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用于测定白花丹素以诊断白花丹中毒的薄层色谱法和高效液相色谱法

TLC and HPLC methods for the determination of plumbagin for the diagnosis of poisoning by Plumbago scandens L.

作者信息

Teixeira Carlos José Rocha, Dos Santos Barbara Pereira, Saraiva Carolina Julia Costa, Pedroza Heloísa de Paula, Oloris Silvia Catarina Salgado, Soto-Blanco Benito

机构信息

Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais (UFMG), Avenida Presidente Antônio Carlos 6627, Belo Horizonte, MG, 31275-013, Brazil.

Centro Universitário Presidente Antônio Carlos (UNIPAC), Rodovia MG 482, Km 3, Conselheiro Lafaiete, MG, Brazil.

出版信息

Toxicon. 2024 Feb 23;239:107634. doi: 10.1016/j.toxicon.2024.107634. Epub 2024 Feb 1.

Abstract

Plumbago scandens L. (Plumbaginaceae) occurs in all regions of Brazil. It has been described as toxic to cattle and goats. Caustic lesions in the upper digestive tract characterize poisoning. P. scandens contains a naphthoquinone named plumbagin, which presents high cytotoxic activity. Plumbago auriculata Lam., a widely used ornamental plant, is considered potentially toxic, but there is limited data about its toxicity. This work aimed to validate analytical methodologies for determining the levels of plumbagin in samples of leaves, stems, and rumen content to be used as an auxiliary chemical marker in the laboratory diagnosis of intoxication. One methodology used thin layer chromatography (TLC), and another used high-performance liquid chromatography (HPLC). The presence of palisade grass (Urochloa brizantha (Hochst. ex A.Rich.) R.D.Webster), Guinea grass (Megathyrsus maximus (Jacq.) B.K.Simon & S.W.L.Jacobs), corn silage, and rumen content did not interfere with plumbagin in the two methodologies. The TLC methodology generates qualitative results but is simple to implement and has a low cost. The HPLC methodology showed a limit of detection (LOD) of 0.01 μg/mL and a limit of quantification (LOQ) of 0.05 μg/mL. Leaf and stem samples of P. scandens evaluated showed high levels of plumbagin (0.261 ± 0.087 % and 0.327 ± 0.055 %, respectively). In contrast, leaves of P. auriculata did not show detectable levels of the toxin, and some stem samples showed low levels (up to 0.000114 %). Thus, these methodologies can be used to confirm or rule out the consumption of P. scandens in rumen content from animals suspected of poisoning.

摘要

攀缘白花丹(白花丹科)分布于巴西的所有地区。它被描述为对牛和山羊有毒。上消化道的腐蚀性损伤是中毒的特征。攀缘白花丹含有一种名为白花丹醌的萘醌,具有很高的细胞毒性活性。蓝花丹是一种广泛使用的观赏植物,被认为具有潜在毒性,但关于其毒性的数据有限。这项工作旨在验证用于测定叶片、茎和瘤胃内容物样本中白花丹醌含量的分析方法,以作为中毒实验室诊断中的辅助化学标志物。一种方法使用薄层色谱法(TLC),另一种方法使用高效液相色谱法(HPLC)。在这两种方法中,栅状臂形草(臂形草)、大盖草(巨穗苔草)、玉米青贮饲料和瘤胃内容物的存在均不干扰白花丹醌的测定。TLC方法产生定性结果,但易于实施且成本低。HPLC方法的检测限(LOD)为0.01μg/mL,定量限(LOQ)为0.05μg/mL。所评估的攀缘白花丹叶片和茎样本显示出高水平的白花丹醌(分别为0.261±0.087%和0.327±0.055%)。相比之下,蓝花丹的叶片未显示出可检测到的毒素水平,一些茎样本显示出低水平(高达0.000114%)。因此,这些方法可用于确认或排除疑似中毒动物瘤胃内容物中是否摄入了攀缘白花丹。

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