共振瑞利散射技术测定多糖大分子的新方法
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摘要
摘要:共振瑞利散射(Resonance Rayleigh Scattering,RRS)是二十世纪90年代发展起来的新分析技术,它以灵敏度高(检出限可达ng/mL级)、仪器廉价易得、操作简便以及分析快速等优点引起了人们越来越多的关注。目前,这一技术已用于蛋白质、核酸、痕量金属、非金属、纳米微粒和药物分析中。多糖物质不仅是自然界中全部生命体的组成部分,还是一种重要的信息分子,在生命科学、化学、医学等学科研究中都具有举足轻重的地位。因此建立一种快速、准确、灵敏的测定方法具有重要的现实意义。
本文主要包括以下内容:
1、中性红与透明质酸钠相互作用的共振瑞利散射光谱及其分析应用在Britton-Robinson (pH 4.3～5.2)缓冲介质中,碱性吩嗪染料中性红与透明质酸钠作用形成结合产物时将导致溶液共振瑞利散射(RRS)大大增强并产生新的RRS光谱,其最大散射峰位于328 nm,另在605 nm处有一个较弱的散射峰。透明质酸钠浓度在0.08～2.5 mg/L范围内,与RRS强度有良好的线性关系。据此,建立了一种新的测定透明质酸钠的分析方法。该法具有高灵敏度,对透明质酸钠的检出限(3σ)为25.9 ng/mL,选择性也良好。应用于滴眼液和化妆水中透明质酸钠的测定,结果满意。
二、透明质酸钠-溴化十六烷基吡啶缔合物体系的共振瑞利散射光谱研究及其分析应用
在pH 4.3的Britton-Robinson缓冲溶液中,单独的透明质酸钠与溴化十六烷基吡啶的共振瑞利散射(RRS)均较弱,但当两者反应形成离子缔合物时,RRS大大增强并产生新的RRS光谱,最大RRS峰位于335 nm,另在546 nm处有一个强度较低的RRS峰。线性范围是0.09～3.0 mg/L,检出限(3σ)为29.0 ng/mL。实验表明该方法有较高的灵敏度和较好的选择性。应用于眼药水和化妆水中透明质酸钠的测定,结果满意。
三、三氨基三苯甲烷染料共振瑞利散射法测定硫酸皮肤素
在pH 5.5～6.5的Britton-Robinson缓冲溶液中,硫酸皮肤素(DS)可以与某些三氨基三苯甲烷染料如乙基紫(EV)、龙胆紫(CV)和甲基紫(MV)相互作用形成新产物,使共振瑞利散射(RRS)增强并产生新的RRS光谱。其中以最灵敏的EV-DS体系为例,其最大散射峰位于498 nm处,另在327和650 nm处有两个较弱的散射峰。硫酸皮肤素浓度在0.02～1.6 mg/L范围内,与RRS强度有良好的线性关系,检出限(3σ)为5.1 ng/mL。据此,建立了一种新的测定硫酸皮肤素的分析方法。应用于血清和尿样中硫酸皮肤素的测定,结果令人满意。
四、健那绿与硫酸软骨素相互作用的共振瑞利散射光谱及其分析应用健那绿与硫酸软骨素在pH 9.0的Britton-Robinson缓冲溶液中作用形成结合产物时将导致溶液共振瑞利散射(RRS)大大增强并产生新的RRS光谱,其最大散射峰位于326 nm处,另在407和560 nm处有两个较弱的散射峰。硫酸软骨素浓度在0.2～5.0 mg/L范围内,与RRS强度有良好的线性关系,检出限(3σ)为25.3 ng/mL。据此,建立了一种新的测定硫酸软骨素的分析方法。可用于注射液样品和片剂样品中硫酸软骨素的测定。
五、肝素-氯化十六烷基吡啶体系的共振瑞利散射光谱研究及分析应用
在pH 5.5的Britton-Robinson缓冲溶液中,肝素与阳离子表面活性剂氯化十六烷基吡啶反应形成离子缔合物时,共振瑞利散射(RRS)大大增强并产生新的RRS光谱,最大RRS峰位于293 nm,另在542 nm处有一个强度较低的RRS峰。线性范围是0.04～1.2 mg/L,检出限(3σ)为10.6 ng/mL。据此,建立了一种灵敏度高,选择性好的测定肝素的新方法。应用于肝素钠注射液的测定,结果满意。
六、十六烷基三甲基溴化铵共振瑞利散射法测定肝素及分析应用
在pH 6.0的Britton-Robinson缓冲溶液中,肝素与十六烷基三甲基溴化铵反应形成离子缔合物,使溶液共振瑞利散射(RRS)急剧增强并产生新的RRS光谱,最大RRS峰位于310 nm,并在548 nm处有一个较弱的RRS峰。肝素浓度在0.06～2.5 mg/L范围内,与RRS强度有良好的线性关系,对肝素的检出限(3σ)为19.4 ng/mL。该方法有较好的选择性,应用于肝素钠注射液的测定,结果满意。
Resonance Rayleigh Scattering (RRS) as a new technology has attracted more and more attention for its high sensitivity (detection limit of ng·mL-1 level), cheap instrument, simple operation and rapid analysis speed. At present, RRS method has been applied to the study and the determination of nucleic acids, proteins, trace metal ions, nonmetal ions, nanoparticles and some pharmaceuticals. Polysaccharides are very important component in all lives of the world, which play significant roles in the research fields of life science, chemistry and medicine as information molecular. Therefore, it is necessary to develop a speedy, accurate, highly sensitive way of analyzing polysaccharides.
The paper can be summarized as follows:
1、Resonance Rayleigh Scattering Spectra of interaction of Sodium Hyaluronate with Neutral Red and Its Analytical Application
In the Britton-Robinson buffer medium(pH 4.3～5.2), a compound complex was formed between neutral red (NR) and sodium hyaluornate (SH), leading to a great enhancement of the intensity of resonance rayleigh scattering (RRS) and giving a new RRS spectrum. The maximum scattering peak is located at 328 nm and a relatively weaker peak is located at 605 nm. It was also found that the intensity of RRS was directly proportional to the concentration of SH in the range of 0.08～2.5 mg/L. Based on these facts, a sensitive method for the determination of SH was proposed. The detection limit (3σ) of this method was found to be 25.9 ng/mL. Good selectivity of the method was shown by the results of interference test. The proposed method has been used for the determination of total amounts of sodium hyaluronate in samples with satisfactory results.
2、Resonance Rayleigh Scattering Spectral Study of Sodium Hyaluranate -Cetylpyridinium Bromide System and Its Analytical Application
In a pH 4.3 Britton-Robinson, either sodium hyaluornate (SH) or cetylpyridinium bromide (CPB) shows very faint resonance rayleigh scattering (RRS) spectra. However, when SH and CPB were mixed together, which greatly enhanced the RRS intensity and appeared a new RRS spectrum. The maximum scattering peak is at 335 nm and a relatively weaker peak is located at 546 nm. The intensity of RRS was directly proportional to the concentration of SH in the range of 0.09～3.0 mg/L, and the detection limit(3σ) was 29.0 ng/mL. The method was sensitive and selective, and has been used for the determination of total amounts of sodium hyaluronate in samples with satisfactory results.
3、Determination of Dermatan Sulfate with Some Triaminotriphenyl-methane Dyes by Resonance Rayleigh Scattering Technique
In a buffer medium of pH 5.5～6.5, dermatan sulfate (DS) can associate with some triaminotriphenyl-methane dyes such as ethyl violet (EV), crystal violet (CV) and methyl violet (MV) to form new products, resulting in a significant enhancement of the intensity of resonance rayleigh scattering (RRS) and giving new RRS spectrums. Take the most sensitive DS-EV system as an example, the maximum scattering peak is located at 498 nm and two relatively weaker peaks are located at 327 nm and 650 nm. It was also found that the intensity of RRS was directly proportional to the concentration of DS in the range of 0.02～1.6 mg/L, and the detection limit (3σ) was 5.1 ng/mL. A new method for the determination of DS was proposed. The method has achieved satisfactory results in the determination of dermatan sulfate in blood and urine.
4、Resonance Rayleigh Scattering Spectra of interaction of Chondroitin Sulfate with Janus Green and Its Analytical Application
In a pH 9.0 buffer medium, a compound complex was formed between chondroitin sulfate (CS) and janus green (JG), leading to a great enhancement of the intensity of resonance rayleigh scattering (RRS) and giving a new RRS spectrum. The maximum peak is located at 326 nm and two relatively weaker peaks are located at 407 nm and 560 nm. It was also found that the intensity of RRS was directly proportional to the concentration of CS in the range of 0.2～5.0mg/L. Based on these facts, a sensitive method for the determination of CS was proposed. The detection limit (3σ) of this method was 25.3 ng/mL. The proposed method has been used for the determination of total amounts of chondroitin sulfate in samples with satisfactory results.
5、Resonance Rayleigh Scattering Spectral Study of Heparin-Cetyl Pyridinium Chloride Monohydrate System and Its Analytical Application
In a buffer medium of pH 5.5, an ion association complex was formed between heparin (Hep) and cetyl pyridinium chloride monohydrate (CPCM), resulting in a great enhancement of the intensity of resonance rayleigh scattering (RRS) and giving a new RRS spectrum. The maximum peak is located at 293 nm and a weaker peak is located at 542 nm. It was also found that the intensity of RRS was directly proportional to the concentration of Hep in the range of 0.04～1.2 mg/L. Based on these facts, a sensitive method for the determination of Hep was proposed. The detection limit (3σ) of this method was found to be 10.6 ng/mL. The method also has good selectivity, and can be applied to determine heparin injections with satisfactory results.
6、Determination of Heparin by Resonance Rayleigh Scattering Method with Cetyl Trimethyl Ammonium Bromide and Its Analytical Application
In pH 6.0 B-R buffer solution,heparin (Hep) can react with cetyl trimethyl ammonium bromide (CTAB) to form a new product, resulting in a significant enhancement of resonance rayleigh scattering (RRS) and the appearance of a new RRS spectrum. The maximum peak is located at 310 nm and a weaker peak is located at 548 nm. The intensities of RRS are proportional to the concentration of Hep in the range of 0.06～2.5 mg/L, and the detection limit (3σ) is 19.4 ng/mL. The method also has good selectivity, and can be applied to determine heparin injections with satisfactory results.
本文主要包括以下内容:
1、中性红与透明质酸钠相互作用的共振瑞利散射光谱及其分析应用在Britton-Robinson (pH 4.3～5.2)缓冲介质中,碱性吩嗪染料中性红与透明质酸钠作用形成结合产物时将导致溶液共振瑞利散射(RRS)大大增强并产生新的RRS光谱,其最大散射峰位于328 nm,另在605 nm处有一个较弱的散射峰。透明质酸钠浓度在0.08～2.5 mg/L范围内,与RRS强度有良好的线性关系。据此,建立了一种新的测定透明质酸钠的分析方法。该法具有高灵敏度,对透明质酸钠的检出限(3σ)为25.9 ng/mL,选择性也良好。应用于滴眼液和化妆水中透明质酸钠的测定,结果满意。
二、透明质酸钠-溴化十六烷基吡啶缔合物体系的共振瑞利散射光谱研究及其分析应用
在pH 4.3的Britton-Robinson缓冲溶液中,单独的透明质酸钠与溴化十六烷基吡啶的共振瑞利散射(RRS)均较弱,但当两者反应形成离子缔合物时,RRS大大增强并产生新的RRS光谱,最大RRS峰位于335 nm,另在546 nm处有一个强度较低的RRS峰。线性范围是0.09～3.0 mg/L,检出限(3σ)为29.0 ng/mL。实验表明该方法有较高的灵敏度和较好的选择性。应用于眼药水和化妆水中透明质酸钠的测定,结果满意。
三、三氨基三苯甲烷染料共振瑞利散射法测定硫酸皮肤素
在pH 5.5～6.5的Britton-Robinson缓冲溶液中,硫酸皮肤素(DS)可以与某些三氨基三苯甲烷染料如乙基紫(EV)、龙胆紫(CV)和甲基紫(MV)相互作用形成新产物,使共振瑞利散射(RRS)增强并产生新的RRS光谱。其中以最灵敏的EV-DS体系为例,其最大散射峰位于498 nm处,另在327和650 nm处有两个较弱的散射峰。硫酸皮肤素浓度在0.02～1.6 mg/L范围内,与RRS强度有良好的线性关系,检出限(3σ)为5.1 ng/mL。据此,建立了一种新的测定硫酸皮肤素的分析方法。应用于血清和尿样中硫酸皮肤素的测定,结果令人满意。
四、健那绿与硫酸软骨素相互作用的共振瑞利散射光谱及其分析应用健那绿与硫酸软骨素在pH 9.0的Britton-Robinson缓冲溶液中作用形成结合产物时将导致溶液共振瑞利散射(RRS)大大增强并产生新的RRS光谱,其最大散射峰位于326 nm处,另在407和560 nm处有两个较弱的散射峰。硫酸软骨素浓度在0.2～5.0 mg/L范围内,与RRS强度有良好的线性关系,检出限(3σ)为25.3 ng/mL。据此,建立了一种新的测定硫酸软骨素的分析方法。可用于注射液样品和片剂样品中硫酸软骨素的测定。
五、肝素-氯化十六烷基吡啶体系的共振瑞利散射光谱研究及分析应用
在pH 5.5的Britton-Robinson缓冲溶液中,肝素与阳离子表面活性剂氯化十六烷基吡啶反应形成离子缔合物时,共振瑞利散射(RRS)大大增强并产生新的RRS光谱,最大RRS峰位于293 nm,另在542 nm处有一个强度较低的RRS峰。线性范围是0.04～1.2 mg/L,检出限(3σ)为10.6 ng/mL。据此,建立了一种灵敏度高,选择性好的测定肝素的新方法。应用于肝素钠注射液的测定,结果满意。
六、十六烷基三甲基溴化铵共振瑞利散射法测定肝素及分析应用
在pH 6.0的Britton-Robinson缓冲溶液中,肝素与十六烷基三甲基溴化铵反应形成离子缔合物,使溶液共振瑞利散射(RRS)急剧增强并产生新的RRS光谱,最大RRS峰位于310 nm,并在548 nm处有一个较弱的RRS峰。肝素浓度在0.06～2.5 mg/L范围内,与RRS强度有良好的线性关系,对肝素的检出限(3σ)为19.4 ng/mL。该方法有较好的选择性,应用于肝素钠注射液的测定,结果满意。
Resonance Rayleigh Scattering (RRS) as a new technology has attracted more and more attention for its high sensitivity (detection limit of ng·mL-1 level), cheap instrument, simple operation and rapid analysis speed. At present, RRS method has been applied to the study and the determination of nucleic acids, proteins, trace metal ions, nonmetal ions, nanoparticles and some pharmaceuticals. Polysaccharides are very important component in all lives of the world, which play significant roles in the research fields of life science, chemistry and medicine as information molecular. Therefore, it is necessary to develop a speedy, accurate, highly sensitive way of analyzing polysaccharides.
The paper can be summarized as follows:
1、Resonance Rayleigh Scattering Spectra of interaction of Sodium Hyaluronate with Neutral Red and Its Analytical Application
In the Britton-Robinson buffer medium(pH 4.3～5.2), a compound complex was formed between neutral red (NR) and sodium hyaluornate (SH), leading to a great enhancement of the intensity of resonance rayleigh scattering (RRS) and giving a new RRS spectrum. The maximum scattering peak is located at 328 nm and a relatively weaker peak is located at 605 nm. It was also found that the intensity of RRS was directly proportional to the concentration of SH in the range of 0.08～2.5 mg/L. Based on these facts, a sensitive method for the determination of SH was proposed. The detection limit (3σ) of this method was found to be 25.9 ng/mL. Good selectivity of the method was shown by the results of interference test. The proposed method has been used for the determination of total amounts of sodium hyaluronate in samples with satisfactory results.
2、Resonance Rayleigh Scattering Spectral Study of Sodium Hyaluranate -Cetylpyridinium Bromide System and Its Analytical Application
In a pH 4.3 Britton-Robinson, either sodium hyaluornate (SH) or cetylpyridinium bromide (CPB) shows very faint resonance rayleigh scattering (RRS) spectra. However, when SH and CPB were mixed together, which greatly enhanced the RRS intensity and appeared a new RRS spectrum. The maximum scattering peak is at 335 nm and a relatively weaker peak is located at 546 nm. The intensity of RRS was directly proportional to the concentration of SH in the range of 0.09～3.0 mg/L, and the detection limit(3σ) was 29.0 ng/mL. The method was sensitive and selective, and has been used for the determination of total amounts of sodium hyaluronate in samples with satisfactory results.
3、Determination of Dermatan Sulfate with Some Triaminotriphenyl-methane Dyes by Resonance Rayleigh Scattering Technique
In a buffer medium of pH 5.5～6.5, dermatan sulfate (DS) can associate with some triaminotriphenyl-methane dyes such as ethyl violet (EV), crystal violet (CV) and methyl violet (MV) to form new products, resulting in a significant enhancement of the intensity of resonance rayleigh scattering (RRS) and giving new RRS spectrums. Take the most sensitive DS-EV system as an example, the maximum scattering peak is located at 498 nm and two relatively weaker peaks are located at 327 nm and 650 nm. It was also found that the intensity of RRS was directly proportional to the concentration of DS in the range of 0.02～1.6 mg/L, and the detection limit (3σ) was 5.1 ng/mL. A new method for the determination of DS was proposed. The method has achieved satisfactory results in the determination of dermatan sulfate in blood and urine.
4、Resonance Rayleigh Scattering Spectra of interaction of Chondroitin Sulfate with Janus Green and Its Analytical Application
In a pH 9.0 buffer medium, a compound complex was formed between chondroitin sulfate (CS) and janus green (JG), leading to a great enhancement of the intensity of resonance rayleigh scattering (RRS) and giving a new RRS spectrum. The maximum peak is located at 326 nm and two relatively weaker peaks are located at 407 nm and 560 nm. It was also found that the intensity of RRS was directly proportional to the concentration of CS in the range of 0.2～5.0mg/L. Based on these facts, a sensitive method for the determination of CS was proposed. The detection limit (3σ) of this method was 25.3 ng/mL. The proposed method has been used for the determination of total amounts of chondroitin sulfate in samples with satisfactory results.
5、Resonance Rayleigh Scattering Spectral Study of Heparin-Cetyl Pyridinium Chloride Monohydrate System and Its Analytical Application
In a buffer medium of pH 5.5, an ion association complex was formed between heparin (Hep) and cetyl pyridinium chloride monohydrate (CPCM), resulting in a great enhancement of the intensity of resonance rayleigh scattering (RRS) and giving a new RRS spectrum. The maximum peak is located at 293 nm and a weaker peak is located at 542 nm. It was also found that the intensity of RRS was directly proportional to the concentration of Hep in the range of 0.04～1.2 mg/L. Based on these facts, a sensitive method for the determination of Hep was proposed. The detection limit (3σ) of this method was found to be 10.6 ng/mL. The method also has good selectivity, and can be applied to determine heparin injections with satisfactory results.
6、Determination of Heparin by Resonance Rayleigh Scattering Method with Cetyl Trimethyl Ammonium Bromide and Its Analytical Application
In pH 6.0 B-R buffer solution,heparin (Hep) can react with cetyl trimethyl ammonium bromide (CTAB) to form a new product, resulting in a significant enhancement of resonance rayleigh scattering (RRS) and the appearance of a new RRS spectrum. The maximum peak is located at 310 nm and a weaker peak is located at 548 nm. The intensities of RRS are proportional to the concentration of Hep in the range of 0.06～2.5 mg/L, and the detection limit (3σ) is 19.4 ng/mL. The method also has good selectivity, and can be applied to determine heparin injections with satisfactory results.
引文
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