����ѧ��),2004,47(1): 8-13.
[10]
SUN D P,HU Y Q. Stabilizing excipients in the freeze-dried protein formulations and their protective mechanisms . Prog Pharm Sci (ҩѧ��չ),2003,27(4): 201-205.
2013, Vol. 48 
