2-fluorenecarboxaldehyde hydrazone as a new fluorogenic reagent for the determination of acidity and its applications = 산도의 측정을 위한 새로운 형광 시약으로서 2-Fluorenecarboxal hydrazone의 개발과 그 응용

국문 초록 (Abstract)

산도의 측정을 위한 새로운 형광시약으로서 2-Fluorenecarboxaldehyde hydrazone(2-FCH) 을 합성하였다. 2-FCH 는 그 자체는 형광을 나타내지 않으나, 산에 의해서 excitation 392 nm 와 emission 447 nm 에서 강한 형광을 나타내므로 이러한 특성을 이용하여 산도의 측정에 이용하고자 하였다. 여러가지 무기산과 유기산에 대한 2-FCH 의 반응성을 조사하였고, 강산에 의해서 pH 0.60 에서 pH 3.90 까지 직선성이 성립되었다.
그 응용으로, 2-FCH 를 이용하여 사람과 쥐의 위산도를 측정하여 pH-meter 에 의한 측정치와 비교한 결과 2-FCH 방법에 의한 측정값이 pH-meter 에 의한 측정치에 비해 쥐의 경우 약 0.6-1.3 unit, 사람의 위액에서 악 0.8-1.2 unit 정도 크게 나타났다. 그러나, 이 두 방법을 통계적으로 처리한 결과 상관계수가 쥐의 경우 0.940, 사람의 경우에서 0.938 로 나타났으며, 결정계수 (r²) 가 각각에서 0.883, 0.879 로 나타난것으로 보아 두 측정법 사이에는 밀접한 관련성이 있는 것으로 보여진다.
수소이온의 활성도 측정에 있어서 2-FCH 방법을 통해 얻어진 활성도 계수를 종래의 Debye-Huckel 식에 의해 구해진 데이타와 비교한 결과, 각 무기산 또는 전해질의 영향에 있어서 서로 유사한 측정치를 얻었다. 위산에서 수소이온의 활성도 계수는 이미 보고된 바와 같이 순수한 염산에 비해 낮은 값을 보였다.
염산염을 가진 수종의 약물 정량에 있어서도 2-FCH 를 응용한 결과 각 약물에서 만족할 만한 상관계수를 얻을 수 있었다.
2-FCH 는 산도를 측정하는 새로운 형광시약으로서 좋은 결과를 보였으며, 수소이온의 활성도 계수 측정 및 염산염 약물의 간절 정량에도 응용될 수 있으리라 사료된다.

다국어 초록 (Multilingual Abstract)

2-Fluorenecarboxaldehyde hydrazone (2-FCH) was synthesized as a new fluorescent reagent for the determination of acidity. 2-FCH had no fluorescence in itself, however, strongly fluoresced under acidic condition at excitation maximum of 392 nm and emission maximum of 447 nm, respectively. 2-FCH reactivities were studied in various inorganic and organic acids. It showed linearities ranged from pH 0.60 to pH 3.90 by strong acids.
As applications of 2-FCH, the acidity of gastric juice in rats and humans was determined. In comparison with pH-measurement, acidity measured by this method showed generally increased values about 0.6 - 1.3 unit and 0.8 - 1.2 unit. in rats and humans, respectively. However, these results had statistically close correlation with that of pH-measurement and the correlation coefficients were 0.940 and 0.938 in rats and humans between two methods.
2-FCH method was also applied to measure activity coefficient of hydrogen ion and to determine drugs having acid salt form. Activity coefficients were compared to the calculated values by the Debye-Huckel equation and Harned's rule. Activity coefficients by 2-FCH method were similar to those by two methods at each concentration of strong inorganic acids. Drug analyses having acid salt form by 2-FCH method showed also good correlation coefficients.
As a result, 2-FCH was a good reagent as a new fluorescent agent for the determination of acidity and could be applied in various fields.

목차 (Table of Contents)

• List of Tables = i
• List of Figures = ii
• Scheme Legends = iii
• 국문초록 = iv
• General Introduction = 1
• Part I 2-Fluorenecarboxaldehyde hydrazone as a New Fluorescent Reagent = 9
• Introduction = 9
• Materials and Methods = 10
• 1) Apparatus and Chemicals = 10
• 2) Synthesis of 2-FCH = 12
• 3) Analytical parameters of spectrofluorometer = 12
• 4) Preparation of stock solutions = 13
• 5) Determination of acidity of inorganic acids with 2-FCH = 14
• 6) Effect of organic acids on the fluorescence intensity of 2-FCH = 14
• Results and Discussion = 16
• 1) Structure analysis of 2-FCH = 16
• 2) Determination of acidity of inorganic acids with 2-FCH = 24
• i) Determination of acidity of HCl = 24
• ii) Determination of acidity of H₂SO₄= 24
• iii) Determination of acidity of HNO₃= 24
• iv) Determination of acidity of H₃PO₄= 25
• v) Determination of acidity of H₂CO₃= 25
• 3) Effect of organic acids on the fluorescence intensity of 2-FCH = 30
• Conclusion = 33
• Part II Application of 2-FCH = 34
• Chapter 1. Spectrofluorometric determination of gastric juice acidity in rat and human = 34
• Introduction = 34
• Materials and Methods = 40
• 1) Apparatus and Chemicals = 40
• 2) Application to the determination of gastric juice acidity of rat = 41
• 3) Application to the determination of gastric juice acidity of human = 42
• Results and Discussion = 44
• 1) Application to the determination of gastric juice acidity of rat = 44
• 2) Application to the determination of gastric juice acidity of human = 49
• Chapter 2. Spectrofluorometric determination of activity coefficients of hydrogen ion = 56
• Introduction = 61
• Results and Discussion = 62
• 1) Activity coefficient of strong inorganic acids by 2-FCH method = 62
• 2) Effect of electrolytes on the activity coefficient of hydrogen ion = 62
• 3) Activity coefficient of hydrogen ion in gastric juice = 71
• Chapter 3. Quantitation of drug having acid salt form using 2-FCH = 76
• Introduction = 76
• Materials and Methods = 76
• Results and Discussion = 78
• 1) Determination of cysteine HCl = 78
• 2) Determination of flupentixol 2HCl = 78
• 3) Determination of zipeprol 2HCl = 78
• 4) Determination of pirenzepine 2HCl = 79
• 5) Determination of thiamine HCl = 79
• 6) Determination of cyclizine HCl = 79
• 7) Determination of melitracene HCl = 80
• Conclusion = 86
• Literature Cited = 89
• Abstract = 105
• Acknowledgement