摘要
目的基于中药质量标志物(qualitymarker,Q-Marker)概念,采用超高效液相色谱-串联质谱(UPLC-MS/MS)方法建立同时测定血浆中热毒宁注射液13个潜在Q-Marker含量的方法,并用于大鼠体内血药浓度测定及药动学研究,以证明其体外-体内传递过程的可测性。方法采用Waters AQUITY UPLC C18色谱柱(100 mm×2.1 mm,1.7μm),流动相为0.1%甲酸水-乙腈梯度洗脱,体积流量0.3 mL/min;柱温为35℃,自动进样器温度为4℃;进样量为2μL。以氯霉素为内标,采用电喷雾离子源(ESI),负离子源,多反应离子检测(multiple reaction monitoring,MRM)模式,同时定量血浆中13个Q-Marker(栀子苷、京尼平苷酸、京尼平龙胆双糖苷、山栀苷、绿原酸、新绿原酸、隐绿原酸、异绿原酸A、异绿原酸B、4,5-二-O-咖啡酰奎宁酸、裂环氧化马钱子苷、裂环马钱子酸、獐牙菜苷)含量。采用Phoenix WinNonlin8.1软件非房室模型计算药动学参数。结果13个化合物在检测范围内线性关系良好。大鼠血浆中内源性物质不干扰测定,该方法回收率、基质效应、精密度、准确度和稳定性均符合生物样品的测定要求。尾iv热毒宁注射液后在大鼠血浆中可检测到上述13个化合物,裂环马钱子酸在血浆中暴露量最大,栀子苷和绿原酸次之。各化合物在体内半衰期较短,消除速率快;表观分布容积提示所有化合物呈广泛组织分布。结论该方法可以同时定量血浆中热毒宁注射液的13个成分,证明了上述成分体外-体内传递过程的可测性;药动学研究表明上述成分还具有血浆-组织/靶器官传递性,可作为热毒宁注射液安全性和有效性评价的潜在Q-Marker。
Objective Based on the concept of traditional Chinese medicine quality marker(Q-Marker),a method for simultaneous determination of 13 potential Q-Markers in plasma was established by ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).The method was also used to determine the plasma concentration of 13 potential Q-Markers and their pharmacokinetics in health rats given Reduning Injection(热毒宁注射液)via tail vein in order to prove the detectability of transfer process in vitro and in vivo.Methods Waters AQUITY UPLC C18 column(100 mm×2.1 mm,1.7μm)was used.The mobile phase was 0.1%formic acid water-acetonitrile gradient elution,the flow rate was 0.3 m L/min,the column temperature was 35℃,the temperature was 4℃,and the injection volume was 2μL.Using chloramphenicol as an internal standard,13 Q-Markers in plasma(geniposide,geniposidic acid,genipin-1-β-D-gentiobioside,shanzhiside,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,isochlorogenic acid B,4,5-dicaffeoylquinic acid,secoxyloganin,secologanic acid,and sweroside)were quantified simultaneously by using electrospray ion source(ESI),negative ion source,and multiple reaction monitoring(MRM).The non-compartment model was used to calculate the pharmacokinetic parameters in Phoenix Win Nonlin8.1 software.Results The 13 compounds showed good linear relationship in the detection range.The recovery rate,matrix effect,precision,accuracy and stability of the method all match the requirements of biological samples detection.The results showed that the above 13 compounds could be detected in the plasma of rats injected with Reduning Injection via tail vein.The exposure concentration of secologanic acid was the highest,followed by concentration of geniposide and chlorogenic acid.All compounds owned short half-life and quick elimination rate in vivo,and large apparent distribution volume which indicated that they were widely distributed in tissues.Conclusion The method can simultaneously quantify 13 compounds of Reduning Injection in plasma,which proves the detectability of the above components’transfer process from in vitro to in vivo.The pharmacokinetic study showed that the 13 compounds also had plasma tissue/target organ transfer property,which can be used as potential Q-Marker for safety and efficacy evaluation of Reduning Injection.
作者
王璨
王保和
黄宇虹
李自强
张涵
WANG Can;WANG Bao-he;HUANG Yu-hong;LI Zi-qiang;ZHANG Han(Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300250,China)
出处
《中草药》
CAS
CSCD
北大核心
2021年第9期2653-2664,共12页
Chinese Traditional and Herbal Drugs
基金
国家“重大新药创制”科技重大专项资助项目(2018ZX09734-002)
天津市自然科学基金项目(18JCQNJC83800)
天津市科技计划项目(17ZXXYSY00060)。
