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采用液相色谱-串联质谱法(LC-MS/MS)在克氏-亨氏溶液中测定6-氰基多巴胺、6-硝基多巴、6-硝基多巴胺和6-硝基肾上腺素。评估兔离体右心房和心室的基础释放量。

Measurement of 6-cyanodopamine, 6-nitrodopa, 6-nitrodopamine and 6-nitroadrenaline by LC-MS/MS in Krebs-Henseleit solution. Assessment of basal release from rabbit isolated right atrium and ventricles.

作者信息

Júnior Gilberto Quirino, Britto-Júnior José, Magalhaes Tainah Babadopulos, Campos Rafael, Nyamkondiwa Kudzai L, Klugh Kameron L, Peterson Larryn W, Corvino Angela, Sparaco Rosa, Frecentese Francesco, Caliendo Giuseppe, De Nucci Gilberto

机构信息

Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.

Department of Chemistry, Rhodes College, Memphis, Tennessee, USA.

出版信息

Biomed Chromatogr. 2023 Sep;37(9):e5691. doi: 10.1002/bmc.5691. Epub 2023 Jul 10.

Abstract

This study presents the validation of a sensitive method for the determination of 6-nitrodopa, 6-nitrodopamine, 6-nitroadrenaline and 6-cyanodopamine in Krebs-Henseleit solution by LC-MS/MS with ESI . HRMS was used to precisely characterize the structures of the fragment ions. The method was applied to investigate the catecholamine basal release from rabbit isolated atria and ventricles. The atria and ventricles were suspended separately in a 5 ml organ bath containing Krebs-Henseleit solution with ascorbic acid (3 mM), gassed (95%O /5%CO ) at 37°C for 30 min. Strata-X 33 μm SPE cartridges were employed for the extraction of the catecholamines and the internal standard 6-nitrodopamine-d . The catecholamines were separated employing a 150 × 3 mm Shim-pack GIST C18-AQ (3 mm particle size), placed in an oven at 40°C and perfused by 65% of mobile phase A (MeCN/H O; 90/10, v/v) + 0.4% CH COOH and 35% mobile phase B (deionized H O) + 0.2% CH O at 320 μl/min in isocratic mode. The method was linear at 0.1-20 ng/ml. The method was used to identify for the first-time basal release of the three nitrocatecholamines mentioned above and a member of a novel class of catecholamines, the cyanocatecholamines.

摘要

本研究介绍了一种灵敏方法的验证过程,该方法用于通过电喷雾电离液相色谱-串联质谱法(LC-MS/MS)测定Krebs-Henseleit溶液中的6-硝基多巴、6-硝基多巴胺、6-硝基肾上腺素和6-氰基多巴胺。采用高分辨率质谱(HRMS)精确表征碎片离子的结构。该方法用于研究兔离体心房和心室中儿茶酚胺的基础释放。将心房和心室分别悬挂于含有抗坏血酸(3 mM)的5 ml Krebs-Henseleit溶液器官浴中,在37°C下用(95%O₂/5%CO₂)通气30分钟。采用Strata-X 33μm固相萃取柱提取儿茶酚胺和内标6-硝基多巴胺-d₅。使用150×3 mm Shim-pack GIST C18-AQ(3μm粒径)分离儿茶酚胺,该色谱柱置于40°C的柱温箱中,以等度模式,用65%的流动相A(乙腈/水;90/10,v/v)+0.4%乙酸和35%的流动相B(去离子水)+0.2%乙酸,以320μl/min的流速进行洗脱。该方法在0.1-20 ng/ml范围内呈线性。该方法首次用于鉴定上述三种硝基儿茶酚胺以及一类新型儿茶酚胺——氰基儿茶酚胺的基础释放。

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