女贞子饮片质量评价研究

目的研究建立女贞子饮片质量评价标准。方法以红景天苷和酪醇为对照品进行TLC鉴别,同时进行HPLC含量测定,采用Luna C18柱,流动相:乙腈-0.05%磷酸(3∶97),流速:1.0mL·min-1,检测波长:224nm。结果女贞子酒蒸后TLC图谱有显著差别,红景天苷和酪醇的含量显著增加。结论TLC方法简便、专属性强,HPLC方法测定精密度高、重复性好,为女贞子饮片的质量评价方法提供了试验依据。     

红景天提取物的质量标准研究

目的对红景天提取物进行质量标准研究。方法采用薄层色谱法对红景天苷和酪醇进行定性鉴别；采用高效液相色谱法测定红景天提取物中红景天苷的含量。结果在TLC色谱中能检测出红景天苷和酪醇；HPLC定量方法的线性范围为16．14～161．4μg，平均回收率为100．02％。结论本研究建立的定性、定量方法准确可行，重复性好，可作为红景天提取物的质量标准。     

SPE-HPLC测定注射用红景天(冻干)中红景天苷和酪醇的含量

目的:测定注射用红景天(冻干)中红景天苷和酪醇的含量。方法:选用C₁₈固相萃取小柱对样品进行纯化后用高效液相色谱法测定红景天苷和酪醇含量。Irregular-H C₁₈柱(4.6 mm×250 mm,10μm),流动相甲醇-乙腈-0.06%磷酸(10∶10∶80),流速1.0 mL.min^-1,检测波长275 nm,柱温30℃。结果:红景天苷在2.24~22.4μg、酪醇在0.856~8.56μg呈现良好的线性相关性,r分别为0.999 7,0.999 6,平均回收率分别为101.3%,99.8%。结论:本方法简便快捷,分离度好,结果准确可靠。     

An LC–MS/MS method for the determination of salidroside and its metabolite p-tyrosol in rat liver tissues

Context: 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.

Objective: 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).

Materials and methods: 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.

Results: 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.

Discussion and conclusion: 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.     

LC-MS/MS法测定比格犬血浆中红景天苷和酪醇的浓度及其药动学研究

目的:建立LC-MS/MS法测定比格犬血浆中红景天苷及其代谢物酪醇的浓度,并应用于药动学研究。方法:血浆样品经氯仿、苯酚和异戊醇混合溶剂萃取,Shimadzu Shim-pack Velox PFPP色谱柱(100 mm×2.1 mm,1.8μm)分离,以水(A相)-含20%乙腈的甲醇溶液(B相)为流动相,梯度洗脱(0~1 min,12%B;1~2.5 min,12%~85%B;2.5~3.5 min,85%B;3.5~3.6 min,85%~12%B;3.6~5 min,12%B)5 min,流速为0.3 ml/min,柱温40℃。采用电喷雾离子源(ESI),负离子多反应监测模式(MRM)进行质谱监测,离子对:红景天苷m/z 299.1→118.9、酪醇m/z 137.0→105.9、内标天麻素m/z 285.1→122.9。测定比格犬口服和静脉注射红景天苷后的主要药动学参数。结果:红景天苷在10~10000 ng/ml,酪醇在1~1000 ng/ml范围内线性良好,日内和日间精密度RSD均<12.9%。口服给药后,红景天苷和酪醇的AUC0~t分别为(19600±4870)和(946±188)ng·h·ml^-1,AUC_0~∞分别为(19700±4880)和(2280±1780)ng·h·ml^-1,c_max分别为(8890±2650)和(132±44.3)ng/ml,t_1/2分别为(1.85±0.69)和(40.3±45.1)h;静脉注射给药后,红景天苷和酪醇的AUC0~t分别为(5960±882)和(127.0±32.4)ng·h·ml^-1,AUC_0~∞分别为(6000±887)和(157.0±31.6)ng·h·ml^-1,c_max分别为(9210±1900)和(42.0±12.3)ng/ml,t_1/2分别为(0.976±0.156)和(4.46±2.42)h。结论:本方法简单、高效,专属性好,灵敏度高,可用于红景天苷及其代谢物酪醇在比格犬体内的药动学研究。     

Tyrosol as a Neuroprotector: Strong Effects of a “Weak” Antioxidant

The use of neuroprotective agents for stroke is pathogenetically justified, but the translation of the results of preclinical studies of neuroprotectors into clinical practice has been a noticeable failure. One of the leading reasons for these failures is the one-target mechanism of their activity. p-Tyrosol (Tyr), a biophenol, is present in a variety of natural sources, mainly in foods, such as olive oil and wine. Tyr has a wide spectrum of biological activity: antioxidant, stress-protective, anti-inflammatory, anticancer, cardioprotective, neuroprotective and many others. This review analyzes data on the neuroprotective, antioxidant, anti-inflammatory, anti-apoptotic and other kinds of Tyr activity as well as data on the pharmacokinetics of the substance. The data presented in the review substantiate the acceptability of tyr as the basis for the development of a new neuroprotective drug with multitarget activity for the treatment of ischemic stroke. Tyr is a promising molecule for the development of an effective neuroprotective agent for use in ischemic stroke.