LC-UV/MS methods for the analysis of prochelator-boronyl salicylaldehyde isonicotinoyl hydrazone (BSIH) and its active chelator salicylaldehyde isonicotinoyl hydrazone (SIH).
LC-UV/MS methods for the analysis of prochelator-boronyl salicylaldehyde isonicotinoyl hydrazone (BSIH) and its active chelator salicylaldehyde isonicotinoyl hydrazone (SIH).
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DOI：
10.1016/j.jpba.2014.11.044
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发表时间：
2015-02
影响因子：
3.4
通讯作者：
Kovarikova, Petra
中科院分区：
文献类型：
--
作者：
Bures, Jan;Jansova, Hana;Stariat, Jan;Filipsky, Tomas;Mladenka, Premysl;Simunek, Tomas;Kucera, Radim;Klimes, Jiri;Wang, Qin;Franz, Katherine J.;Kovarikova, Petra
关键词：
水杨醛异烟酰腙（SIH）是一种细胞内铁螯合剂，在体外和体内都具有良好的抗氧化损伤作用。然而，其具有由腙键的快速水解引起的短生物半衰期。最近，硼酸盐prochelators的概念已被引入作为一种策略，可能会克服这些限制。本研究提出了两种互补的分析方法，用于检测前螯合剂BSIH（硼酰水杨醛异烟酰腙）沿着其活性金属结合螯合剂SIH在不同的溶液基质和浓度范围。分别在7 - 115 μ M和4 - 115 μM浓度范围内验证了用于测定缓冲液和细胞培养基中BSIH和SIH的LC-UV方法，并将其应用于体外BSIH活化实验。验证了LC-MS测定法，用于分别在0.06 - 23和0.24 - 23 μM浓度范围内定量血浆中的BSIH和SIH，并将其应用于体外血浆稳定性研究以及大鼠静脉注射BSIH后采集的血浆分析。Zorbax-RP bonus色谱柱和含有EDTA磷酸盐缓冲液或甲酸铵和甲醇/乙腈混合物的移动的相分别为LC-UV和LC-MS分析提供了合适的条件。在分析之前，用甲醇稀释或沉淀样品。这些分离分析技术建立了第一个经验证的方案，以研究多种基质中过氧化氢对BSIH的激活作用，直接比较前螯合剂和螯合剂在血浆中的稳定性，并提供该前螯合剂的第一个基本药代动力学数据。实验表明，BSIH是稳定的，在所有测试的媒体和部分转化为SIH由H2 O2。从体内研究的血浆样品中观察到的BSIH的完整性表明，前螯合的概念可能是一个有前途的战略，进一步发展的芳酰腙细胞保护剂。
Salicylaldehyde isonicotinoyl hydrazone (SIH) is an intracellular iron chelator with well documented potential to protect against oxidative injury both in vitro and in vivo. However, it suffers from short biological half-life caused by fast hydrolysis of the hydrazone bond. Recently, a concept of boronate prochelators has been introduced as a strategy that might overcome these limitations. This study presents two complementary analytical methods for detecting the prochelator BSIH (boronyl salicylaldehyde isonicotinoyl hydrazone) along with its active metal-binding chelator SIH in different solution matrices and concentration ranges. An LC-UV method for determination of BSIH and SIH in buffer and cell culture medium was validated over concentrations of 7 – 115 and 4 – 115 μM, respectively, and applied to BSIH activation experiments in vitro. An LC-MS assay was validated for quantification of BSIH and SIH in plasma over the concentration range of 0.06 – 23 and 0.24 – 23 μM, respectively, and applied to stability studies in plasma in vitro as well as analysis of plasma taken after i.v. administration of BSIH to rats. A Zorbax-RP bonus column and mobile phases containing either phosphate buffer with EDTA or ammonium formate and methanol/acetonitrile mixture provided suitable conditions for the LC-UV and LC-MS analysis, respectively. Samples were diluted or precipitated with methanol prior to analysis. These separative analytical techniques establish the first validated protocols to investigate BSIH activation by hydrogen peroxide in multiple matrices, directly compare the stabilities of the prochelator and chelator in plasma, and provide the first basic pharmacokinetic data of this prochelator. Experiments reveal that BSIH is stable in all media tested and is partially converted to SIH by H2O2. The observed integrity of BSIH in plasma samples from the in vivo study suggest that the concept of prochelation might be a promising strategy for further development of aroylhydrazone cytoprotective agents.
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影响因子：
15
作者：
Charkoudian, Louise K.;Pham, David M.;Franz, Katherine J.
通讯作者：
Franz, Katherine J.
影响因子：
4.1
作者：
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影响因子：
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Vávrová, K
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作者：
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