【作者】 姜瑶；

【导师】 顾景凯；

【作者基本信息】 吉林大学 ， 微生物与生化药学， 2007， 硕士

【摘要】 本文建立了应用液相色谱-质谱联用技术(LC-MS/MS)测定血浆中奥曲肽浓度的定量方法,并采用此法测定了速释奥曲肽制剂单次给药,以及进口(瑞士诺华制药有限公司)和国产(长春金赛制药股份有限公司)两种奥曲肽控释微球制剂单次和多次给药后血浆中奥曲肽的浓度。根据结果绘制出血药浓度-时间曲线,采用梯形法计算AUC值,进行药代动力学研究;将两种控释微球制剂单次给药后AUC值进行对数转换后运用双单侧t检验分析,比较两种控释微球制剂的差异。结果显示,与短效奥曲肽比较,奥曲肽控释微球制剂能够缓慢释放奥曲肽,显著的增加奥曲肽的体内滞留时间,使其体内浓度能更长时间保持于有效浓度以上,从而达到延长给药间隔时间,减少病人痛苦的目的,有利于消化道疾病、多种内分泌肿瘤和肢端肥大症的治疗。经过双单侧t检验法分析测得的数据,结果表明金赛和诺华生产的两种控释微球制剂没有显著性差异,多次用药后在体内也并无蓄积作用。更多还原

【Abstract】 AIM: Develop a rapid and sensitive method for the determination of octreotide in beagle dog’s plasma by liquid chromatography-mass spectrometry (LC-MS/MS). For the pharmacokinetics study, the method were used to quantify the octreotide concentrations in plasma after an intravenous injection of sandostatin and intramuscular injections of sandostatin LAR and Jinsai octreotide LAR.STUDY METHOD: Octreotide and the internal standard, triptorelin, were precipitated from the matrix by acetonitrile, then washed with dichloromethane and separated on a Zorbax-extend C18 column employing a 1% formic acid/methanol gradient system. Detection was carried out by multiple reaction monitoring (MRM) on an API 4000 LC-MS/MS system with an ESI interface in the positive ion mode. MRM was performed at low resolution using the mass transition ion-pairs m/z 510.5→m/z 120.1 and m/z 656.5→m/z 175.8 for octreotide and triptorelin, respectively. The linearity, specificity, precision, accuracy, recovery and stability were estimated for the validations of the assay.The concentrations of octreotide in health beagle dogs’plasma were measured after intravenous injection of sandostatin at 0.022 mg/kg and intramuscular injections of sandostatin LAR and Jinsai octreotide LAR (1.4 mg/kg). After the plasma concentration–time profile was shown, the area under the curve (AUC0-t) was determined by the linear trapezoidal rule. The two one-sided tests procedure carried out at a significance level of 0.05, and the mean Jinsai octreotide LAR/sandostatin LAR ratios of AUC0?t (after log-transformed) were computed.RESULTS: A highly selective, sensitive, rapid and reproducibility method for the determination of octreotide in beagle dogs’plasma by using LC-MS/MS has been developed and validated in this study. The standard curve of octreotide was linear over a working range of 0.050-50 ng/ml with the lower limit of quantitation (LLOQ) of 0.050 ng/ml and the limit of detection (LOD) of 0.020 ng/ ml. The accuracy was in the range 90.1-109% and the intra- and inter-day precision was <8.37%. It showed relative high and stable recovery for the sample preparing procedure. The samples were stable in three freeze-thaw cycles test, in the autosampler at room temperature for 12 h and in storage at ?20°C for 60 days. The methods, which kept the conformance to the relevant standards of Pharmacopoeia of People’s Republic of China, were ideally suited for the study of pharmacokinetics of octreotide.The results of pharmacokinetics study of octreotide showed that: Compared to sandostatin, the LAR octreotide, which required only one monthly intramuscular injection depending on its sustained release, avoided the repeated daily hypodermic injections or prolonged intravenous infusions and exhibited wider application in the clinical treatment of disease of digestive tract, endocrine tumors and acromegaly. Furthermore, both sandostatin LAR and Jinsai octreotide LAR had no cumulative effect after repeated intramuscular injections.The results of bioequivalence evaluation of octreotide LAR showed that: The mean AUC0-t (0-1200 h) of sandostatin LAR after intramuscular injected at dose of 1.4 mg/kg was 578.9±83.04 ng·h/ml, while that of Jinsai octreotide LAR with the same administration dose was 568.0±256.1 ng·h/ml. The relative bioavailability of Jinsai octreotide LAR (sandostatin LAR as reference) was 98.12%. The two one-sided tests illustrated that there was no significant difference in bioavailability of the two LAR microsphere formulations.更多还原