OBJECTIVETo establish the method for the quality control of Xuefuzhuyu granule.

METHODThe content of Semen Armeniacae Amarum PE, Hydroxysafflor yellow A, paeoniflorin and ferulic acid in Xuefuzhuyu granule were determined by HPLC under multi-wavelength of 210, 403, 230, 316 nm.

RESULTThe calibration curve of Semen Armeniacae Amarum PE. hydroxysafflor yellow A, paeoniflorin and ferulic acid were linear within 0.1154-0.9232 (r = 0.9998), 0.0219-0.1754 (r = 0.9994), 0.3952-3.1616 (r = 0.9995), and 0.0426-0.3408 microg (r = 0.9998), respectively. The average recoveries were 101.78% (RSD was 1.7%), 99.60% (RSD was 2.9%), 98.90% (RSD was 2.0%), and 100.31% (RSD was 1.8%), respectively.

CONCLUSIONThe method of quantification is accurate, rapid and reliable for the quality control of Xuefuzhuyu granule.

Benzoates ; analysis ; Bridged-Ring Compounds ; analysis ; Chalcone ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; analysis ; Drugs, Chinese Herbal ; analysis ; chemistry ; Glucosides ; analysis ; Monoterpenes ; Quinones ; analysis ; Reproducibility of Results

In order to mine candidate vaccine antigens against ticks and to control ticks safely and effectively,the aim of this study was to immunize rabbits with purified aquaporins(AQPs)rD-mAQP1,rDmAQP2 and rDmAQP3 of Dermacentor marginatus.Blood collections for Western blot and ELISA tests were performed.The anti-tick challenge was conducted.The optimal expression conditions of rDmAQP1,rDmAQP2 and rDmAQP3 were screened by SDS-PAGE gel electrophore-sis.The three recombinant proteins were purified by HisSepNi-NTA6FF purification column.Rab-bits were divided into four groups of three rabbits each,including a control group and three immu-nized groups.The three purified recombinant proteins were separately immunized to three groups of rabbits,and the rabbits were immunized once on the 0th,14th and 21st day.Blood samples were collected every 7 days to prepare polyclonal antibodies.The reactivity was detected by Western blot and the antibody titer was detected by ELISA.Tick challenge test was carried out after the anti-body titer increased.The results showed that the optimal expression conditions for rDmAQP1 were induced for 8 h at IPTG concentration of 1.0 mmol/L and 37 ℃;the optimal expression conditions for rDmAQP2 were induced for 7 h at IPTG concentration of 1.0 mmol/L and 37 ℃;and the opti-mal expression conditions for rDmAQP3 were induced for 5 h at IPTG concentration of 1.0 mmol/L and 37 ℃.Western blot results showed that rDmAQP1,rDmAQP2 and rDmAQP3 all had certain reactivity.The ELISA results showed that the antibody titers of rabbits immunized with rD-mAQP1,rDmAQP2 and rDmAQP3 were as follows:the total anti-tick effect of rDmAQP1 protein was 79.74％,and the inhibition rates on average full-blooded tick weight,average egg weight and average egg hatching rate were 9.43％,25.17％and 63.81％,respectively.The total anti-tick effect of rDmAQP2 protein was 78.78％,and the inhibition rates of average full-blooded tick weight,av-erage egg weight and average egg hatching rate of Dermacentor marginatus were 8.30％,20.14％and 68.26％,respectively.The total anti-tick effect of rDmAQP3 protein was 87.91％,and the inhi-bition rates of average full-blooded tick weight,average egg weight and average egg hatching rate were 3.23％,22.47％and 80.5％,respectively.Through serological test and anti-tick test,it has been found that rDmAQP1,rDmAQP2 and rDmAQP3 all have the potential of candidate antigens against ticks,among which rDmAQP3 has the best immune effect,which lays a foundation for the study of the function of rDmAQP1,rDmAQP2 and rDmAQP3.