流动注射化学发光法测定激素类药物的研究及应用
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摘要
激素类药物就是以人体或动物激素(包括与激素结构、作用原理相同的有机物)为有效成分的有机物。激素类药物可以分为:糖皮质激素、肾上腺皮质激素、去甲肾上腺激素、孕激素、雌激素、雄激素等。该类药物常见使用方式有静脉使用、口服、外用及其他。常见剂型有注射粉针、水针、胶囊、片剂、霜剂、膏剂、气雾剂。激素类药物可以用来预防、治疗、诊断疾病和帮助机体恢复正常机能。例如:糖皮质激素类主要影响糖和蛋白质等代谢,且能对抗炎症反应;肾上腺激素类药物具有抗炎、抗过敏作用,临床上用于各种严重的过敏性疾病、严重的支气管哮喘、血小板减少性紫癜、剥脱性皮炎、神经性皮炎、湿疹等。
化学发光(Chemiluminescence,CL),具有灵敏度高、线性范围宽、设备简单、操作方便、分析速度快、易于实现自动化等优点,在环境、药物分析和医学研究方面具有独特的优势。将流动注射化学发光用于药物分析不仅具有理论意义,而且还有重要的实用价值。
本文共分为两个部分。
第一部分(第一章)介绍流动注射化学发光法的应用及特点;详细介绍了几种常见的化学发光体系在药物分析中的研究应用;阐述了本章的研究内容和特点。
第二部分是研究报告(第二～六章)。
论文的第二章,研究了在碱性条件下,醋酸泼尼松能降低NBS-钙黄绿素的化学发光,其强度与醋酸泼尼松的质量浓度在一定范围内成线性关系。建立了测定醋酸泼尼松的流动注射化学发光新方法,并详细研究了影响化学发光强度的因素。在优化实验条件下,醋酸泼尼松的线性范围为2.0×10-8～1.0×10-5 g/mL,检出限(3σ)为1.3×10-8 g/mL。本法成功应用于片剂中醋酸泼尼松的测定,并对该体系的机理进行探讨(已被Analytical Letters录用)。
论文的第三章研究了在碱性条件下,雌二醇对N-溴代丁二酰亚胺(NBS)-钙黄绿素化学发光体系的化学发光具有强烈的抑制作用,其强度与雌二醇的质量浓度在一定范围内成线性关系。基于此首次建立了雌二醇的化学发光抑制分析法,并详细研究了影响化学发光强度的因素,对抑制化学发光可能的机理进行了探讨。方法对雌二醇响应的线性范围为2.0×10-8～8.0×10-6 g/mL,检出限(3σ)为8.5×10-9 g/mL,对1.0×10-7 g/mL的雌二醇进行了11次平行测定,相对标准偏差为1.2 %。方法应用于注射液中雌二醇含量的测定,结果令人满意(已在分析测试学报发表)。
论文的第四章基于在硫酸介质中,苯甲酸雌二醇可以增强KMnO4-邻菲咯啉钌的化学发光反应,进一步研究发现,Na2SO3的加入,大大增强KMnO4-邻菲咯啉钌-苯甲酸雌二醇体系的发光强度。建立了一种灵敏、快速、选择性好的测定苯甲酸雌二醇的流动注射化学发光新方法。在优化的条件下,该法线性范围ΔI=2914+11.71×108c (r=0.9961),检出限为2.4×10-8 g/mL。本法成功应用于苯甲酸雌二醇针剂的测定,并对该体系机理进行探讨(已被Luminescence录用)。
论文的第五章研究了在酸性条件下,高锰酸钾能够氧化黄体酮从而产生微弱的化学发光信号,当该反应体系中存在亚硫酸钠时,体系发光信号显著增强。建立了一种灵敏、简单、快速测定黄体酮的化学发光新方法。在优化的条件下,黄体酮的线性范围为8.0×10-8～1.0×10-6 g/mL,检出限(3σ)为4.6×10-8 g/mL。该方法已成功应用于合成样品中黄体酮含量的测定。
论文的第六章,在H2SO4介质中,酸性高锰酸钾溶液可以氧化雌二醇从而产生微弱的化学发光信号,当该反应体系中存在罗丹明B时,体系发光信号显著增强,据此建立了一种简单、快速测定雌二醇的流动注射化学发光新方法。基于这一现象,本文建立了一种化学发光测定雌二醇的新方法。该方法的线性范围为2.0×10-8～1.0×10-5 g/mL,检出限(3σ)为1.2×10-8 g/mL (3σ)。该方法已成功应用于注射液中雌二醇含量的测定。
Hormones are component of organic matters which are effective for human or animals. Hormone drugs include: glucocorticoid, adrenocorticotropic, norepinephrine, progesterone, estrogen, androgen, et al. Their common usage are intravenous, oral, topical and others. Common dosage forms include injection of powder, water injection, capsules, tablets, creams, pastes, aerosols. Hormones can be used to prevent, treat, diagnose disease and help the body return to normal function. For example, glucocorticoids mainly affects sugar and protein metabolism, and can fight inflammation.
Due to its high sensitivity, simplicity and relatively low cost, chemiluminescence analysis has been widely used in biological engineering, molecular biologic, medicine, environmental chemistry.
The thesis consists of two parts.
The first one is a brief review dealing with the research and development of chemiluminescence. The other is research reports.
In Chapter two, a nivel flow injection chemiluminescence method for the determination of prednisone acetate was developed. This method is based on that the strong CL of N-bromosuccinimide (NBS) and calcein can be inhibited by prednisone acetate. The CL intensity correlated linearly with the concentration of prednisone acetate over the range of 2.0×10-8~1.0×10-5 g/mL, the detection limit (3σ) of prednisone acetate was 1.3×10-8 g/mL. This method was applied to the determination of prednisone acetate in the tablets with good results. The mechanisms of CL reaction were also studied.
In Chapter three, a nivel flow injection chemiluminescence method for the determination of estradiol was developed. This method is based on that the strong CL of N-bromosuccinimide (NBS) and calcein can be inhibited by estradiol. The CL intensity correlated linearly with the concentration of estradiol over the ranges of 2.0×10-8~8.0×10-6 g/mL. The detection limit (3σ) of estradiol was 8.5×10-9 g/mL. This method was applied to the determination of estradiol in the injections. The mechanisms of CL reaction were also studied.
In Chapter four, a novel chemiluminescence (CL) method for the fast, simple and sensitive determination of estrogen benzoate using the reaction of tris (1,10-phenanthroline) ruthenium (II)-Na2SO3-permanganate is described. This method is based on the CL reaction of estrogen benzoate (EB) with acidic potassium permanganate and tris (1,10-phenanthroline) ruthenium (II). The CL intensity is greatly enhanced when Na2SO3 is added. After optimization of the different experimental parameters, a calibration graph for estrogen benzoate is linear in the range of 5.0×10-8~1.0×10-5 g/mL. The limits (3σ) of detection is 2.4×10-8 g/mL and the relative standard deviation was 1.3 % for 1.0×10-6 g/mL estrogen benzoate (n = 11). This proposed method was successfully applied to commercial injection samples. The mechanism of CL reaction was also studied.
In Chapter five, a new chemiluminescence method is proposed for the determination of progesterone, which is based on the reaction of KMnO4- NaSO3 progesterone system. Under the optimum conditions, the linear range of progesterone is 8.0×10-8~1.0×10-6 g/mL. The detection limit (3σ) is 4.6×10-8 g/mL for progesterone.
In Chapter six, to describe a novel flow injection chemiluminescence method for the determination of estradiol. The method is based on the oxidation of estradiol by acidic permanganate solution to give weak CL signal, which is dramatically enhanced by rhodamine B. The CL signal is linear with estradiol concentration in the range 2.0×10-8~1.0×10-5 g/mL with a detection limit of 1.2×10-8 g/mL (3σ). The method has been applied to the determination of estradiol in pharmaceutical preparations.
化学发光(Chemiluminescence,CL),具有灵敏度高、线性范围宽、设备简单、操作方便、分析速度快、易于实现自动化等优点,在环境、药物分析和医学研究方面具有独特的优势。将流动注射化学发光用于药物分析不仅具有理论意义,而且还有重要的实用价值。
本文共分为两个部分。
第一部分(第一章)介绍流动注射化学发光法的应用及特点;详细介绍了几种常见的化学发光体系在药物分析中的研究应用;阐述了本章的研究内容和特点。
第二部分是研究报告(第二～六章)。
论文的第二章,研究了在碱性条件下,醋酸泼尼松能降低NBS-钙黄绿素的化学发光,其强度与醋酸泼尼松的质量浓度在一定范围内成线性关系。建立了测定醋酸泼尼松的流动注射化学发光新方法,并详细研究了影响化学发光强度的因素。在优化实验条件下,醋酸泼尼松的线性范围为2.0×10-8～1.0×10-5 g/mL,检出限(3σ)为1.3×10-8 g/mL。本法成功应用于片剂中醋酸泼尼松的测定,并对该体系的机理进行探讨(已被Analytical Letters录用)。
论文的第三章研究了在碱性条件下,雌二醇对N-溴代丁二酰亚胺(NBS)-钙黄绿素化学发光体系的化学发光具有强烈的抑制作用,其强度与雌二醇的质量浓度在一定范围内成线性关系。基于此首次建立了雌二醇的化学发光抑制分析法,并详细研究了影响化学发光强度的因素,对抑制化学发光可能的机理进行了探讨。方法对雌二醇响应的线性范围为2.0×10-8～8.0×10-6 g/mL,检出限(3σ)为8.5×10-9 g/mL,对1.0×10-7 g/mL的雌二醇进行了11次平行测定,相对标准偏差为1.2 %。方法应用于注射液中雌二醇含量的测定,结果令人满意(已在分析测试学报发表)。
论文的第四章基于在硫酸介质中,苯甲酸雌二醇可以增强KMnO4-邻菲咯啉钌的化学发光反应,进一步研究发现,Na2SO3的加入,大大增强KMnO4-邻菲咯啉钌-苯甲酸雌二醇体系的发光强度。建立了一种灵敏、快速、选择性好的测定苯甲酸雌二醇的流动注射化学发光新方法。在优化的条件下,该法线性范围ΔI=2914+11.71×108c (r=0.9961),检出限为2.4×10-8 g/mL。本法成功应用于苯甲酸雌二醇针剂的测定,并对该体系机理进行探讨(已被Luminescence录用)。
论文的第五章研究了在酸性条件下,高锰酸钾能够氧化黄体酮从而产生微弱的化学发光信号,当该反应体系中存在亚硫酸钠时,体系发光信号显著增强。建立了一种灵敏、简单、快速测定黄体酮的化学发光新方法。在优化的条件下,黄体酮的线性范围为8.0×10-8～1.0×10-6 g/mL,检出限(3σ)为4.6×10-8 g/mL。该方法已成功应用于合成样品中黄体酮含量的测定。
论文的第六章,在H2SO4介质中,酸性高锰酸钾溶液可以氧化雌二醇从而产生微弱的化学发光信号,当该反应体系中存在罗丹明B时,体系发光信号显著增强,据此建立了一种简单、快速测定雌二醇的流动注射化学发光新方法。基于这一现象,本文建立了一种化学发光测定雌二醇的新方法。该方法的线性范围为2.0×10-8～1.0×10-5 g/mL,检出限(3σ)为1.2×10-8 g/mL (3σ)。该方法已成功应用于注射液中雌二醇含量的测定。
Hormones are component of organic matters which are effective for human or animals. Hormone drugs include: glucocorticoid, adrenocorticotropic, norepinephrine, progesterone, estrogen, androgen, et al. Their common usage are intravenous, oral, topical and others. Common dosage forms include injection of powder, water injection, capsules, tablets, creams, pastes, aerosols. Hormones can be used to prevent, treat, diagnose disease and help the body return to normal function. For example, glucocorticoids mainly affects sugar and protein metabolism, and can fight inflammation.
Due to its high sensitivity, simplicity and relatively low cost, chemiluminescence analysis has been widely used in biological engineering, molecular biologic, medicine, environmental chemistry.
The thesis consists of two parts.
The first one is a brief review dealing with the research and development of chemiluminescence. The other is research reports.
In Chapter two, a nivel flow injection chemiluminescence method for the determination of prednisone acetate was developed. This method is based on that the strong CL of N-bromosuccinimide (NBS) and calcein can be inhibited by prednisone acetate. The CL intensity correlated linearly with the concentration of prednisone acetate over the range of 2.0×10-8~1.0×10-5 g/mL, the detection limit (3σ) of prednisone acetate was 1.3×10-8 g/mL. This method was applied to the determination of prednisone acetate in the tablets with good results. The mechanisms of CL reaction were also studied.
In Chapter three, a nivel flow injection chemiluminescence method for the determination of estradiol was developed. This method is based on that the strong CL of N-bromosuccinimide (NBS) and calcein can be inhibited by estradiol. The CL intensity correlated linearly with the concentration of estradiol over the ranges of 2.0×10-8~8.0×10-6 g/mL. The detection limit (3σ) of estradiol was 8.5×10-9 g/mL. This method was applied to the determination of estradiol in the injections. The mechanisms of CL reaction were also studied.
In Chapter four, a novel chemiluminescence (CL) method for the fast, simple and sensitive determination of estrogen benzoate using the reaction of tris (1,10-phenanthroline) ruthenium (II)-Na2SO3-permanganate is described. This method is based on the CL reaction of estrogen benzoate (EB) with acidic potassium permanganate and tris (1,10-phenanthroline) ruthenium (II). The CL intensity is greatly enhanced when Na2SO3 is added. After optimization of the different experimental parameters, a calibration graph for estrogen benzoate is linear in the range of 5.0×10-8~1.0×10-5 g/mL. The limits (3σ) of detection is 2.4×10-8 g/mL and the relative standard deviation was 1.3 % for 1.0×10-6 g/mL estrogen benzoate (n = 11). This proposed method was successfully applied to commercial injection samples. The mechanism of CL reaction was also studied.
In Chapter five, a new chemiluminescence method is proposed for the determination of progesterone, which is based on the reaction of KMnO4- NaSO3 progesterone system. Under the optimum conditions, the linear range of progesterone is 8.0×10-8~1.0×10-6 g/mL. The detection limit (3σ) is 4.6×10-8 g/mL for progesterone.
In Chapter six, to describe a novel flow injection chemiluminescence method for the determination of estradiol. The method is based on the oxidation of estradiol by acidic permanganate solution to give weak CL signal, which is dramatically enhanced by rhodamine B. The CL signal is linear with estradiol concentration in the range 2.0×10-8~1.0×10-5 g/mL with a detection limit of 1.2×10-8 g/mL (3σ). The method has been applied to the determination of estradiol in pharmaceutical preparations.
引文
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