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采用 UHPLC-MS/MS 技术检测哥伦比亚东北部热带水库中的蓝藻毒素。

Detected cyanotoxins by UHPLC MS/MS technique in tropical reservoirs of northeastern Colombia.

机构信息

Grupo GDCON, Facultad de Ingeniería, Sede de Investigación Universitaria (SIU), Universidad de Antioquia, 70#, 52-21, Medellin, Colombia.

Grupo GDCON, Facultad de Ingeniería, Sede de Investigación Universitaria (SIU), Universidad de Antioquia, 70#, 52-21, Medellin, Colombia.

出版信息

Toxicon. 2019 Sep;167:38-48. doi: 10.1016/j.toxicon.2019.06.010. Epub 2019 Jun 8.

Abstract

This study focused on the detection and quantification of eight cyanotoxins in water samples in three reservoirs located in the eastern department of Antioquia, Colombia. The reservoirs are a source of water supply and hydroelectricity, and also generate economic activities in fishing and recreation. Between May 2015 and October 2016, 8 samplings were carried out at times of high temperatures, which ranged from 20 to 29 °C. This period was selected because of a significant or strong El Niño phenomenon, according to the World Meteorological Organization. For the study, 270 integrated samples were taken from the photic zone (PZ) and the surface of the reservoirs, at each of the three sampling points. The samples were analyzed by the analytical technique of ultra-high-performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC MS/MS). The quantification performed for six microcystins (MCs), a nodularin (NOD) and a cylindrospermopsin (CYN), showed positive results well above 1 μg L. In the water of the Abreo Malpaso and Peñol reservoirs, microcystin-LR (MC-LR), microcystin-YR (MC-YR) and [D-Asp,(E)-Dhb]- microcystin-RR toxins were detected at levels of considerable concentration, especially between May and September 2015, when there was no rainfall in this region. In the Playas reservoir, positive results for [D-Asp,(E)-Dhb]-MC-RR were detected from May to November 2015, with the highest concentrations being reached in dry season. The temperatures reached and the changes in climatic conditions witnessed during the monitoring period of this study were important factors in the production of cyanotoxins. This was evidenced in this work by the high concentrations of detected cyanotoxins and their absence in periods of rain, as happened from the second quarter of 2016 until the end of the study in October. This is the first study of the detection and quantification of cyanotoxins in tropical reservoirs of northeastern Colombia using the UHPLC MS/MS analytical technique, which allowed the toxins to be unequivocally detected and confirmed. A method was developed and validated, proving to be sensitive, reproducible and accurate. For each of the toxins (microcystin-LR (MC-LR), microcystin-RR (MC-RR), microcystin-YR (MC-YR), [D-Asp,(E)-Dhb]- microcystin-RR, microcystin-LW (MC-LW), microcystin-LF (MC-LF), nodularin (NOD) and cylindrospermopsina (CYN)) the correlation coefficients (R) were in a range between 0.9907 and 0.9999. Verification of the accuracy of the method was performed through a calibration curve in solvent. The recovery percentages of the accuracy and precision tests of the method for low level, medium level and high level were in a range between 64% and 115% for all the cyanotoxins. The validation of the cyanotoxin method shows that it is possible to detect them individually in natural water with a quantification limit (LOQ) of approximately 0.05 μg L.

摘要

本研究聚焦于在哥伦比亚安蒂奥基亚省东部的三个水库中检测和定量分析八种微囊藻毒素。这些水库是供水和水力发电的来源,也是渔业和娱乐等经济活动的发源地。2015 年 5 月至 2016 年 10 月期间,在高温期间(20-29°C)进行了 8 次采样。选择此时间段是因为世界气象组织认为这期间出现了显著或强烈的厄尔尼诺现象。为了进行这项研究,在三个采样点的每个点,从透光层(PZ)和水库表面采集了 270 个综合样本。样品通过超高效液相色谱-三重四极杆质谱联用(UHPLC MS/MS)分析技术进行分析。对六种微囊藻毒素(MCs)、一种节球藻毒素(NOD)和一种柱孢藻毒素(CYN)进行的定量分析显示,阳性结果远远高于 1μg/L。在 Abreo Malpaso 和 Peñol 水库的水中,检测到微囊藻毒素-LR(MC-LR)、微囊藻毒素-YR(MC-YR)和[D-Asp,(E)-Dhb]-微囊藻毒素-RR 毒素,其浓度相当高,特别是在 2015 年 5 月至 9 月期间,该地区没有降雨。在 Playas 水库中,从 2015 年 5 月至 11 月检测到[D-Asp,(E)-Dhb]-MC-RR 的阳性结果,在旱季达到最高浓度。在本研究的监测期间,达到的温度和气候条件的变化是产生微囊藻毒素的重要因素。这一点在这项工作中得到了证实,因为检测到的微囊藻毒素浓度很高,而在降雨期间则没有,这种情况从 2016 年第二季度一直持续到 10 月研究结束。这是首次使用 UHPLC MS/MS 分析技术在哥伦比亚东北部热带水库中检测和定量分析微囊藻毒素,该技术能够明确地检测和确认毒素。开发并验证了一种方法,证明其具有敏感性、重现性和准确性。对于每种毒素(微囊藻毒素-LR(MC-LR)、微囊藻毒素-RR(MC-RR)、微囊藻毒素-YR(MC-YR)、[D-Asp,(E)-Dhb]-微囊藻毒素-RR、微囊藻毒素-LW(MC-LW)、微囊藻毒素-LF(MC-LF)、节球藻毒素(NOD)和柱孢藻毒素(CYN)),相关系数(R)在 0.9907 到 0.9999 之间。通过溶剂中的校准曲线验证了该方法的准确性。该方法的低、中、高准确度和精密度测试的回收率在所有微囊藻毒素中为 64%至 115%。微囊藻毒素方法的验证表明,有可能在自然水中以约 0.05μg/L 的定量限(LOQ)单独检测到它们。

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