Guo Na, Ding Weimin, Wang Yu, Hu Zhiwei, Wang Ziming, Wang Yang
School of Municipal and Environmental Engineering, Harbin Institute of Technology , Harbin , China .
Pharm Biol. 2014 May;52(5):637-45. doi: 10.3109/13880209.2013.863946. Epub 2014 Jan 30.
Salidroside and its metabolite p-tyrosol are two major phenols in the genus Rhodiola L. (Crassulaceae). They have been confirmed to possess various pharmacological properties and are used for the prophylaxis and therapeutics of many diseases. Several analytical methods have been developed for the determination of the two compounds in plant materials and biological plasma matrices. However, these methods are not optimal for biological samples containing complex organic interferences, such as liver and brain tissues.
This study aimed to further develop and validate a simple and specific LC-MS/MS method for the determination of salidroside and its metabolite p-tyrosol in rat liver tissues using paracetamol as the internal standard (IS).
Salidroside and p-tyrosol with the IS paracetamol and liver tissues were used as model compounds and biological samples. Samples were processed by protein precipitation (PP) with methanol, the supernatant was dried under nitrogen and the residue was reconstituted in a mobile phase that consisted of a mixture of acetonitrile and water (1:9, v/v). Salidroside and p-tyrosol were detected in negative mode under multiple reaction monitoring (MRM) by a triple quadrupole tandem mass spectrometer coupled with electrospray ionization.
Standard curves were linear over the concentration range of 50-2000 ng/mL with correlation coefficients of 0.995 or better for both salidroside and p-tyrosol. The intra- and inter-day accuracy for salidroside ranged between 104.90 and 112.73% with a precision of 3.51-14.27%. For p-tyrosol, the intra- and inter-day accuracy was between 92.38 and 100.59%, and the precision was 8.54% or less. The stability data showed that no significant degradation occurred under the experimental conditions. The recoveries were 111.44, 108.10, and 102.00% for salidroside at concentrations of 50, 500 and 2000 ng/mL, respectively, and were 105.44, 105.50, and 113.04% for tyrosol at concentrations of 50, 500 and 2000 ng/mL, respectively. The matrix effects were 83.85-92.45% for salidroside and 85.61-92.49% for p-tyrosol at three QC levels. This method was successfully applied to a liver tissue distribution study of salidroside and its metabolite p-tyrosol in rats.
This newly established method is validated as simple, reliable and accurate. It can be used as a valid analytical method for the intrinsic quality control of biological matrices, especially tissue samples.
红景天苷及其代谢产物对羟基苯乙醇是景天科红景天属植物中的两种主要酚类物质。它们已被证实具有多种药理特性，可用于多种疾病的预防和治疗。已开发出几种分析方法用于测定植物材料和生物血浆基质中的这两种化合物。然而，这些方法对于含有复杂有机干扰物的生物样品（如肝脏和脑组织）并非最佳选择。
本研究旨在进一步开发并验证一种简单、特异的液相色谱-串联质谱法，以对乙酰氨基酚为内标物（IS），用于测定大鼠肝脏组织中的红景天苷及其代谢产物对羟基苯乙醇。
将红景天苷、对羟基苯乙醇与内标物对乙酰氨基酚以及肝脏组织用作模型化合物和生物样品。样品采用甲醇进行蛋白沉淀（PP）处理，上清液在氮气下吹干，残渣用由乙腈和水（1:9，v/v）组成的流动相复溶。采用配备电喷雾电离的三重四极杆串联质谱仪，在多反应监测（MRM）的负离子模式下检测红景天苷和对羟基苯乙醇。
标准曲线在50 - 2000 ng/mL浓度范围内呈线性，红景天苷和对羟基苯乙醇的相关系数均为0.995或更佳。红景天苷的日内和日间准确度在104.90%至112.73%之间，精密度为3.51% - 14.27%。对于对羟基苯乙醇，日内和日间准确度在92.38%至100.59%之间，精密度为8.54%或更低。稳定性数据表明，在实验条件下未发生显著降解。红景天苷在50、500和2000 ng/mL浓度下的回收率分别为111.44%、108.10%和102.00%，对羟基苯乙醇在50、500和2000 ng/mL浓度下的回收率分别为105.44%、105.50%和113.04%。在三个质量控制水平下，红景天苷的基质效应为83.85% - 92.45%，对羟基苯乙醇的基质效应为85.61% - 92.49%。该方法成功应用于大鼠红景天苷及其代谢产物对羟基苯乙醇的肝脏组织分布研究。
新建立的方法经验证简单、可靠且准确。它可作为生物基质尤其是组织样品内在质量控制的有效分析方法。
