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建立辽细辛中2个主要的直链酰胺成分十二碳四烯酰胺A与十二碳四烯酰胺B的定性与定量分析方法,并测定其在42份辽细辛(37份不同年份收集的北细辛及5份汉城细辛)中的含量。利用HPLC-IT-TOF-MS/MS技术结合对照品鉴定北细辛甲醇提取液中的十二碳四烯酰胺A与B;对2种成分的含量测定采用超高效液相色谱-二极管阵列检测器(UPLC-PDA),ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7μm),流动相为纯水-乙腈,梯度洗脱,柱温45℃,检测波长254 nm。结果表明,HPLC-IT-TOF-MS/MS鉴别细辛中十二碳四烯酰胺A、B的准分子离子[M+H]+为m/z 248.20;2种成分在UPLC-PDA上分离度良好,在检测范围内呈良好线性,平均加样回收率为97.90%和99.86%。在所测定辽细辛样品中,十二碳四烯酰胺A的含量为0.11~3.89 mg·g-1,十二碳四烯酰胺B为0.24~6.65 mg·g-1。含量测定结果显示随着贮藏时间的延长,十二碳四烯酰胺A与十二碳四烯酰胺B的含量均呈现降低的趋势,与2013年收集样品相比,2002~2003年收集样品中两者的平均含量分别降低34%和36%;这2种成分在汉城细辛中的平均含量[十二碳四烯酰胺A:(0.78±0.52)mg·g-1;十二碳四烯酰胺B:(1.69±0.83)mg·g-1]均显著低于北细辛[十二碳四烯酰胺A:(1.59±0.75)mg·g-1;十二碳四烯酰胺B:(2.90±1.17)mg·g-1](P<0.05);辽细辛地上部分十二碳四烯酰胺A的含量为0.11~0.33 mg·g-1,十二碳四烯酰胺B含量为0.24~0.60 mg·g-1,两者的含量均明显低于同一植株的地下部分(分别为0.73~3.89,2.11~6.24 mg·g-1)。本方法快速简便、结果准确,对2种直链酰胺类成分可达到良好的分离并满足含量测定的要求,可用于辽细辛药材中这2种成分的定性与定量分析,为辽细辛药材质量控制方法的进一步提高提供依据。
To establish a qualitative and quantitative method for the determination of two main linear chain amides, Dodecatetraenoamide A and Dodecentetraenoamide B, in Asarum heterophylla, and determine their bioavailability in 42 Aspartame (collected in 37 different years Of Asarum and 5 parts of Asarum in Seoul). Dodecatetraenoamide A and B were identified by HPLC-IT-TOF-MS / MS combined with reference substance. The contents of two components were determined by ultra performance liquid chromatography-diode array (UPLC-PDA) and ACQUITY UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 μm). The mobile phase consisted of pure water-acetonitrile. The mobile phase was eluted with a gradient of 45 ℃. The detection wavelength was 254 nm. The results showed that [M + H] + was identified as m / z 248.20 by HPLC-IT-TOF-MS / MS. The two components were separated on UPLC-PDA Degree of good, within the detection range showed a good linearity, the average recovery was 97.90% and 99.86%. In the tested asarum samples, the content of dodecentetraenoamide A was 0.11-3.89 mg · g-1, and the amount of dodeca-tetraenamide B was 0.24-6.65 mg · g-1. The content of decadientetraamide A and dodecentetraenamide B showed a trend of decreasing with the prolongation of storage time. Compared with the samples collected in 2013, the contents of two of the samples collected in 2002-2003 The average content of the two components decreased by 34% and 36%, respectively. The average contents of these two components in Asarum were significantly higher than that of the control group [Dodecatetraenol A: (0.78 ± 0.52) mg · g-1) B: (1.69 ± 0.83) mg · g-1] were significantly lower than that of asarum [Dodecatetraenol A: (1.59 ± 0.75) mg · g-1; Dodecatetraenoamide B: ± 1.17) mg · g-1] (P <0.05). The content of Codontetraamide A in the above-ground part of Liaozishan was 0.11-0.33 mg · g-1, the content of Codontetraamide was 0.24 ~ 0.60 mg · g-1, both of which were significantly lower than those of the same plant (0.73-3.89 and 2.11-6.24 mg · g-1, respectively). The method is rapid and convenient with accurate results and can achieve good separation and content determination of two linear amides and can be used for qualitative and quantitative analysis of the two components in the Liao Xushen herbs. Quality control methods to further improve provide the basis.