Improvement of operational efficiency in the indophenol method and quantification of ammonia excreted from fish in school class

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  • インドフェノール法の作業効率化と授業における魚類から排出されたアンモニアの定量
  • インドフェノール法の作業効率化と授業における魚類から排出されたアンモニアの定量 : マイクロプレートリーダーの教育実験機器としての可能性
  • インドフェノールホウ ノ サギョウ コウリツカ ト ジュギョウ ニ オケル ギョルイ カラ ハイシュツ サレタ アンモニア ノ テイリョウ : マイクロプレートリーダー ノ キョウイク ジッケン キキ ト シテ ノ カノウセイ
  • Potential of microplate reader as educational instrument for data analysis
  • ―マイクロプレートリーダーの教育実験機器としての可能性―

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Abstract

<p>In biological experiments in schools, students need to acquire a large amount of numerical data in science class in order to perform graphing and statistical analysis. Microplate reader, a kind of the spectrophotometers used in various research such as drug discovery and physiology, can measure light absorbance of sample solutions in microplate at once. The most common microplate format used in academic research laboratories or clinical diagnostic laboratories is 96 wells with a typical reaction volume between 100 ~ 200 μL per well. As an educational experiment to realize physiology and ecology, a method for determining the ammonia concentration by colorimetry has already been established. However, since the density is determined based on the color chart, it is difficult to perform a numerical analysis. Therefore, we developed ammonia assay using the microplate leader by indophenol method. In particular, the volume of reagents and samples was unified at 200 μL to make it easier for students to handle. We found that, light absorbance in 650 nm, an absorptivity wavelength of indophenol blue, reached the peak when sodium hydroxide and effective chlorine of sodium hypochlorite were 125 mM and 0.0065% at the final concentration. We next performed a student experiment based on this result, all the students could make standard curve and determine 1 ~ 3 ppm ammonia in water kept goldfish. Since the microplate reader can measure the light absorbance of 96 samples in approximately one minute, it will be an effective instrument to realize inquisitive biological experiments using numerical values.</p>

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