柱前衍生HPLC/FLD-APCI/MS法同时测定藏药悬钩木中6种三萜酸类成分的含量  

Content Determination of 6 Kinds of Triterpene Acid in Tibetan Medicine Rubus biflorus by Pre-column Derivatization HPLC/FLD-APCI/MS

作　　者：马志良 赛桑杰 多杰 MA Zhiliang;SAI Sangjie;DUO Jie(State Key Laboratory of Tibetan Medicine Research and Development/Qinghai Province Key Laboratory of Tibetan Medicine Research and Development, Xining810016,China;Qinghai Tibetan Medicine Research Institute,Xining 810016,China)

机构地区：[1]藏药新药开发国家重点实验室/青海省藏药新药开发重点实验室,西宁810016 [2]青海省藏医药研究院,西宁810016

出　　处：《中国药房》2019年第16期2243-2247,共5页China Pharmacy

基　　金：藏药新药开发国家重点实验室项目(No.2015DQ87-0717);青海省科技计划项目(No.2017-ZJ-Y15)

摘　　要：目的:建立同时测定藏药悬钩木中山楂酸、科罗索酸、桦木酸、路路通酸、齐墩果酸、熊果酸等6种三萜酸类成分含量的方法。方法:采用柱前衍生高效液相色谱-荧光检测/质谱联用法。选择2-(7H-二苯并[a,g]咔唑)乙基对甲苯磺酸酯为荧光衍生化试剂。色谱柱为Hypersil C18,流动相为5%乙腈水溶液-乙腈(梯度洗脱),流速为1.0 mL/min,荧光激发波长为300 nm,发射波长为395 nm,柱温为35℃,进样量为10μL;采用大气压化学电离源、正离子模式,喷雾压力为60 psi,干燥气流量为9 L/min,干燥气温度为350℃,气化温度为450℃,毛细管电压为3 500 V。结果:山楂酸、科罗索酸、桦木酸、路路通酸、齐墩果酸、熊果酸检测质量浓度线性范围均为0.025~6.4μg/mL(r≥0.999 6);定量限分别为5.11、4.78、4.42、4.22、4.29、4.51 ng/mL,检测限分别为1.42、1.27、1.30、1.28、1.16、1.22 ng/mL;精密度试验的RSD均小于5%,稳定性、重复性试验的RSD均小于2%(路路通酸未检出);加样回收率分别为97.90%~100.55%(RSD=1.00%,n=6)、97.95%~102.95%(RSD=1.74%,n=6)、96.00%~101.20%(RSD=2.00%,n=6)、93.25%~104.20%(RSD=4.25%,n=6)、92.20%~103.30%(RSD=3.58%,n=6)、97.80%~103.50%(RSD=2.03%,n=6)。结论:该方法准确、可靠,专属性好,可用于同时测定藏药悬钩木中6种三萜酸类成分的含量。OBJECTIVE:To establish the method for content determination of 6 kinds of triterpene acids such as haw acid,corosolic acid,betula acid,betulonic acid,oleanolic acid and ursolic acid in Tibetan medicine Rubus biflorus. METHODS:Pre-column devrivatization HPLC-FLD-APCI/MS method was adopted. 2-(7 H-dibenzo[a,g]carbazol-7-yl) ethyl-4-methylbenzenesulfonate was used as the pre-column derivatization reagent.Hypersil C18 column was used with the mobile phase consisted of 5%acetonitrile water solution-acetonitrile(gradient elution)at the flow rate of 1.0 mL/min. The excitation wavelength of fluorescence was 300 nm and the emission wavelength was 395 nm. The column temperature was 35 ℃,and sample size was 10 μL. Under atmospheric-pressure chemical ionization(APCI)source in positive-ion mode,pressure was 60 psi,the drying gas flow rate was 9 L/min,the dry gas temperature was 350 ℃,the gasification temperature was 450 ℃,and the capillary voltage was 3 500 V.RESULTS:The linear range of haw acid,corosolic acid,betula acid,betulonic acid,oleanolic acid and ursolic acid were 0.025-6.4μg/mL(r≥0.999 6). The quantitative limits were 5.11,4.78,4.42,4.22,4.29,4.51 ng/mL;and detection limits were 1.42,1.27,1.30,1.28,1.16,1.22 ng/mL,respectively. RSD of precision test was less than 5%,stability and repeatability tests were all less than 2%(no betulonic acid detected). The recovery rates were 97.90%-100.55%(RSD=1.00%,n=6),97.95%-102.95%(RSD=1.74%,n=6),96.00%-101.20%(RSD=2.00%,n=6),93.25%-104.20%(RSD=4.25%,n=6),92.20%-103.30%(RSD=3.58%,n=6),97.80%-103.50%(RSD=2.03%,n=6),respectively. CONCLUSIONS:The method is accurate,reliable and exclusive,and can be used for simultaneous determination of 6 kinds of triterpene acids in Tibetan medicine R. biflorus.

关 键 词：悬钩木 三萜酸 山楂酸 科罗索酸 桦木酸 路路通酸 齐墩果酸 熊果酸 柱前衍生化 高效液相色谱-荧光检测/质谱联用 含量测定 

分 类 号：R284.1[医药卫生—中药学] R917[医药卫生—中医学]