目的:建立同时测定通关藤药材中通关藤苷A、H、I含量的方法.方法:采用超高效液相色谱-串联质谱法,色谱柱为Phenomenex Kinetex XB-C18,流动相为0.1%甲酸溶液-乙腈(梯度洗脱),流速为0.2 mL/min,柱温为40℃,进样量为5 μL,离子源为电喷雾离子源,多反应离子监测模式,正离子扫描,离子源喷射电压为5500 V,雾化气压力为60 psi,加热气压力为60 psi,帘气压力为20 psi,去溶剂温度为600℃.结果:通关藤苷A、H、I检测质量浓度线性范围分别为0.1~10 ng/mL(r=0.9997)、0.025~10 ng/mL(r=0.9995)、0.025~10 ng/mL(r=0.9989);定量限分别为0.1、0.025、0.025 ng/mL,检测险分别为0.05、0.0125、0.0125 ng/mL;精密度、稳定性、重复性试验的RSD均小于4.0%;加样回收率分别为97.67%~99.00%(RSD=0.47%,n=6)、95.00%~101.67%(RSD=2.59%,n=6)、96.67%~103.33%(RSD=2.83%,n=6).结论:该方法操作简便,精密度、稳定性、重复性好,可用于通关藤药材中通关藤苷A、H、I含量的同时测定.%OBJECTIVE:To establish the method for simultaneous determination of tenacissoside A,tenacissoside H and tenacissoside I in Marsdenia tenacissima. METHODS:UPLC-MS/MS method was adopted. The determination was performed on a Phenomenex Kinetex XB-C18column with mobile phase consisted of 0.1% formic acid solution-acetonitrile(gradient elution)at the flow rate of 0.2 mL/min. The column temperature was set at 40 ℃,and sample size was 5 μ L. Multiple reaction monitoring (MRM)mode was adopted with electrospray ion source as ion source,using positive ion scanning. Source jet voltage was 5 500 V,nebulizer pressure was 60 psi,heating pressure was 60 psi,curtain pressure was 20 psi and cone temp was set at 600 ℃. RESULTS:The linear ranges of tenacissoside A,tenacissoside H and tenacissoside I were 0.1-10 ng/mL(r=0.999 7),0.025-10 ng/mL(r=0.999 5),0.025-10 ng/mL(r=0.998 9),respectively;limited of quantation were 0.1,0.025,0.025 ng/mL,limited of detection were 0.05,0.012 5,0.012 5 ng/mL,respectively;RSDs of precision,stability and reproducibility tests were<4.0%. The recoveries were 97.67%-99.00%(RSD=0.47%,n=6),95.00%-101.67%(RSD=2.59%,n=6),96.67%-103.33%(RSD=2.83%, n=6). CONCLUSIONS:The method is simple,precise,stable and reproducible,and can be used for simultaneous determination of tenacissoside A,tenacissoside H and tenacissoside I in M.tenacissima.
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