功能化磁性粒子—高效液相色譜分離分析體液中的單胺類神經(jīng)遞質(zhì)
發(fā)布時間：2018-09-11 19:45
【摘要】：單胺類神經(jīng)遞質(zhì)包括兒茶酚胺類的去甲腎上腺素(NE)、腎上腺素(E)和多巴胺(DA),以及吲哚胺類的5-羥色胺(5-HT)等,這些神經(jīng)遞質(zhì)在生物體內(nèi)起著重要作用,其含量與人體健康密切相關(guān)。由于體液組成復雜,且單胺類神經(jīng)遞質(zhì)在體液中的含量低,因此,利用前處理過程對樣品進行除雜與富集是非常必要的。與傳統(tǒng)的樣品前處理方法相比,功能化磁性粒子由于具有易于分離、安全無毒等優(yōu)點,在醫(yī)藥、生物、環(huán)保等方面已得到廣泛應用,但是將磁性粒子用于體液中單胺類神經(jīng)遞質(zhì)分離分析方面的研究還很少。 本文成功地制備了三種不同功能化的磁性粒子,并利用多種手段對合成的功能化磁性粒子進行了表征,分析了它們的性質(zhì)。分別利用這三種功能化磁性粒子對尿液中單胺類神經(jīng)遞質(zhì)進行了分離分析研究,取得了令人滿意的效果。 論文主要分為以下四個部分： 1.對單胺類神經(jīng)遞質(zhì)、單胺類神經(jīng)遞質(zhì)的檢測方法、單胺類神經(jīng)遞質(zhì)的前處理方法進行了綜述,重點介紹了固相萃取在單胺類神經(jīng)遞質(zhì)前處理方面的應用及功能化磁性粒子的制備。 2.采用多步法制備得到苯硼酸基功能化磁性納米粒子(Fe3O4@APBA NPs),并通過透射電鏡(TEM)、傅里葉紅外光譜儀(FT-IR)、X射線衍射儀(XRD)和振動樣品磁強計(VSM)等不同的表征手段對合成的Fe3O4@APBA NPs粒子進行表征。利用Fe3O4@APBA NPs對四種單胺類神經(jīng)遞質(zhì)NE、E、DA及5-HT進行分離提取,結(jié)合高效液相色譜-電化學(HPLC-ECD)分析檢測。在1mL料液中,確定了單胺類神經(jīng)遞質(zhì)濃度為5μg/mL時,磁性粒子用量為10mg,萃取時間為5min,洗脫劑為0.02mol/L鹽酸-甲醇的最佳萃取-洗脫條件。實驗表明,Fe3O4@APBA NPs磁性粒子對三種兒茶酚胺類神經(jīng)遞質(zhì)NE、E及DA均展示了較好的提取效果,而對吲哚胺類神經(jīng)遞質(zhì)5-HT幾乎沒有提取,說明磁性粒子上的硼酸基對具有鄰位酚羥基結(jié)構(gòu)的化合物具有明顯的選擇性。在優(yōu)化條件下,將該磁性粒子用于分離富集尿樣中的NE、E和DA,結(jié)果顯示,NE、E及DA在0.01-2.0μg/mL濃度范圍內(nèi)色譜峰高與濃度呈現(xiàn)良好的線性關(guān)系,檢出限分別達到了7.8ng/mL、2.0ng/mL、7.9ng/mL,相對標準偏差均小于5.3%。將建立的方法用于人體尿樣中單胺類神經(jīng)遞質(zhì)的測定,獲得了令人滿意的結(jié)果,說明建立的Fe3O4@APBA NPs磁性粒子分離富集-高效液相色譜測定單胺類神經(jīng)遞質(zhì)的方法是一種簡單、快速、靈敏的新方法。 3.合成了二氧化鈦包裹的介孔功能化磁性納米粒子(Fe3O4@nSiO2@mTiO2NPs),并利用TEM、FT-IR、XRD、VSM和BET等表征手段對合成的粒子進行了表征分析。利用Fe3O4@nSiO2@mTiO2NPs對四種單胺類神經(jīng)遞質(zhì)NE、E、DA及5-HT進行了分離提取,對料液相pH、提取時間、緩沖溶液種類與濃度、磁性粒子用量、洗脫溶劑種類與濃度等條件進行了優(yōu)化。結(jié)果表明,合成的磁性粒子具有較高的選擇性,對NE、E及DA展示了較好的萃取效果,而對5-HT幾乎沒有萃取。在優(yōu)化條件下,該磁性粒子對NE、E和DA的富集因子范圍在8-10之間,結(jié)合HPLC-ECD分析檢測,在0.01-1.5μg/mL濃度范圍內(nèi),NE、E和DA的峰高與濃度之間呈現(xiàn)良好的線性關(guān)系,檢出限分別為11.8ng/mL、0.66ng/mL、6.0ng/mL。將建立的方法用于人體尿樣中單胺類神經(jīng)遞質(zhì)的測定,獲得了令人滿意的結(jié)果,說明建立的Fe3O4@nSiO2@mTiO2NPs磁性粒子分離富集-高效液相色譜測定單胺類神經(jīng)遞質(zhì)的方法是一種簡單、快速、靈敏的分析方法。 4.采用一步包埋法制備了羧甲基纖維素包裹的功能化磁性納米粒子(Fe3O4@CMC NPs),并利用TEM、FT-IR、XRD、和VSM等手段對合成的磁性粒子進行了表征。利用Fe3O4@CMC NPs磁性粒子對四種單胺類神經(jīng)遞質(zhì)NE、E、DA、5-HT進行分離提取,對料液相pH、提取時間、洗脫溶劑種類與濃度等條件進行了優(yōu)化。在優(yōu)化條件下,將該磁性粒子用于分離富集尿樣中的單胺類神經(jīng)遞質(zhì),結(jié)合高效液相色譜-電化學方法(HPLC-ECD)分析檢測,在線性范圍為0.01-1.5μg/mL時,建立的方法對E和DA的檢出限分別達到7.5和9.6ng/mL,說明建立的Fe3O4@CMC NPs磁性粒子分離富集-高效液相色譜法可以對尿樣中的腎上腺素和多巴胺進行簡單、快速、靈敏的分析。
[Abstract]:Monoamine neurotransmitters include catecholamine norepinephrine (NE), epinephrine (E), dopamine (DA), and indoleamine 5-hydroxytryptamine (5-HT). These neurotransmitters play an important role in organisms and their contents are closely related to human health. Because of the complex composition of humor and the presence of monoamine neurotransmitters in body fluid Compared with traditional sample pretreatment methods, functionalized magnetic particles have been widely used in medicine, biology, environmental protection and other fields because of their advantages of easy separation, safety and non-toxicity. However, magnetic particles are used in monoamine nerves in body fluids. There are few studies on the separation and analysis of transmitters.
In this paper, three kinds of magnetic particles with different functions were successfully prepared, and the synthesized magnetic particles were characterized by various means, and their properties were analyzed.
The thesis is mainly divided into four parts:
1. The methods for the detection of monoamine neurotransmitters, monoamine neurotransmitters and the pretreatment methods of monoamine neurotransmitters were reviewed. The application of solid phase extraction in the pretreatment of monoamine neurotransmitters and the preparation of functional magnetic particles were mainly introduced.
2. Phenylboronic acid-based functionalized magnetic nanoparticles (Fe3O4@APBA NPs) were prepared by multi-step method, and characterized by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and vibrating sample magnetometer (VSM). Neurotransmitters NE, E, DA and 5-HT were separated and extracted. High performance liquid chromatography-electrochemical analysis (HPLC-ECD) was used to determine the optimum conditions for the Extraction-elution of monoamine neurotransmitters in 1 mL feed solution. When the concentration of monoamine neurotransmitters was 5 ug/mL, the dosage of magnetic particles was 10 mg, the extraction time was 5 min, and the eluent was 0.02 mol/L hydrochloric acid-methanol. APBA NPs magnetic particles showed good extraction effect on three catecholamine neurotransmitters NE, E and DA, but almost no extraction of indoleamine neurotransmitter 5-HT, indicating that the boric acid group on magnetic particles had obvious selectivity for compounds with o-phenolic hydroxyl structure. The results showed that the peak heights of NE, E and DA showed a good linear relationship with the concentration in the range of 0.01-2.0 ug/mL. The detection limits were 7.8 ng/mL, 2.0 ng/mL and 7.9 ng/mL, respectively. The relative standard deviations were all less than 5.3%. The method was applied to the determination of monoamine neurotransmitters in human urine and the results were satisfactory. Satisfactory results showed that the method of separation and preconcentration of Fe3O4@APBA NPs magnetic particles and determination of monoamine neurotransmitters by high performance liquid chromatography was simple, rapid and sensitive.
3. Titanium dioxide-coated mesoporous functional magnetic nanoparticles (Fe3O4@nSiO2@mTiO2NPs) were synthesized and characterized by TEM, FT-IR, XRD, VSM and BET. Four monoamine neurotransmitters NE, E, DA and 5-HT were separated and extracted by Fe3O4@nSiO2@mTiO2NPs. The pH, extraction time, and extraction time were slowed down. The results showed that the magnetic particles had high selectivity and showed good extraction effect for NE, E and DA, but almost no extraction for 5-HT. Under the optimum conditions, the enrichment factor of the magnetic particles for NE, E and DA was in the range of 8-DA. The detection limits were 11.8 ng/mL, 0.66 ng/mL and 6.0 ng/mL respectively. The established method was applied to the determination of monoamine neurotransmitters in human urine with satisfactory results. The results showed that the peak heights of NE, E and DA showed a good linear relationship with the concentrations in the range of 0.01-1.5 ug/mL, and the detection limits were 11.8 ng/mL, 0.66 ng/mL and 6.0 ng/mL, respectively. Separation and preconcentration of monoamine neurotransmitters with nSiO2@mTiO2NPs magnetic particles by high performance liquid chromatography is a simple, rapid and sensitive method.
4. Carboxymethylcellulose-coated functionalized magnetic nanoparticles (Fe3O4@CMC NPs) were prepared by one-step embedding method and characterized by TEM, FT-IR, XRD and VSM. Fe3O4@CMC NPs magnetic particles were used to separate and extract four monoamine neurotransmitters NE, E, DA, 5-HT, pH and extraction time. Under the optimized conditions, the magnetic particles were used to separate and enrich monoamine neurotransmitters in urine samples. The detection limits of E and DA were 7.5 and 9.6 n respectively in the linear range of 0.01-1.5 ug/ml by high performance liquid chromatography-electrochemical analysis (HPLC-ECD). G/mL, indicating that the established Fe3O4@CMC NPs magnetic particle separation and enrichment-high performance liquid chromatography can be used for the analysis of adrenaline and dopamine in urine samples simple, rapid and sensitive.
【學位授予單位】：湖南師范大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R917;O657.7
本文編號：2237687
[Abstract]:Monoamine neurotransmitters include catecholamine norepinephrine (NE), epinephrine (E), dopamine (DA), and indoleamine 5-hydroxytryptamine (5-HT). These neurotransmitters play an important role in organisms and their contents are closely related to human health. Because of the complex composition of humor and the presence of monoamine neurotransmitters in body fluid Compared with traditional sample pretreatment methods, functionalized magnetic particles have been widely used in medicine, biology, environmental protection and other fields because of their advantages of easy separation, safety and non-toxicity. However, magnetic particles are used in monoamine nerves in body fluids. There are few studies on the separation and analysis of transmitters.
In this paper, three kinds of magnetic particles with different functions were successfully prepared, and the synthesized magnetic particles were characterized by various means, and their properties were analyzed.
The thesis is mainly divided into four parts:
1. The methods for the detection of monoamine neurotransmitters, monoamine neurotransmitters and the pretreatment methods of monoamine neurotransmitters were reviewed. The application of solid phase extraction in the pretreatment of monoamine neurotransmitters and the preparation of functional magnetic particles were mainly introduced.
2. Phenylboronic acid-based functionalized magnetic nanoparticles (Fe3O4@APBA NPs) were prepared by multi-step method, and characterized by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) and vibrating sample magnetometer (VSM). Neurotransmitters NE, E, DA and 5-HT were separated and extracted. High performance liquid chromatography-electrochemical analysis (HPLC-ECD) was used to determine the optimum conditions for the Extraction-elution of monoamine neurotransmitters in 1 mL feed solution. When the concentration of monoamine neurotransmitters was 5 ug/mL, the dosage of magnetic particles was 10 mg, the extraction time was 5 min, and the eluent was 0.02 mol/L hydrochloric acid-methanol. APBA NPs magnetic particles showed good extraction effect on three catecholamine neurotransmitters NE, E and DA, but almost no extraction of indoleamine neurotransmitter 5-HT, indicating that the boric acid group on magnetic particles had obvious selectivity for compounds with o-phenolic hydroxyl structure. The results showed that the peak heights of NE, E and DA showed a good linear relationship with the concentration in the range of 0.01-2.0 ug/mL. The detection limits were 7.8 ng/mL, 2.0 ng/mL and 7.9 ng/mL, respectively. The relative standard deviations were all less than 5.3%. The method was applied to the determination of monoamine neurotransmitters in human urine and the results were satisfactory. Satisfactory results showed that the method of separation and preconcentration of Fe3O4@APBA NPs magnetic particles and determination of monoamine neurotransmitters by high performance liquid chromatography was simple, rapid and sensitive.
3. Titanium dioxide-coated mesoporous functional magnetic nanoparticles (Fe3O4@nSiO2@mTiO2NPs) were synthesized and characterized by TEM, FT-IR, XRD, VSM and BET. Four monoamine neurotransmitters NE, E, DA and 5-HT were separated and extracted by Fe3O4@nSiO2@mTiO2NPs. The pH, extraction time, and extraction time were slowed down. The results showed that the magnetic particles had high selectivity and showed good extraction effect for NE, E and DA, but almost no extraction for 5-HT. Under the optimum conditions, the enrichment factor of the magnetic particles for NE, E and DA was in the range of 8-DA. The detection limits were 11.8 ng/mL, 0.66 ng/mL and 6.0 ng/mL respectively. The established method was applied to the determination of monoamine neurotransmitters in human urine with satisfactory results. The results showed that the peak heights of NE, E and DA showed a good linear relationship with the concentrations in the range of 0.01-1.5 ug/mL, and the detection limits were 11.8 ng/mL, 0.66 ng/mL and 6.0 ng/mL, respectively. Separation and preconcentration of monoamine neurotransmitters with nSiO2@mTiO2NPs magnetic particles by high performance liquid chromatography is a simple, rapid and sensitive method.
4. Carboxymethylcellulose-coated functionalized magnetic nanoparticles (Fe3O4@CMC NPs) were prepared by one-step embedding method and characterized by TEM, FT-IR, XRD and VSM. Fe3O4@CMC NPs magnetic particles were used to separate and extract four monoamine neurotransmitters NE, E, DA, 5-HT, pH and extraction time. Under the optimized conditions, the magnetic particles were used to separate and enrich monoamine neurotransmitters in urine samples. The detection limits of E and DA were 7.5 and 9.6 n respectively in the linear range of 0.01-1.5 ug/ml by high performance liquid chromatography-electrochemical analysis (HPLC-ECD). G/mL, indicating that the established Fe3O4@CMC NPs magnetic particle separation and enrichment-high performance liquid chromatography can be used for the analysis of adrenaline and dopamine in urine samples simple, rapid and sensitive.
【學位授予單位】：湖南師范大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R917;O657.7
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