HPLC-ESI-MS法同时测定印度獐牙菜中5种酮类化合物的含量

Simultaneous Determination of Five Xanthones in Swertia chirayita by HPLC-ESI-MS

目的：建立同时测定印度獐牙菜中5种酮类化合物含量的方法。方法：采用高效液相色谱-电喷雾-离子阱质谱（HPLC-ESI-MS）法。色谱条件：色谱柱为Waters Symmetry-C18，流动相为水（5 mmol/L乙酸铵-0.1％甲酸）-甲醇（梯度洗脱），流速为0.8 ml/min，检测波长为254 nm，柱温为30℃，进样量为10μl；质谱条件：正离子检测，离子源喷射电压为4.5 kV，毛细管温度为200℃，毛细管电压为25 V，鞘气流速为35 arb，辅助气流速为20 arb，碰撞气为氦气，检测方式为多级全扫描质谱。结果：1-羟基-3，5-二甲氧基酮、1-羟基-3，4，5-三甲氧基酮、1，8-二羟基-3，5-二甲氧基酮、1-羟基-2，3，4，5-四甲氧基酮、1-羟基-2，3，4，7-四甲氧基酮检测质量浓度线性范围均为0.5～15.00μg/ml（r均大于0.9990）；精密度、稳定性、重复性试验的RSD＜2％；加样回收率分别为100.03％～103.59（RSD＝0.99％，n＝9）、97.41％～100.89％（RSD＝1.15％，n＝9）、99.13％～101.68％（RSD＝1.40％，n＝9）、100.21％～103.51％（RSD＝1.33％，n＝9）、100.56％～103.92％（RSD＝1.37％，n＝9）。结论：该方法操作简便，稳定性和重复性好，可用于印度獐牙菜中5种酮类化合物含量的同时测定。

OBJECTIVE：To establish a method for the simultaneous determination of 5 xanthones in Swertia chirayita. METH-ODS：HPLC-EST-MS was conducted. The chromatographic conditions：the column was Waters Symmetry-C18 with mobile phase of water （5 mmol/L ammonium acetate- 0.1% formic acid）-methanol （gradient elution） at a flow rate of 0.8 ml/min,the detection warelength was 254 nm,column temperature was 30 ℃,and the injection volume was 10 μl;MS conditions：it was detected by positive ion,ion source ejection voltage was 4.5 kV,capillary temperature was 200 ℃,capillary voltage was 25 V,sheath gas flow rate was 35 arb,auxiliary gas flow rate was 20 arb,collision gas was helium,and detection method was multi-stage full scan mass spectrometry. RESULTS：The linear ranges for 1-hydroxy-3,5-dimethoxyxanthone,1-hydroxy-3,4,5-trimethoxyxanthone,1, 8-dihydroxy-3,dimethoxyxanthone,1-hydroxy-2,3,4,5-tetramethoxyxanthone and 1-hydroxy-2,3,4,7-tetramethoxyxanthone were all 0.5-15.00 μg/ml（r〉0.999 0）;RSDs of precision,stability and reproducibility tests were lower than 2%;recoveries were 100.03%-103.59%（RSD=0.99%,n=9）for 1-hydroxy-3,5-dimethoxyxanthone,97.41%-100.89%（RSD=1.15%,n=9）for 1-hy-droxy-3, 4, 5-trimethoxyxanthone, 99.13% -101.68%（RSD=1.40% , n=9） for 1, 8-dihydroxy-3, 5-dimethoxyxanthone, 100.21%-103.51%（RSD=1.33%,n=9）for 1-hydroxy-2,3,4,5-tetramethoxyxanthone,100.56%-103.92%（RSD=1.37%,n=9） for 1-hydroxy-2,3,4,7-tetramethoxyxanthone. CONCLUSIONS：The method is simple,stable and reproducible,and can be used for the simultaneous determination of 5 xanthones in S. chirayita.