皮肤晚期糖基化终末产物的自体荧光光谱特性

在分析晚期糖基化终末产物(Advanced glycation end products,AGEs)标准品三维荧光光谱和皮肤中AGEs自体荧光光谱的基础上,确定了皮肤中AGEs的最佳激发波长为370 nm.使用自行设计的皮肤荧光光谱检测装置对正常人和糖尿病患者皮肤AGEs的荧光光谱进行了检测,结果表明:在418,450...     

晚期糖基化终末产物的特性及其研究现状分析

分析了晚期糖基化终末产物的生化特性和主要结构形式,介绍了目前检测晚期糖基化终末产物的主要方法和各自的优缺点,讨论了荧光光谱检测方法。用检测系统测试了晚期糖基化终末产物的激发光谱,同时采用370nm的单色光作为激发光源,分别对正常人和糖尿病患者的皮肤进行了荧光光谱检测,通过获得的发射荧光光谱分析可以发现两者在450nm附近的荧光存在明显的差异。结果表明该荧光光谱测量系统快速、无创、简单,可应用于对糖尿病、人体衰老、氧化应激等病情进行早期预测和诊断。     

晚期糖基化终末产物荧光测量及其预校正技术研究

分析了检测晚期糖基化终末产物的必要性,利用晚期糖基化终末产物特定的激发谱和荧光发射谱,设计了皮肤晚期糖基化终末产物荧光测量装置。采用370 nm的单色光作为激发光源,分别对30例志愿者(糖尿病患者11人和对照组19人)的皮肤前臂内侧组织进行了荧光光谱检测。发现同一测试对象在前臂内侧不同部位的测试结果存在差异,肤色对测试结果也会产生很大的影响。利用光谱预校正技术消除了测量位置和皮肤肤色对测试结果的影响,结果表明该预校正技术具有一定的校正效果,重复性较好。     

晚期糖基化终末产物荧光光谱测量系统的设计

提出了一种利用荧光光谱技术测量晚期糖基化终末产物的方法,阐述了该荧光光谱测量系统的测量原理,设计并搭建了该荧光光谱测量系统。利用该系统对365,370,375,380,385 nm波段进行了激发光谱扫描测试,得出375 nm为最佳激发波长;采用375 nm的单色光作为激发光源,分别对正常人和糖尿病患者的皮肤进行了荧光光谱检测,发现两者在450 nm附近的荧光存在明显的差异。实验结果证明该荧光光谱测量系统快速、无创、简单,可应用于对糖尿病、人体衰老、氧化应激等病情进行早期预测和诊断。     

不同波长激发光对血清荧光光谱影响的实验研究

采用日本岛津荧光光度计RF5301,研究了血清的荧光光谱与激发光波长的关系。实验结果表明：在不同波长的紫外光激励下,血清产生的荧光光谱线型及峰值波长基本相同,与激励光波长无关,但荧光峰强度随激励光波长变化而变化。血清的荧光光谱有两个较强的荧光发射区,其中第一个发射区处于300～410 nm,第二个发射区处于410～530 nm。当激发光波长小于310 nm,荧光主要集中在第一发射区,荧光峰位于330和370 nm处,并产生竞争现象。当激发光波长大于250 nm时,只出现330 nm处的荧光峰,其最佳激励光波长为300 nm;当激发光波长大于320 nm,第一发射区的荧光变弱,在第二发射区的荧光变强,荧光峰位于452 nm。此研究为血液的光谱特性研究提供了实验依据,对光诱导荧光光谱诊断技术中激发光波长的选择具有一定的参考价值。     

光谱校正技术在皮肤晚期糖基化终末产物检测中的应用

利用近紫外荧光光谱测量了人体皮肤组织晚期糖基化终末产物,并针对由被测皮肤组织吸收和散射引起的光谱畸变问题,重点研究了自体荧光光谱校正技术在荧光无创检测中的应用。采用波长为370nm的单色光照射人体手臂内侧皮肤,用光纤探头收集自体荧光光谱。在同一测试位置对荧光波段进行光谱扫描,得到皮肤组织反射率曲线。通过利用皮肤反射率曲线校正相应自体荧光光谱来获得固有荧光光谱。分别对糖尿病患者和健康对照组进行光谱检测和光谱校正,分析比较不同测试对象的校正结果。实验结果表明,光谱校正技术剔除了不同测试对象的皮肤差异对自体荧光光谱的影响,获得的固有荧光光谱能够作为区分糖尿病患者和健康对照组的依据。     

配合物Eu(C5H8NO3)2(C3H5N2)2Cl3·3H2O光致发光性能

利用OPO激光激发光谱和三维荧光光谱研究了配合物Eu(C5H8NO3)2(C3H5N2)2Cl3·3H2O固体粉末在不同激发光源下的荧光特性,测试了不同浓度配合物水溶液的荧光光谱.固体荧光结果显示该配合物具有很好的荧光性能,当激发光波长为320-400nm时,产生波长分别为400-500nm、580-620nm及690-710nm的三个荧光区;当激发光波长为700-880nm时产生峰值为450nm升频转换荧光,激发光波长为700-800nm时产生峰值分别为590nm和615nm的升频转换荧光.溶液荧光结果表明在10-4-10-2mol/L浓度范围内荧光强度与溶液浓度呈正相关.对其可能的发光机制进行了探讨.     

常用合成食品色素荧光光谱研究

分析合成食品色素的分子结构特点,根据荧光与分子结构的关系,理论上推断合成食品色素是荧光物质。应用SP-2558多功能光谱测量系统,测得胭脂红、苋菜红、柠檬黄、日落黄、亮蓝等五种最常用的合成食品色素标准溶液的三维荧光光谱。结果表明,胭脂红在波长330～430 nm的光激发下,产生较强荧光,荧光峰值波长为621 nm,最佳激发波长为376 nm;苋菜红在波长300～440 nm的光激发下,产生较强荧光,荧光峰值波长为643 nm,最佳激发波长为370 nm;柠檬黄在波长280～380 nm的光激发下,产生很强荧光,荧光峰值波长为565 nm,最佳激发波长为315 nm;日落黄在波长310～410 nm的光激发下,产生较强荧光,荧光峰值波长为592 nm,最佳激发波长为348 nm;亮蓝在波长320～390 nm的光激发下,产生较强荧光,荧光峰值波长为456 nm,最佳激发波长为350 nm。进而对这五种合成食品色素的荧光光谱进行了分析讨论。结果可为食品色素检测和食品安全提供帮助。     

不同波长激发光诱导奶牛血液荧光光谱特性

实验样品取自正常奶牛静脉血液并用枸橼酸钠抗凝。实验结果表明:激发光波长不同,激发的荧光光谱线的强度和峰值不同;实验中用220～270nm波长的光激发荧光时,在317,367nm荧光主峰强度出现竞争现象;比较多种激发光激发的荧光光谱的实验结果知:用220,230,240,290,350,480,500nm波长的激发光激发下的荧光强度较强,而用其他波长的激发光激发下的荧光强度较弱,且在317,365,367,388,348,463,467,607,638nm波长附近出现比较强的峰值;合适波长的激发光对奶牛血液会有比较强的生物学效应。     

乙醚溶液荧光光谱特性及其机理研究

研究了低浓度乙醚水溶液在紫外光激励下的荧光光谱,以及其荧光特性随激发光波长和乙醚溶液浓度改变的变化规律,并对其产生机理和谱线特性进行了分析和探讨。结果显示,乙醚溶液在306 nm附近出现明显的荧光峰,其最佳激励波长为245 nm,且在292 nm处还有次峰出现。激发光波长改变时,相应的荧光峰值位置基本不变,且荧光峰的强度随激发光波长的变化呈高斯分布。荧光次峰和主峰的强度产生竞争。随浓度增加,306 nm处的荧光强度线性增强,当增至7%后,发生浓度猝灭,强度线性减弱。研究结果将为检测有毒、麻醉物质——乙醚的浓度及纯度等提供参考。     

Formation of Pentosidine Cross-Linking in Myoglobin by Glyoxal: Detection of Fluorescent Advanced Glycation End Product

Glyoxal, a reactive α-oxoaldehyde, increases in diabetic condition and reacts with proteins to form advanced glycation end products (AGEs) following Maillard-like reaction. Considering the significance of protein modification by glyoxal-derived AGEs, we investigated the in vitro effect of glyoxal (200 μM) on the monomeric heme protein myoglobin (Mb) (100 μM) after incubation for one week at 25 °C. Glyoxal-treated Mb exhibited increased absorbance around the Soret region, decreased α-helicity and thermal stability compared to control Mb. Intrinsic fluorescence spectrum of the treated Mb showed an additional signal in the 400–500 nm region on excitation at 280 nm that was absent in control Mb. When excited at 335 nm, the glyoxal-treated sample gave a strong fluorescence indicating AGE formation. Mass spectrometric studies revealed formation of glyoxal-derived fluorescent AGE adduct pentosidine between Lys-145 and Arg-139 residues of Mb. Other than pentosidine, additional AGE adducts, namely, carboxymethyllysine at Lys-133, hydroimidazolone at Arg-31 and pyrrolidone-carboxymethyllysine at Lys-145 were also detected. Lys-145 was thus found to contain two different types of AGE adducts, indicating the heterogeneous nature of in vitro glycation reaction. AGE-induced protein modifications might be associated with complications in disease conditions.     

A portable system for noninvasive assessment of advanced glycation end-products using skin fluorescence and reflectance spectrum

An optical system has been developed for noninvasive assessment of skin advanced glycation end-products (AGEs). The system comprises mainly a high-power ultraviolet light emitting diode (LED) as an excitation source, an LED array for the reflectance measurement, a trifurcated fiber-optic probe for light transmitting and receiving, and a compact spectrometer for light detecting. Both skin fluorescence of a subject and the reflectance spectrum of the same site can be obtained in a single measurement with the system. Demonstrative measurements with the system have been conducted. Results indicate that the measured reflectance spectrum can be used to compensate for the distortion of AGEs fluorescence, which is caused by skin absorption and scattering. The system is noninvasive, portable, easy to operate, and has potential applications for clinical diagnosis of AGE-related diseases, especially diabetes mellitus.     

Diagnosis of corneal pathology by laser fluorescence spectroscopy

We have studied the difference between the fluorescence spectra of the human cornea in vivo under normal conditions and after contact lenses have been worn for different lengths of time, with excitation by emission from a nitrogen laser (337 nm). The most significant sections of the difference spectrum were identified, corresponding to peaks for endogenous fluorophores (NADH and collagen). A high correlation was found between how long the contact lenses have been worn and the fluorescence intensity ratio for wavelengths 460 nm and 410 nm.     

基于荧光分析法和APTLD相结合的多环芳烃的检测

为准确进行浓度检测,用Savitzky-Golay(SG)多项式曲面平滑法去除三维荧光光谱数据的冗余信息,分别采用平行因子法(PARAFAC)算法和交替惩罚三线性分解(APTLD)算法对光谱数据进行分解。设计多环芳烃类污染物的检测实验,分析了芴(FLU)、苊(ANA)及两者混合溶液的荧光光谱特性。FLU溶液在λ_(ex)/λ_(em)=302/322 nm处存在一个明显的荧光峰,并且存在连续侧峰。ANA溶液存在两个荧光峰,分别为λ_(ex)/λ_(em)=290/322 nm和λ_(ex)/λ_(em)=290/336 nm。在激发波长200～370 nm扫描范围和发射波长240～390 nm扫描范围内,FLU和ANA荧光光谱重叠严重。结果表明,两种算法均能分辨出FLU和ANA,并取得了很高的回收率,但APTLD算法的检测效果更好。     

Fluorescence Anisotropy of Tyrosinate Anion Using One-, Two- and Three-Photon Excitation

We examined the emission spectra and steady-state anisotropy of tyrosinate anion fluorescence with one-photon (250–310 nm), two-photon (570–620 nm) and three-photon (750–930 nm) excitation. Similar emission spectra of the neutral (pH 7.2) and anionic (pH 13) forms of N-acetyl-L-tyrosinamide (NATyrA) (pK_a 10.6) were observed for all modes of excitation, with the maxima at 302 and 352 nm, respectively. Two-photon excitation (2PE) and three-photon excitation (3PE) spectra of the anionic form were the same as that for one-photon excitation (1PE). In contrast, 2PE spectrum from the neutral form showed ~30-nm shift to shorter wavelengths relative to 1PE spectrum (λ_max 275 nm) at two-photon energy (550 nm), the latter being overlapped with 3PE spectrum, both at two-photon energy (550 nm). Two-photon cross-sections for NATyrA anion at 565–580 nm were 10 % of that for N-acetyl-L-tryptophanamide (NATrpA), and increased to 90 % at 610 nm, while for the neutral form of NATyrA decreased from 2 % of that for NATrpA at 570 nm to near zero at 585 nm. Surprisingly, the fundamental anisotropy of NATyrA anion in vitrified solution at ?60 °C was ~0.05 for 2PE at 610 nm as compared to near 0.3 for 1PE at 305 nm, and wavelength-dependence appears to be a basic feature of its anisotropy. In contrast, the 3PE anisotropy at 900 nm was about 0.5, and 3PE and 1PE anisotropy values appear to be related by the cos⁶ θ to cos² θ photoselection factor (approx. 10/6) independently of excitation wavelength. Attention is drawn to the possible effect of tyrosinate anions in proteins on their multi-photon induced fluorescence emission and excitation spectra as well as excitation anisotropy spectra.     

磺基水杨酸的荧光光谱与荧光量子产率

报道了磺基水杨酸 (SSA)的荧光光谱和荧光量子产率。在 pH <2时 ,SSA无荧光 ,随pH升高 ,SSA荧光增强 ,在 pH 5～ 10 5之间 ,SSA有稳定的强荧光 ,最大发射波长为 4 0 2nm ,激发波长为 2 12 ,2 38和2 97nm。在 pH >13的强碱性条件下 ,SSA转变为另一种荧光型体 ,最大激发波长 2 6 1nm ,最大发射波长390nm。SSA浓度较高时 ,荧光激发光谱发生变化 ,但发射光谱不变。在近中性条件下 ,SSA稀溶液的荧光强度与浓度之间存在良好的线性关系 ,线性范围为 5～ 2 5 0ng·mL- 1 ,检测下限为 5ng·mL- 1 。以硫酸奎宁为参比 ,测量了SSA在不同波长下的荧光量子产率 ,在最大激发波长 2 97nm处的荧光量子产率为 0 5 4。