Study on fluorescent property of degrading products of piperacillin and its analytical application

The fluorimetric property of the degrading products of piperacillin has been studied in detail. The studies on degrading pH, degrading time, detection alkalinity and other corresponding analytical parameters of acid degradation have been made. Then fluorometry of piperacillin was established by producing its stable fluorescent products. The detection limit for acid degradation analytical method is 2.34 ng/ml, the linear range is 7.80-4.0 x 10(2) ppb. The analytical sensitivity, precision and stability of degrading products of acid degradation are satisfactory, which has been used for the determination of the trace piperacillin in human serum and urine with satisfactory results.     

Laser‐Induced Fluorometry for Capillary Electrophoresis

Laser‐induced fluorometry (LIF) has achieved the detection of single molecules, which ranks it among the most sensitive of detection techniques, whereas capillary electrophoresis (CE) is known as a powerful separation method with resolution that is beyond the reach of many other types of chromatography. Therefore, a coupling of LIF with CE has established an unrivaled analytical technique in terms of sensitivity and resolution. CE‐LIF has demonstrated excellent performance in bioanalytical chemistry for the high‐resolution separation and highly sensitive detection of DNAs, proteins, and small bioactive molecules. This review describes the CE‐LIF methods developed by the author's group that include indirect and direct detection using diode lasers, post‐column derivatization, and Hadamard transformation, as well as applications to the binding assays of specific DNA immunoassays of proteins and to the determination of anticancer drugs.     

双峰倍增配平法用于同步、导数-同步荧光法同时测定邻羟基苯甲酸和间羟基苯甲酸

邻羟基苯甲酸（ｏＨＢＡ）和间羟基苯甲酸（ｍＨＢＡ）荧光光谱严重重叠，同步及导数技术虽使选择性有所改善，但仍不能完全分辨开重叠谱。用双峰倍增配平计算法结合同步、一阶导数－同步荧光法对双组分体系（邻羟基苯甲酸／间羟基苯甲酸，ｐＨ１２介质）同时测定。结合计算的两种测定方法精密度、回收率和不同组分间浓度比范围均优于一阶导数－同步荧光法。     

Microwave-assisted on-line derivatization for sensitive flow injection fluorometric determination of formaldehyde in some foods

A rapid and sensitive flow injection fluorometry has been developed for the determination of formaldehyde based on the microwave on-line accelerating its Hantzsch reaction with cyclohexane-1,3-dione. Under the optimized conditions, the fluorescent intensity is proportional to formaldehyde content in the range from 0.05 ng/mL to 2.000 μg/mL. The detection limit (S/N = 3) is 0.02 ng/mL and the analytical frequency is 28 injections per hour. The relative standard deviations are 2.2% and 3.1% for eleven injections of 0.100 and 0.001 μg/mL of formaldehyde, respectively. With the assistance of microwave irradiation, a best sensitive fluorometry was established for the determination of formaldehyde at a high analytical frequency. This method was successfully applied to food analysis without requiring any sample pretreatment, and the determination results were correlated well with those obtained by the standard method with a sample pretreatment of steam distillation.     

[MeCl4]-型配阴离子-亚甲蓝离子缔合物的溶剂萃取和荧光法测定痕量亚甲蓝

研究了离子缔合物溶剂萃取的适宜条件、有机相中荧光和共振荧光光谱特征, 讨论了离子缔合物的组成和结构, 考察了有关的分析化学性质. 建立了测定痕量亚甲蓝的高灵敏度的方法, 其检出限分别为0.2和0.6 ng/mL(荧光法)以及1.1和2.8 ng/mL(共振荧光法), 荧光法具有更高的灵敏度, 更宜于痕量亚甲蓝的测定. 将该方法用于人血清和尿样中痕量亚甲蓝的测定, 结果较好.     

野酸枣氮掺杂荧光碳量子点高灵敏检测Hg2+

以天然产物野酸枣和色氨酸为原料,通过水热法一步合成量子产率为16.9%的氮掺杂荧光碳量子点。该碳量子点具有良好的水溶性和耐光性,在高盐环境中也呈现出了较高的稳定性。应用荧光光谱、透射电子显微镜(TEM)、傅里叶变换红外光谱(FTIR)和X射线光电子能谱(XPS)对碳量子点进行了表征。此外,Hg^2+能够有效地猝灭碳量子点的荧光,猝灭机理为电子转移的动态猝灭。基于此,可将碳量子点作为荧光探针检测Hg^2+。方法对Hg^2+的检测范围为1~50 nmol/L,检出限为0.26 nmol/L,能够应用于实际水样中Hg2+含量的测定。     

Comparative Study of Fluorometric and Electrochemical Detection of Catecholamine,Serotonin, and Metabolites in Rat Brain by High-Performance Liquid Chromatography

Abstract

A high-performance liquid chromatograph connected to a newly designed fluorometer was used for the simultaneous determination of norepinephrine, dopamine, serotonin, 3-methoxy-4-hydroxyphenylglycol (MHPG), homovanillic acid, and 5-hydroxyindoleacetic acid by measuring native fluorescence. These amines and metabolites extracted from the frontal cortex of a rat were purified by adsorption on alumina and an extraction with ethyl acetate and separated in the chromatograph with an ODS column. The detection limits ranged from 0.5 to 3 pmoles. Compared to the electrochemical (EC) detection at an electrode potential of 0.8 V, the sensitivity was almost the same or somewhat lower, but MHPG was highly fluorescent. In the fluorometry, large front peaks observed in the EC detection did not appear, and interfering peaks were not observed.     

Determination of the Protein Labeled by Dyes Based on Direct Detection of Light Intensity Absorbed

A novel spectrometric technique of molecules based on direct detection of light intensity absorbed had been proposed. This paper describes the application of this technique for the determination of the protein labeled by dyes. The result showed that the sensitivity and detection limit of this method were better than that of UV-Vis determination for the same system.     

Development of novel reagent for Hantzsch reaction for the determination of formaldehyde by spectrophotometry and fluorometry.

A novel reagent, acetoacetanilide (AAA), was introduced to the determination of formaldehyde based on Hantzsch reaction. A simple and highly sensitive fluorometric method was achieved by using AAA. The main advantages in the use of this reagent are: the reaction is carried out at room temperature without any heating system, the cyclization product based on Hantzsch reaction is soluble in water, and the product can be detected by spectrophotometry and fluorometry. The maximum absorption wavelength of the product occurs at 368 nm, and the maximum excitation and emission wavelengths are found at 370 and 470 nm, respectively. Several important experimental variables of the procedures were examined; particularly, the reaction temperature, reaction time, concentrations of reagents, and pH of the reagent solution were optimized for improving the detecting sensitivity. The calibration graph was linear in the range of 1 x 10(-7) - 1 x 10(-6) M or much higher concentrations. The limit of detection (LOD), based on three times of the standard deviation of the reagent blank, was 2.0 x 10(-8) M. The proposed method was applied to the determination of formaldehyde in environmental water samples. Many foreign species commonly existing in water samples did not interfere with the determination of formaldehyde in the proposed method.     

Highly sensitive and direct detection of DNA fragments using a laser-induced capillary vibration effect.

A pulsed laser-induced stationary wave capillary vibration detection method was applied to the sensitive detection of capillary gel electrophoresis, and the direct detection of non-labeled nucleic acids, such as DNA sequencing products, was demonstrated. An excimer laser operating at 248 nm was used as a CVL excitation source, and polynucleotides were sensitively detected without derivatization. From an investigation on the endurance of several matrixes to pulsed laser irradiation, a polyacrylamide without a cross-linker (0%C) was found to have adequate endurance, and it exhibited no serious damage during an analysis. A cytosine-terminated sequence reaction product was detected with a sensitivity close to that of laser-induced fluorometry (LIF). These results suggest the feasibility of the highly sensitive detection of ultramicro amounts of biological materials without a pre- or post-column derivatization, which has usually been required in sensitive detection procedures, such as LIF. Furthermore, the feasibility of a novel DNA sequencing method is also suggested.     

Dual-light source excitation for mode-filtered light detection

The sensitivity of a multi-channel mode-filtered light detection system has been enhanced by using a dual-light source irradiation technique. The detection system was constructed from an annular column consisting of a bare optical fibre inserted into a capillary. Sample was introduced through the gap between the fibre and the capillary. A multi-channel charge-coupled device was set on the side of the capillary at which four detection windows could be simultaneously monitored. The changes in the intensities of the mode-filtered light on exposure to various concentrations of ethanol samples from each detection window were monitored. The theoretical studies on the sensitivity of detection of the detection system using dual-light source irradiation have been described. The sensitivity of detection was enhanced when a dual-light source instead of a single-light source was employed. The working concentration range for ethanol was 0-80% (v/v) ethanol. The limit of detection was determined to be 1% (v/v) ethanol. The proposed method has been successfully applied to the determination of ethanol contents of some wine samples. The results were satisfactory compared with values obtained from a standard reference method.     

Simultaneous determination of dopamine and carvedilol in human serum and urine by first-order derivative fluorometry.

A sensitive and selective method for simultaneous determination of carvedilol and dopamine was described. The emission wavelengths of carvedilol and dopamine were at 354 nm and 314 nm with the excitation at 290 nm, respectively. The determination of carvedilol and dopamine by normal fluorometry was difficult because the emission spectra of carvedilol and dopamine were overlapped seriously. The first derivative peaks of carvedilol and dopamine were at 336 nm and 302 nm, respectively. The linear regression equations of the calibration graphs of carvedilol and dopamine were C = 0.000557H-0.00569 and C = 0.00438H-0.0812, with the correlation coefficients were 0.9953 and 0.9988, respectively. The liner range for the determination of carvedilol was 0.002 microg ml(-1) to 0.02 microg ml(-1), and 0.05 microg ml(-1) to 0.6 microg ml(-1) for dopamine. The detection limits were 1 ng ml(-1) for carvedilol and 0.04 microg ml(-1) for dopamine, respectively. The relative standard derivative (RSD) of 4.38% and 4.35% was observed for carvedilol and dopamine, respectively. The recovery of carvedilol was from 95.00% to 106.7% in human serum and from 97.50% to 105.0% in urine sample. The recovery of dopamine was from 100.0% to 102.5% in human serum and from 97.50% to 105.0% in urine sample. This method is simple and can be used for determination of carvedilol and dopamine in human serum and urine sample with satisfactory results.     

多菌灵与Pd（II）体系的共振瑞利散射光谱及分析应用

在pH=10.0的Britton-Robinson（BR）缓冲溶液中,多菌灵与Pd(Ⅱ)反应形成1∶1的六元螯合物,导致共振瑞利散射（RRS）、二级散射（SOS）和倍频散射（FDS）显著增强,并产生新的共振瑞利散射光谱,其最大RRS、SOS和FDS波长分别位于309、606和310 nm。 在一定范围内,3种散射增强（ΔIRRS、ΔISOS和ΔIFDS）均与多菌灵的浓度成正比,反应具有较高的灵敏度,对于多菌灵的检出限分别为7.1×10-9 g/mL（RRS）、7.4×10-9 g/mL（SOS）和10.7×10-9 g/mL（FDS）。 据此提出了测定多菌灵的光散射新方法。 以灵敏度最高的RRS法为例,测定了西芹和市售农药中多菌灵的含量,结果与标准方法一致。 文中还对反应机理和散射增强的原因进行了讨论。     

Derivative spectrophotometric determination of praseodymium in rare earth mixtures with lomefloxacin

Praseodymium forms a Pr(LMFX)(3) complex with lomefloxacin. In this paper, the absorption spectra of the complex has been investigated by applying conventional and derivative spectrophotometric methods. It was found that lomefloxacin could form a stable complex with praseodymium in the pH 6.5-8.5 media. The absorption intensity of the complex is 4.5-fold more than PrCl(3). Using the second derivative spectra, the sensitivity is 7.4 times higher for Pr than in the normal method (zero derivative spectra). The second derivative spectrophotometry for determination of praseodymium in the presence of other rare earths has been developed. The calibration curves were linear in the range of 3.5-65 mug ml(-1) for Pr. The detection limit is 0.85 mug ml(-1). The method is satisfactory for the determination of praseodymium in mixed rare earths.     

HPLC and chemometrics-assisted UV-spectroscopy methods for the simultaneous determination of ambroxol and doxycycline in capsule

High-performance liquid chromatography (HPLC) and multivariate spectrophotometric methods are described for the simultaneous determination of ambroxol hydrochloride (AM) and doxycycline (DX) in combined pharmaceutical capsules. The chromatographic separation was achieved on reversed-phase C(18) analytical column with a mobile phase consisting of a mixture of 20mM potassium dihydrogen phosphate, pH 6-acetonitrile in ratio of (1:1, v/v) and UV detection at 245 nm. Also, the resolution has been accomplished by using numerical spectrophotometric methods as classical least squares (CLS), principal component regression (PCR) and partial least squares (PLS-1) applied to the UV spectra of the mixture and graphical spectrophotometric method as first derivative of the ratio spectra ((1)DD) method. Analytical figures of merit (FOM), such as sensitivity, selectivity, analytical sensitivity, limit of quantitation and limit of detection were determined for CLS, PLS-1 and PCR methods. The proposed methods were validated and successfully applied for the analysis of pharmaceutical formulation and laboratory-prepared mixtures containing the two component combination.     

盐酸表柔比星与核酸相互作用的共振瑞利散射光谱研究及其分析应用

用共振瑞利散射光谱研究了盐酸表柔比星(EPI)与小牛胸腺DNA(ctDNA)、鲑鱼DNA(sDNA)、鲱鱼精DNA(hsDNA)和酵母RNA(yRNA)等核酸之间的相互作用. 实验表明在pH 2.0左右的酸性介质中, 表柔比星及核酸本身的共振瑞利散射(RRS)均十分微弱, 但是当它们相互作用形成结合产物时, 将导致RRS增强并出现新RRS光谱. 不同核酸与表柔比星结合产物的RRS 光谱特征略有差异, 散射增强程度则各不相同, 其相对散射强度的顺序是ctDNA≈sDNA＞hsDNA＞yRNA . 在一定范围内核酸浓度与散射强度成正比, 据此可以建立一种新的用表柔比星测定DNA的 RRS 法, 方法具有高灵敏度, 对于不同DNA 其检出限(3s)在24.0 ng/mL 至28.0 ng/mL 之间, 用于合成样品分析, 结果满意. 文中还研究了适宜的反应条件, 影响因素和结合产物的分析化学性质. 结合吸收光谱和荧光光谱特征对表柔比星与DNA 的反应机理进行了讨论.     

Determination of lysozyme at the nanogram level in food sample using Resonance Rayleigh-scattering method with Au nanoparticles as probe

A new method for determination of lysozyme with high sensitivity based on Resonance Rayleigh scattering (RRS) by using Au nanoparticles as a probe was proposed in this experiment. The RRS spectrum, nonlinear scattering (second-order scattering (SOS), frequency doubling scattering (FDS)) spectra and absorption spectrum of the Au nanoparticles-lysozyme system have been analyzed. In addition, the effects of several factors on scattering intensities were investigated, including pH value of solution, amount of Au nanoparticles, mixing sequence of each reagent and the coexisting substances. The results showed that the coexisting substances have little influence on the RRS intensities of the systems. Moreover, the possible mechanism for the RRS enhancement of Au nanoparticles-lysozyme system was preliminary discussed. The RRS method for determination of lysozyme has good sensitivity and selectivity with the detection limits 30.1 ng/ml. The contents of lysozyme were determined with recoveries of 98.2-105.4% and relativity relative standard deviation (RSD) of 0.3-3.7%, respectively. It proved that the method established in our study is suitable for the determination of lysozyme in synthetic sample and natural chicken egg white sample.     

曙红Y共振光散射探针测定盐酸丙米嗪

在弱酸性介质中,盐酸丙米嗪与曙红Y依靠静电引力和疏水作用力形成离子缔合物,使曙红Y溶液的吸收光谱、荧光光谱和共振光散射光谱发生变化。其中以共振光散射法灵敏度最高。据此,建立了使用曙红Y作为共振光散射探针测定盐酸丙米嗪的新方法。研究了体系的吸收光谱、荧光光谱和共振光散射光谱特征。在最大散射峰364 nm处,测得盐酸丙米嗪的线性范围为0.025～2.5μg/mL,检测限为5.32 ng/mL,并将方法用于药物中盐酸丙米嗪含量的测定。     

蛋白质测定的流动注射荧光法

根据荧光素的单体－二聚体平衡转化的原理，提出了一种检测蛋白质的方法；荧光素在表面活性剂作用下形成二聚体后荧光强度明显降低，加入蛋白质后二聚体解聚使荧光强度增加；结合流动注射法测定牛血清蛋白（BSA），该法简便，灵敏度高，对血清中蛋白质进行测定，结果令人满意。     

Determination of artesunate using reversed‐phase HPLC at increased temperature and ELSD detection

Artesunate (ART) determination can be performed by evaporative light scattering detection with mobile phase composed of CH₃CN/HCOOH 0.01 M (40:60 v/v; pH 2.85). Evaporative light scattering detection instead of UV detection allowed to improve the sensitivity and the LOD. However, the evaporative light scattering detection response of dihydro‐artemisinin appears weaker than for ART, whereas with UV detection the response of ART and dihydroartemisinin seemed similar. Constant analysis time was obtained on using the mobile phase with a flow rate of 0.5 mL/min and column temperature at 60°C instead of 0.7 mL/min at room temperature. This led to less solvent consumption. Moreover, decrease in the flow rate and increase in the column temperature were advantageous for higher sensitivity with both evaporative light scattering detection and UV detection. ART determination in rectal gel and suppositories were compared with these different detection modes and similar results were obtained.