神香草水提物的主成分分析及含量测定

Analysis of Main Components of Hyssopus officinalis Water Extract and Content Determination

目的：研究神香草水提物中的主要成分,并测定其含量。方法：采用液相色谱-质谱联用技术分析神香草水提物中的主要成分。色谱柱为Waters 5 C18-DAQ,流动相为0.2%乙酸水溶液-乙腈（梯度洗脱）,流速为1.0 ml/min,柱温为30℃,进样量为10μl,检测波长为360 nm;离子源为电喷雾（ESI）离子源,采用负离子扫描模式,离子源温度为400℃,质量扫描范围为100～1 000 Da。选择迷迭香酸为指标成分,并采用反相高效液相色谱法对其含量进行测定。色谱柱为Phenomenex C18,流动相为0.2%甲酸水溶液-乙腈（75∶25,V/V）,检测波长为330 nm,流速为1.0 ml/min,柱温为30℃,进样量为10μl。结果：主成分（22.65 min）的离子质量数为359.2,推测其为迷迭香酸。迷迭香酸的质量浓度在4.52～22.60μg/ml范围内与峰面积呈良好线性关系（r=0.999 9）;精密度、稳定性、重复性试验的RSD〈2%;平均加样回收率为99.91%,RSD为0.34%（n=9）。结论：该方法可快速筛选出合适的含量测定指标;所建立的测定方法简便、准确,可作为神香草水提物中迷迭香酸的含量测定方法。

OBJECTIVE： To study the main components of Hyssopus officinalis water extract, and determine the content. METHODS： LC-MS method was used to analysis the main components in 1-1. officinalis water extract. LC-MS condition was as fol- lows： the chromatographic column was waters 5 C18-DAQ column with mobile phase consisted of acetonitrile-0.2% acetic acid （gra- dient elution） at the flow rate of 1.0 ml/min; the column temperature was 30℃ and detection wavelength was set at 360 nm; the sample size was 10 μl; the sample was analyzed by ESI in negative-ion mode; ion source temperature was 400 ℃ and m/z rang was from 100-1 000 Da. The rosmarinic acid was selected as indicator component and RP-HPLC was conducted to determine the content. RP-HPLC condition was as follows： the chromatographic column was Phenomenex C18 column with mobile phase consisted of acetonitrile-0.2% acetic acid （25 ： 75, V/V） at the flow rate of 1.0 ml/min; the detection wavelength was 330 nm, and column temperature was 30 ℃ ; the sample size was 10 μl. RESULTS： LC-MS analysis showed that rosmarinic acid was the main compo- nent of H. officinalis water extract. The linear range of rosmarinic acid were 4.52-22.6 0μg/ml（r=0.999 9） with an average recov- ery of 99,91% （RSD=0.34% , n=9） ; RSDs of precision, stability and reproducibility tests were all lower than 2%. CONCLU- SIONS： The method can analyze H. officinalis water extract rapidly, and screen suitable content determination indicator. Estab- lished RP-HPLC method is simple, rapid, accurate and reproducible. It can be used for the content determination of H. officinalis water extract.