大鼠肺中多壁碳纳米管测量的改进方法。
Improved method for measurement of multi-walled carbon nanotubes in rat lung.
作者信息
Ohnishi Makoto, Suzuki Masaaki, Yamamoto Masahiro, Kasai Tatsuya, Kano Hirokazu, Senoh Hideki, Higashikubo Ichiro, Araki Akihiro, Fukushima Shoji
机构信息
Japan Bioassay Research Center, Japan Organization of Occupational Health and Safety, 2445 Hirasawa, Hadano, Kanagawa 257-0015 Japan.
Occupational Health Research and Development Center, Japan Industrial Safety and Health Association, 5-35-1 Shiba, Minato-ku, Tokyo, 108-0014 Japan.
出版信息
J Occup Med Toxicol. 2016 Sep 15;11:44. doi: 10.1186/s12995-016-0132-7. eCollection 2016.
BACKGROUND
Previously, we have developed and reported the method of measuring multi-walled carbon nanotube (MWCNT) in the lung from rats exposed to MWCNT intratracheally. The present research was performed to improve the analytical method of MWCNT to measure multiple samples in a short period of time. For the xanalysis of MWCNTs from tissues, the existence of carbon black may interfere. Therefore, it was examined whether or not carbon black interfere the determination of MWCNT in the standard solutions. Then, MWCNTs were administered to rats and the MWCNTs were determined in the rats by the new method and the recovery rates and time for determination were calculated. The standard solutions for MWCNTs and carbon black were prepared, and the concentrations in the solutions were determined by HPLC with checking their linearity between the concentrations and signal intensities. The reproducibility of the determination was also checked.
METHODS
The concentrations of MWCMTs in the standard solutions were determined by HPLC with a fluorescent detector. Those of carbon black were also determined using the same method. The MWCNTs were administered to rats intratracheally. The MWCNTs in the lung were determined in a newly modified method including digestion of lung tissues by strong alkali solution and marking MWCNTs by benzo[ghi]perylene. The time for the determinations was recorded and the recovery rate of MWVNTs was calculated.
RESULTS
MWCNT showed linearity in a range of 0.2 to 1.0 μg/mL. In contrast, carbon black demonstrated a very low slope, showing flat pattern. Regarding the reproducibility of the analysis, the coefficient of variation was lower than 10 %. The analysis of 20 samples were completed in 1.5 h. The recovery rates of MWCNT from the lung of rats receiving intratracheal MWCNT administration were 101 to 102 %.
CONCLUSIONS
The improved method for measuring MWCNT allows an efficient MWCNT quantitation in a short period of time. Also, a small amount of MWCNTs can be measured without influence of carbon black.
背景
此前，我们已开发并报告了一种用于测量经气管内暴露于多壁碳纳米管（MWCNT）的大鼠肺内多壁碳纳米管的方法。本研究旨在改进MWCNT的分析方法，以便在短时间内测量多个样本。对于组织中MWCNT的分析，炭黑的存在可能会产生干扰。因此，研究了炭黑是否会干扰标准溶液中MWCNT的测定。然后，将MWCNT给予大鼠，并通过新方法测定大鼠体内的MWCNT，计算回收率和测定时间。制备了MWCNT和炭黑的标准溶液，并通过高效液相色谱法（HPLC）测定溶液中的浓度，同时检查浓度与信号强度之间的线性关系。还检查了测定的重现性。
方法
使用荧光检测器通过HPLC测定标准溶液中MWCNT的浓度。炭黑的浓度也采用相同方法测定。将MWCNT经气管内给予大鼠。采用一种新的改进方法测定肺内的MWCNT，该方法包括用强碱溶液消化肺组织并用苯并[ghi]苝标记MWCNT。记录测定时间并计算MWCNT的回收率。
结果
MWCNT在0.2至1.0μg/mL范围内呈线性。相比之下，炭黑的斜率非常低，呈平坦模式。关于分析的重现性，变异系数低于10%。20个样本的分析在1.5小时内完成。经气管内给予MWCNT的大鼠肺中MWCNT的回收率为101%至102%。
结论
改进后的MWCNT测量方法能够在短时间内高效地进行MWCNT定量。此外，少量MWCNT的测量不受炭黑的影响。
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