Simultaneous Determination of 7 Components in Kuhuang Injection by UPLC-MS/MS / 中国药房

ABSTRACT

OBJECTIVE:To establish a method for the content determination of matrine,sophocarpine,emodin,rhein,chloro-genic acid,sarkosaponin a,aloe-emodin in Kuhuang injection.METHODS:UPLC-MS/MS was adopted. The determination was per-formed on Phenomenex Kenetix C18 column with mobile phase consisted of methanol-0.1% formic acid(gradient elution)at the flow rate of 0.5 mL/min. The column temperature was set at 20 ℃,and injector temperature was set at 10 ℃. The equilibrium time was 4 min and sample size was 5 μL. Ionization mode was electrospray ionization with spray voltage of 5500 V and -4500 V,at-omizing air pressure of 4.14×105 Pa,heater pressure of 4.48×105 Pa,curtain air of 1.72×105 Pa and ion source temperature of 600 ℃. The work mode was multiple reaction monitoring mode. RESULTS:The linear range were 1.25-80.0 ng/mL for matrine (r=0.9993),1.10-70.0 ng/mL for sophocarpine(r=0.9995),0.16-10.5 ng/mL for emodin(r=0.9993),2.61-168 ng/mL for rhe-in (r=0.9993),1.50-96.0 ng/mL for chlorogenic acid (r=0.9993),1.48-94.5 ng/mL for sarkosaponin a (r=0.9996) and 6.11-391 ng/mL for aloe-emodin(r=0.9991). The limits of quantification were 0.061,0.109,0.041,1.313,0.500,0.492,3.055 ng/mL,limits of detection were 0.025,0.054,0.016,0.656,0.150,0.148,1.528 ng/mL. RSDs of precision,stability and reproducibility tests were all no more than 3%. The recoveries of them were 95.45%-99.45%(RSD=1.43%,n=6),97.50%-101.00%(RSD=1.50%,n=6),95.67%-101.73%(RSD=2.85%,n=6),97.17%-100.57%(RSD=1.16%,n=6),95.19%-98.90%(RSD=1.71%, n=6),95.38%-103.85%(RSD=3.39%,n=6),95.50%-101.17%(RSD=1.20%,n=6),respectively. CONCLUSIONS:The method is simple,effective,accurate,reliable and suitable for simultaneous determination for 7 components in Kuhuang injection.