7-乙基-10-羟基喜树碱脂质体在大鼠体内的代谢和排泄

目的考察7-乙基-10-羟基喜树碱(SN-38)脂质体经静脉注射后,在大鼠尿液、粪便中的代谢产物以及以SN-38原形药物排泄的量。方法大鼠尾静脉单次给予2.77 mg/kg SN-38脂质体,分别于0～6、6～12、12～24、24～48 h分段收集尿液、粪便,采用UPLC/Q-TOFMS法对SN-38脂质体在大鼠尿液、粪便中的代谢产物进行鉴定,并且建立HPLC法,用于大鼠尿液及粪便样品中SN-38原形药物的排泄量的测定。结果 SN-38脂质体的在大鼠体内的代谢产物经鉴定为SN-38G。48 h内脂质体组共有1.57%的原形药物经过尿液排出,共有12.94%的SN-38原形药物经过粪便排出。结论 SN-38脂质体只有少部分以原形药物经尿液和粪便排出体外。     

前胡香豆素类提取物的UPLC/Q-TOF-MS分析及其初步药效学研究

目的建立UPLC/Q-TOF-MS法分析前胡香豆素类提取物的化学成分,并对其初步药效学进行研究。方法采用Acquity UPLC BEH C_(18)色谱柱;以0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱;检测波长321 nm;采用电喷雾正离子化模式(ESI~+),扫描范围m/z 100~1 000;利用链脲佐菌素(STZ)诱导的2型糖尿病大鼠模型,对前胡香豆素类提取物的调脂降糖作用进行初步药效学研究。结果前胡香豆素类提取物中的qianhucoumarinD(1)、peucedanocoumarinⅡ(2)、白花前胡甲素(3)、peucedanocoumarin I(4)、白花前胡乙素(5)、praeuptorin E(6)得到良好的分离与鉴定。初步药效学实验结果表明,前胡香豆素类提取物能显著降低2型糖尿病大鼠空腹血糖、胆固醇、三酰甘油、低密度脂蛋白及肝脏指数(P0.05),升高高密度脂蛋白(P0.05),提示前胡香豆素类提取物能改善2型糖尿病大鼠糖与脂质代谢,发挥治疗2型糖尿病的作用。结论建立了前胡香豆素类的UPLC/Q-TOF-MS的分离鉴定方法;初步药效学研究表明前胡香豆素类提取物具有显著的调脂及降糖作用。     

高良姜黄酮提取物特征谱分析及成分确认

目的:建立不同批次高良姜提取物的特征谱并进行成分鉴定。方法:采用HPLC分析高良姜提取物的特征指纹谱。SunfireTMC18色谱柱,流动相乙腈-0.1%的甲酸溶液梯度洗脱,分析时间60 min,检测波长208 nm,柱温30℃,流速1.0 mL.min-1。采用超高效液相色谱串联四级杆飞行时间质谱联用技术(UPLC/Q-TOF MS)进行分析,对主要成分进行鉴定。结果:特征谱共标出10个共有峰。鉴定出提取物中12个化合物。结论:采用HPLC特征指纹谱能有效控制高良姜提取物的质量。     

UHPLC-Q-TOF／MS技术应用于中药旱莲草化学成分研究

目的通过超高效液相色谱-四极杆飞行时间串联质谱（UHPLC-Q-TOF/MS）对中药旱莲草的化学成分进行系统、全面研究。方法采用Agilent SB-C18（2.1 mm×100 mm,1.8μm）柱,流动相为乙腈-甲酸溶液系统梯度洗脱,质谱采用ESI正离子模式,分别对桂林阳朔、桂林荔蒲、湖南邵阳、南宁和湖北襄樊五个产地的旱莲草药材进行分析。结果该方法共鉴别出旱莲草中29个化学成分。结论本实验为建立旱莲草药材的质量控制标准提供了参考依据。     

2种溶血磷脂的飞行时间质谱裂解特征研究

目的研究sn-甘油-1-花生苷-3-磷脂酰胆碱和sn-甘油-1-花生苷-2-磷脂酰胆碱的质谱裂解特征,对其进行区分,探讨该类化合物的质谱裂解规律。方法建立Q-TOF/MS法对sn-甘油-1-花生苷-3-磷脂酰胆碱和sn-甘油-1-花生苷-2-磷脂酰胆碱进行质谱检测,观察其在不同碰撞能量（5 eV、15 eV、25 eV、35 eV、45 eV）下产生的碎片离子。结果在不同的碰撞能量下,sn-甘油-1-花生苷-3-磷脂酰胆碱产生碎片离子m/z104,125,147,184,317,361和485,而sn-甘油-1-花生苷-2-磷脂酰胆碱产生碎片离子m/z104,125,147,184,361和485。由此推测出溶血磷脂的裂解规律,并对sn-甘油-1-花生苷-3-磷脂酰胆碱和sn-甘油-1-花生苷-2-磷脂酰胆碱进行了区分。结论 Q-TOF/MS法可以有效地对同分异构体化合物sn-甘油-1-花生苷-3-磷脂酰胆碱和sn-甘油-1-花生苷-2-磷脂酰胆碱进行结构解析。     

Characterization of the constituents in rat biological fluids after oral administration of Fufang Danshen tablets by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry

An ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry method was established to detect as many constituents in rat biological fluids as possible after oral administration of Fufang Danshen tablets (FDTs). A C18 column (1.8 μm particle size) was adopted to separate the samples, and mass spectra were acquired in both negative and positive modes. First, the fingerprints of FDTs were established, resulting in 43 components being detected within 25 min. Among these compounds, 37 were tentatively identified by comparing the retention times and mass spectral data with those of reference standards and the reference literature; the other 6 components were tentatively assigned solely based on the MS data. In vivo, 14 components and 8 metabolites of FDT were observed in plasma, and 12 components of FDT were detected in urine. Tanshinaldehyde, danshexinkun B, a glycine conjugate of danshensu and a methylated conjugate of danshexinkun B were newly detected in rat biological fluids. This study developed a high-speed and sensitive method that was successfully utilized for screening the active ingredients of a Chinese medical formula and provided helpful chemical information for further pharmacology and active mechanism research on Chinese medicine.     

桂枝茯苓胶囊中三萜酸类成分的UPLC/Q-TOF-MS指纹图谱研究

目的建立桂枝茯苓胶囊(GFC)中三萜酸类成分UPLC/Q-TOF-MS指纹图谱方法,为评价GFC的质量提供新方法。方法采用UPLC分离三萜酸类成分,用Q-TOF-MS检测,建立UPLC/Q-TOF-MS指纹图谱。结果得到灵敏度、选择性和专属性良好的GFC中三萜酸类成分UPLC/Q-TOF-MS指纹图谱,确定了26个共有峰,其中3、5～18、20、23、24共18个峰来自于茯苓,2号峰来自于白芍和牡丹皮,4号峰来自于茯苓、牡丹皮、白芍、桂枝,19号峰来自于牡丹皮、白芍和桂枝,21号峰来自于牡丹皮和白芍,22、25号峰来自于茯苓、牡丹皮、桃仁、白芍、桂枝,26号峰来自于桂枝、白芍、桃仁。10批GFC指纹图谱相似度在0.90以上。UPLC-Q-TOF-MS共鉴定出16个成分,分别为16α-羟基松苓新酸、16α-羟基-栓菌酸、3-酮基-6,16α-二羟基-羊毛甾-7,9(11),24-三烯-21酸、去氢土莫酸、土莫酸、3-酮基-6,16α-二羟基-羊毛甾-8,24-二烯-21酸、依布里酸、猪苓酸C、3-表去氢土莫酸、3-O-乙酰基-16α-羟基松苓新酸、3-表去氢茯苓酸、3-O-乙酰基-16α-羟基-栓菌酸、去氢茯苓酸、茯苓酸、松苓新酸、去氢齿孔酸。结论该方法准确、快速,具有较好的精密度、重复性和稳定性,适用于GFC的质量控制。     

Investigation of 6-O-methyl-scutellarein metabolites in rats by ultra-flow liquid chromatography/quadrupole-time-of-flight mass spectrometry

Context Scutellarin (1) has been widely used in China to treat acute cerebral infarction and paralysis induced by cerebrovascular diseases. However, scutellarin (1) has unstable metabolic characteristics.

Objective The metabolic profile of 6-O-scutellarein was studied to determine its metabolic stability in vivo.

Materials and methods In this study, a method of UFLC/Q-TOF MS was used to study the 6-O-methyl-scutellarein metabolites in rat plasma, urine, bile and faeces after oral administration of 6-O-methyl-scutellarein (3). One hour after oral administration of 6-O-methyl-scutellarein (3) (34?mg/kg), approximately 1?mL blood samples were collected in EP tubes from all groups. Bile, urine and faeces samples were collected from eight SD rats during 0–24?h after oral administration. The mass defect filtering, dynamic background subtraction and information dependent acquisition techniques were also used to identify the 6-O-methyl-scutellarein metabolites.

Results The parent compound 6-O-methyl-scutellarein (3) was found in rat urine, plasma, bile and faeces. The glucuronide conjugate of 6-O-methyl-scutellarein (M1, M2), diglucuronide conjugate of 6-O-methyl-scutellarein (M3), sulphate conjugate of 6-O-methyl-scutellarein (M4), glucuronide and sulphate conjugate of 6-O-methyl-scutellarein (M5), methylated conjugate of 6-O-methyl-scutellarein (M6) were detected in rat urine. M1, M2 and M3 were detected in rat bile. M1 was found in rat plasma and M7 was detected in faeces.

Discussion and conclusion Because the parent compound 6-O-methyl-scutellarein (3) was found in rat urine, plasma, bile and faeces, we speculate that 6-O-methyl-scutellarein (3) had good metabolic stability in vivo. This warrants further study to develop it as a promising candidate for the treatment of ischemic cerebrovascular disease.     

基于UPLC/QTOF-MS的代谢组学研究辛伐他汀对不同性别小鼠尿液中代谢物的影响

目的:基于代谢组学技术研究辛伐他汀对小鼠尿液中脂类代谢物的影响。方法:收集给予药物辛伐他汀后不同天数的小鼠尿液样品,采用超高效液相色谱-高分辨飞行时间质谱(UPLC/Q-TOF MS)联用,Acquity BEH C₁₈色谱柱,流动相为水-乙腈,梯度洗脱,电喷雾离子源,负离子检测模式,通过数据采集进行代谢轮廓分析。采用MarkerLynxXS软件读取总离子流色谱图中峰信息,利用主成分分析和正交偏最小二乘法-判别分析对样品进行分类。筛选出候选的差异代谢物后,通过一级、二级质谱碎片信息,比对数据库鉴别差异代谢物的结构。结果:给药辛伐他汀后,雌鼠和雄鼠的体质量增长速率存在差异,尿液中的代谢物变化有所不同。鉴定出棕榈酸、胆甾烷五醇、羟睾酮葡糖醛酸苷、硝基亚油酸胆固醇、磷脂酸和磷脂酰乙醇胺共6个脂类代谢差异物,呈现出对体内脂类代谢的影响,在雌雄个体中呈现不同的变化规律。结论:研究结果提示辛伐他汀的临床用药需要考虑性别差异的因素。     

Simultaneous analysis of 23 illegal adulterated aphrodisiac chemical ingredients in health foods and Chinese traditional patent medicines by ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry

This paper presents an application of ultra high-performance liquid-chromatography-quadrupole-TOF high resolution mass spectrometry (UHPLC-Q-TOF HRMS) for simultaneous analysis of 23 illegal adulterated aphrodisiac type chemical ingredients in health foods and Chinese Traditional Patent Medicines (CTPMs). The mass spectrometer was operated in Information Dependent Acquisition (IDA) mode, which provides crucial information for the elemental composition analysis, structure elucidation and quantitative analysis simultaneously. Quantitative analysis was performed using the peak areas of the precursor ions in the XICs. The method validation included assessment of selectivity, sensitivity, calibration curve, accuracy, precision, recovery, matrix effect and stability. The results show good linear relationship with the concentrations of the analytes over wide concentration ranges (e.g., 0.05–10 μg/g for sildenafil) as all the fitting coefficients of determination r² are >0.9984. The detection limits (LODs) were in the range of 0.002–0.1 μg/g. The recoveries were able to reach 82.5–103.6%, while the matrix effects ranged from 87.7 to 109.3%. The intra- and inter-day accuracies were in the range of 82.3–113.8%, while the intra- and inter-day precision ranged from 0.4 to 13.6%. Among 40 batches of health foods and 32 batches of CTPMs (including 28 capsules, 32 tablets, 10 liquid and 2 pills) samples, 28 batches of heath foods were positive. The detected chemical ingredients involved sildenafil, tadalafil, aildenafil and sulfoaildenafil. This method can be used for the screening, identification and quantification of illegal adulterated aphrodisiac chemical ingredients in health foods and CTPMs. Moreover, the LC-Q-TOF MS is very useful to structural elucidation of unknown compound.