基于UPLC-MS技术分析马钱子-甘草药对配伍汤液不同相态中毒效物质的变化规律

Changes of toxic substance in different phase of compatibility decoction Strychni Semen and Glycyrrhizae Radix et Rhizoma based on UPLC-MS technique

目的以相态角度为出发点,研究马钱子-甘草配伍减毒的作用规律。方法采用自然沉降法、高速离心法及层级透析技术对马钱子单煎液、马钱子-甘草配伍共煎液进行相态拆分研究;建立毒效物质UPLC-MS测定法,对比研究4个不同相态中马钱子碱和士的宁量分布的差异,分析毒效物质量的变化规律。结果配伍汤液可拆分为真溶液组、胶体溶液组、混悬液组及沉积物组4个不同相态;在选定的色谱条件下马钱子碱和士的宁均得到良好分离,分别在0.020~2.008、0.022~2.236μg线性关系良好(r均为0.999 8),回收率为96.84%、97.62%,表明方法学良好;测定单煎液不同相态中马钱子碱和士的宁量的分布呈现相同变化规律,即胶体组>溶液组>沉积物组>混悬组,配伍汤液不同相态马钱子碱和士的宁也同样呈现相同的分布规律,即沉积物组>胶体组>混悬组>溶液组。结论马钱子经甘草配伍后其毒效物质的量均有降低,毒性最强的士的宁的量降低尤为明显,且主要集中在沉积物组分;不同相态中毒效物质变化规律的研究表明,配伍甘草后的沉积物组分的缓释作用,是实现减毒增效关键因素。

Objective To study the mechanism of attenuated Strychni Semen（SS） and Glycyrrhizae Radix et Rhizoma（GRR） compatibility with the phase angle as the starting point.Methods The phase separation of SS single decoction and compatibility decoction was carried out by natural sedimentation,dialysis and high-speed centrifugation.UPLC-MS assay was used to study the differences in the distribution of brucine and strychnine in four different phases,and the change of the amount of toxic substances was analyzed.Results The compatibility of the decoction can be divided into four groups： true solution group,colloidal solution group,suspension group,and sediment group.Brucine and strychnine were well separated under the selected chromatographic conditions.The linear relationship was good in the range of 0.020—2.008 and 0.022—2.236 μg（r = 0.999 8）,and the recovery was 96.84% and 97.62%,which indicated that the method was good.The distribution of brucine and strychnine in the different phase of the single decoction showed the same variation regularity,colloid group solution group sediment group suspended group; Compatibility decoction of different phase of brucine and strychnine also showed the same distribution,sediment group colloidal group suspension group solution group.Conclusion The contents of toxic substances in SS were decreased after compatibility with GRR,especially for the strongest toxicity of strychnine,which mainly concentrated in the sediment components.Studies on the changes of toxic substances in phase states indicated that the sustained release effect of the sediments components after compatibility with GRR is the key factor of reducing toxicity and improving efficiency.