理科教育学研究

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1 原著論文 Vol.No. doi:. /sjst. レーザー光を利用した簡易屈折計によるグリセリンの定量リパーゼによる油脂の分解反応の測定田中謙介 Candida rugosa CRL CRL 1 [ キーワード ] 1. はじめに Rogalska, E., Cudrey, C., Ferrato, F., Verger, R., C. rugosa CRL Rogalska, E. et al., 1 兵庫県立神戸鈴蘭台高等学校

2 2. グリセリンの定量 2.1 グリセリン溶液の濃度と屈折率の関係 n 1 N n1 1 N n2 1 1 x x n 2 N n 2 N n x n n n N N N C N A CN A xn n 1 N n1 1 1 n2 1 1 n CN n 2 N1 n1 2 N2 n2 2 N1 n1 2 n n 2 n2 1 6n 1 2 n2 2 n 3 n n N n1 1 1 n2 1 1 n CN N1 n1 2 N2 n2 2 N1 n1 2 N N C n D Na D. nm A A 2.2 簡易屈折計の原理 sin 1 n2 sin 3 n sin n sin n n n n tan tan 図 2 装置の原理図 1 グリセリン水溶液の屈折率 n D (20 C) 図 3 入射角 θ 1 と tan θ 4 の関係

3 Vol.No. 図 5 簡易屈折計の概略図図 4 グリセリン水溶液の屈折率 n 2 と tan θ 4 の関係 n D. n D. tan L 図 6 簡易屈折計 ( 写真 ) 2.3 簡易屈折計の製作 mm mm mm mm mm RAY nm mw FujiCorona ML- nm max. mw m nm mm nm mm mm 2.4 グリセリン溶液の検量線.. ph. Na HPO. g L A KH PO. g L B A B.. ph.. g L C. g L D C D..

4 ... g ph. L. mol/l ph..... mol/l ph.. mol/l.. nm nm -... mol/l 2.5 グリセリン定量のための基礎実験.. ph. C. mol/l 図 7 グリセリン溶液濃度と光点の移動距離の関係表 1 溶媒ごとの検量線の傾き mm/(moll - ) ph ph.... S.D... - ph. ph. ml ml C rpm ml.. CRL L - G, SIGMA-ALDRICH. g ph. ml

5 Vol.No. 図 8 移動距離の温度依存性 ph. λ= nm 表 2 緩衝液 +オリーブ油における屈折計測定値の変動 hrs..... mm..... C rpm ml mm. g ph. ml 油脂のリパーゼによる分解反応 3.1 実験方法 ph. ml CRL. g ml ml C rpm C ml. ml. ml 3.2 滴定法による脂肪酸の定量 JIS K... Vol% %. mol/l. W/V%... ml ml. mol/ L NaOH

6 3.3 屈折法によるグリセリンの定量..... mol/l ph.... μm ADVANTEC CSAN LP ITALIANA SPA ml 3.4 実験結果と考察 % % rpm 図 11 C. rugosa によるオリーブ油の分解図 9 分解反応の装置概略図 10 自作の防水型マグネチックスターラー図 12 グリセリンと脂肪酸の物質量比 S.D.

7 Vol.No.. S.D.... S.D.. mm 3.5 屈折法と酵素法の比較 JK F 消化酵素入り胃腸薬を用いた実験 AP mg/ CRL.. AP mg ml ADVANTEC mm ml ph... ml. - mol CRL... AP u/g CRL u/mg CRL 4. 部活動における実践 ph L CRL 表 3 屈折法と酵素法の比較 mol/l / h S.D..

8 図 13 豚膵臓リパーゼによるオリーブ油の分解図 15 CRL の至適 ph 図 14 CRL によるオリーブ油の分解 - - Okumura, S., Iwai, M., Tsujisaka, Y., CRL ph ph C C 5. おわりに図 16 CRL の至適温度謝辞註 screw-top KEEPER ml

Vol.No. 防水型マグネチックスターラーの設計図 No. NT. V rpm L- OIL IFU OIV HP Reference; OIV-MA-AS-Retrieved from http://www.oiv.int/oiv/info/enmethodesinternationalesvin #autres nm Retrieved from http://www.

9 Vol.No. 防水型マグネチックスターラーの設計図 No. NT. V rpm L- OIL IFU OIV HP Reference; OIV-MA-AS-Retrieved from #autres nm Retrieved from AP HP &wc= 引用文献... JIS K Retrieved from com/k /K - -.html Okumura, S., Iwai, M., Tsujisaka, Y. ( ). Positional Specificities of Four Kinds of Microbial Lipases. Agr. Biol. Chem., 40( ),. Rogalska, E., Cudrey, C., Ferrato, F., Verger, R. ( ). Stereo selective Hydrolysis of Triglycerides by Animal and Microbial lipases. Chirality, 5,. Retrieved from ed.jp/science/ronnbunshu/top.htm. One Point. Retrieved from utsunomiya-u.ac.jp/v n /yoshida / Yoshida, A. ( ). A Simple Experimental Teaching Material for Understanding Digestion of Lipids with Lipase.. Retrieved from n /yoshida /..

10 Quantitative Analysis of Glycerol by the Use of a Handmade Refractometer Measuring the Decomposition of Oils by Lipases Kensuke TANAKA Kobe Suzurandai Senior High School, Hyogo SUMMARY This paper describes the development of an experimental device to measure the glycerol produced from the decomposition of oils and fats by lipases. This device is made from three slide glasses formed into a prism shape. A solution is then poured into this prism and a laser beam is irradiated from the side. The position of the laser beam on the screen moves in proportion to the concentration of the solution. Lipase from Candida rugosa (CRL) is utilized for this experiment, because of its ability to decompose oils and fats into fatty acids and glycerol. The difference between the measurements by this method and by those measurements recorded by the enzyme method, which is the current official method of measurement recorded, was 7% of the average. The molarity of glycerol showed approximately 1/3 fatty acids after 3 hours reaction. Subsequently, I introduced this new device to the science club. The theme of the activity was to examine the positional specificity difference between CRL and porcine pancreatic lipase (PPL) which decomposes oils to fatty acids and monoglycerides. The results of the students experiments showed no remarkable increase in the glycerol content when using PPL as opposed to CRL. This device can be easily utilized by students, and as such, has the potential to become an effective tool for future educational activities. <Key words> Refractometer, laser beam, Glycerol, Quantitative analysis, Lipase

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