UPLC-MS/QAMS测定小鼠血浆中石斛碱及其代谢产物M-250、M-280的浓度
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篇名: UPLC-MS/QAMS测定小鼠血浆中石斛碱及其代谢产物M-250、M-280的浓度
TITLE:
摘要: 目的:首次建立测定小鼠血浆中石斛碱及其代谢产物M-250、M-280浓度的方法。方法:小鼠灌胃石斛碱60 mg/kg,1 h后收集血浆处理后,以盐酸伪麻黄碱为内标,以石斛碱对照品为对照,采用超高效液相色谱-质谱/一测多评法测定石斛碱及其代谢产物M-250、M-280的血药浓度。色谱柱为Hypersil Gold C18,流动相为0.1%甲酸水溶液-乙腈(梯度洗脱),流速为0.3 mL/min,柱温为40 ℃,进样量为5 μL;采用可加热电喷雾离子源,全扫描/正离子模式,雾化温度为300 ℃,离子传输管温度为350 ℃,鞘气流速为35 arb,辅助气流速为15 arb,喷雾电压为3.5 kV,碰撞电压为30、40、50 eV,质荷比检测范围为100~1 500。结果:小鼠血浆中内源性物质对石斛碱及其代谢产物M-250、M-280浓度测定无干扰;石斛碱检测质量浓度线性范围为9.13~912.94 ng/mL(r=0.999 6);定量下限为3.04 ng/mL,日内、日间精密度的RSD≤7.5%(n=5或n=3);准确度为96.8%~107.5%(n=5);基质效应为97.1%~106.0%(RSD=1.8%~4.7%,n=5);质控血浆样品溶液15 ℃保存24 h、-70 ℃保存并反复冻融3次以及-70 ℃保存15 d后浓度的RSD≤12.8%(n=3)。小鼠血浆样品中石斛碱质量浓度为(41.3±5.7) ng/mL(n=12),以相对校正因子(RCF)按1.0计,代谢产物M-250、M-280的质量浓度分别为(493.0±73.1)、(41.4±3.0) ng/mL(n=12)。结论:该方法灵敏、准确,可用于小鼠血浆中石斛碱及其代谢产物M-250、M-280的浓度测定。
ABSTRACT: OBJECTIVE: To establish a method for the determination of dendrobine and its metabolites M-250 and M-280 in mice plasma for the first time. METHODS: Mice were given dendrobine 60 mg/kg by intragastric administration, 1 h later plasma were collected and treated. Using pseudoephedrine hydrochloride as internal standard and dendrobine reference substance as control, the plasma concentrations of dendrobine and its metabolites M-250 and M-280 were determined by UPLC-MS combined with quantitative analysis of multi-components by single marker. The separation was performed on Hypersil Gold C18 column with 0.1% formic acid-acetonitrile (gradient elution) at the flow rate of 0.3 mL/min. The column temperature was set at 40℃, and sample size was 5 μL. Heatable electrospray ionization (HESI) source, scan/ESI+ were applied and operated in positive ion mode with atomization temperature of 300 ℃, ion transmission tube temperature of 350 ℃, the sheath gas velocity of 35 arb, the auxiliary air velocity of 15 arb, the spray voltage of 3.5 kV, the collision voltage of 30, 40, 50 eV. The mass-to-charge ratio of detection range were 100-1 500. RESULTS: The endogenous substances of mice plasma had no interference with the content determination of dendrobine and its metabolites M-250 and M-280. The linear range of dendrobine were 9.13-912.94 ng/mL (r=0.999 6). The limit of quantitation was 3.04 ng/mL. RSDs of intra-day and inter-day were all less than 7.5% (n=5 or n=3). The accuracy were 96.8%-107.5% (n=5). Matrix effects were 97.1%-106.0%(RSD=1.8%-4.7%,n=5). RSDs of the content of sample at 15℃ for 24 h, at -70 ℃ after three times freeze-thaw, at -70 ℃ for 15 d were lower than 12.8% (n=3). The content of dendrobine in plasma sample of mice was (41.3±5.7) ng/mL(n=12). The contents of its metabolites M-250 and M-280 were (493.0±73.1) and (41.4±3.0) ng/mL (n=12) with Relative correction factor of 1.0. CONCLUSIONS: The method is sensitive and accurate, and can be used for content determination of dendrobine and its metabolites M-250 and M-280 in mice plasma.
期刊: 2018年第29卷第11期
作者: 鲁艳柳,黄思,刘浩,曾瑶,王建秋,徐亚沙,陆远富,何芋岐
AUTHORS: LU Yanliu,HUANG Si,LIU Hao,ZENG Yao,WANG Jianqiu,XU Yasha,LU Yuanfu,HE Yuqi
关键字: 石斛碱;代谢产物;M-250;M-280;小鼠;超高效液相色谱-质谱法;一测多评法
KEYWORDS: Dendrobine; Metabolites; M-250;M-280; Mice; UPLC-MS; Quantitative analysis of multi-components with single marker
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