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目的:建立同时测定硫磺熏蒸前后当归中阿魏酸、藁本内酯、丁烯基苯酞、洋川芎内酯A、洋川芎内酯H、洋川芎内酯I、欧当归内酯A等7种活性成分含量的方法。方法:采用超高效液相色谱法。色谱柱为Waters BEH C18,流动相为1%醋酸-甲醇(梯度洗脱),柱温为30 ℃,流速为0.35 mL/min,检测波长为254 nm,进样量为2 μL。结果:阿魏酸、藁本内酯、丁烯基苯酞、洋川芎内酯A、洋川芎内酯H、洋川芎内酯I、欧当归内酯A检测进样量的线性范围分别为0.021 6~0.324 0、0.327 2~4.908 0、0.011 51~0.172 7、0.055 0~0.825 0、0.002 248~0.033 72、0.007 2~0.108、 0.001 08~0.016 2 μg(r2=0.999 1~1.000 0);检测限分别为3.7、0.56、0.52、2.02、0.007 6、1.16、0.64 ng;定量限分别为11.5、1.85、1.66、6.26、0.076、3.36、1.79 ng;精密度、稳定性、重复性试验的RSD均<3.0%(n=6);7种成分平均加样回收率为95.79%~101.70%,RSD为2.10%~2.93%(n=6)。硫磺熏蒸后对当归中7种活性成分含量的影响较大,总体呈下降趋势。结论:本研究建立的方法的精密度、稳定性、重复性均较好,可用于硫磺熏蒸前后当归中7种活性成分的含量测定;硫磺熏蒸后会降低当归中活性成分的含量。
OBJECTIVE: To establish the method for content determination of 7 kinds of active components that as ferulic acid, ligustilide, n-butylidenephthalide, senkyunolide A, senkyunolide H, senkyunolide I and levistilide A in Angelica sinensis before and after sulfur fumigation simultaneously. METHODS: UPLC method was adopted. The determination was performed on Waters BEH C18 column with mobile phase consisted of 1%acetic acid-methanol (gradient elution) at flow rate of 0.35 mL/min. The column temperature was 30 ℃, and detection wavelength was set at 254 nm. The sample size was 2 μL. RESULTS: The linear range for ferulic acid, ligustilide, n-butylidenephthalide, senkyunolide A, senkyunolide H, senkyunolide I and levistilide A was 0.021 6-0.324 0, 0.327 2-4.908 0, 0.011 51-0.172 7, 0.055 0-0.825 0, 0.002 248-0.033 72, 0.007 2-0.108, 0.001 08-0.016 2 μg (r2= 0.999 1-1.000 0), respectively. The limits of detection were 3.7, 0.56, 0.52, 2.02, 0.007 6, 1.16, 0.64 ng, respectively. The limits of quantitation were 11.5, 1.85, 1.66, 6.26, 0.076, 3.36, 1.79 ng, respectively. RSDs of precision, stability and reproducibility tests were lower than 3.0% (n=6). The average recovery rates of 7 kinds of active components were from 95.79% to 101.70% (RSD=2.10%-2.93%, n=6). The contents of active 7 kinds of active components in A. sinensis were greatly influenced by sulfur fumigation and had a downward trend in general. CONCLUSIONS: The established method is simple with good precision,stability and reproducibility and can be used for the content determination of 7 kinds of active components in A. sinensis before and after sulfur fumigation. The sulfur fumigation will decrease the contents of active components in A. sinensis.
当归硫磺熏蒸超高效液相色谱法含量测定
Angelica sinensisSulfur fumigationUPLCContent determination
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