Capillary electrophoresis coupled with electrochemiluminescence for determination of cloperastine hydrochloride

Objective To investigate the electrochemiluminescence (ECL) behavior of cloperustine hydrochloride.Methods ECL intensity of tris (2,2'-bipyridyl) rutheniumo(Ⅱ) was enhanced, the method for the determination of dupernstine hydrochloride was established using capillary electrophoresis (CE) coupled with electrochemilumolinescence (ECL) detection. Results Under the optimum conditions, ECL intensity varied linearly with cloperastine hydrochloride concentration from 7.0×10-6 g/mL to 1.0×10-4 g/mL. The detection limit (S/N=3) was 8.05×10-7g/mL. he relative standard deviation of the ECL intensity and the migration time for 11 consecutive injections of 1.0 ×10-5g/mL cloperastine hydrochloride was 2.9% and 1.5%, respectively. This method was successfully applied to eloperastine hydrochloride tablet determination. Conclusion The method has been established, validated and applied for determination of cloperastine hydrochloride.     

CE-ECL法同时测定复方磷酸可待因中三种成分的方法学研究

目的 建立毛细管电泳-电化学发光法分离并检测复方磷酸可待因中三种成分,磷酸可待因、马来酸溴苯那敏和盐酸麻黄碱含量的新方法.方法 基于碱性介质中,复方磷酸可待因溶液中三种主要成分对联吡啶钌在铂电极上的电化学发光信号的增敏作用,且增敏强度和药物浓度成线性关系,与毛细管电泳联用,建立了毛细管电泳-电化学发光法分离并检测其含量的新方法.结果 在优化的实验条件下,确定三种药物对照品的线性范围、线性方程和检出限.通过对浓度为1.0×10~(-5)kg/L的磷酸可待因、马来酸溴苯那敏和盐酸麻黄碱对照品进行12次平行测定,其ECL发光强度的RSD值分别为2.89%,3.76%,3.32%.结论 该方法可用于复方磷酸可待因口服溶液中三种主要成分的含量测定,待测样品中的基质不干扰测定,方法的回收率分别为100.4%,101.9%,99.8%(n=5).     

DETERMINATION OF TETRACYCLINE AND OXYTETRACYCLINE BY FLOW-INJECTION CHEMILUMINESCENCE METHOD

Tetracycline and oxytetracycline are broad-spec-trumantibiotics.They are not only used in humanpathologies,but alsoin veterinary medicine,ani malnutrition and feed additives for cattle breeding.In the past few years,the deter mination meth-ods of tetracycline and oxytetracycline were repor-ted,which involved in difference spectrophotome-try[1],HPLC[2,3],spectrophotometry[4-7],HPLC-MS[8-9],spectrofluori metry[10],solid-phase extrac-tion[11]and kinetic methods.Recently,flow-injection CL met…     

Luminol-potassium permanganate chemiluminescence system for the determination of three anthracycline antibiotics

Objective To establish a flow-injection chemiluminescence method for the determination of doxorubicin, epirubicin and mitoxantrone and study its reaction mechanism. Methods In alkaline medium, chemiluminescence of luminol-potassium permanganate system could be inhibited obviously by anthracycline antibiotics. Combined with flow-injection technique, a new chemiluminescence method for determining the anthracycline antibiotics was set up. The chemiluminescence mechanism of the luminol-potassium permanganate system was also discussed. Results Under optimal conditions, the good linear ranges of doxorubicin, epirubicin and mitoxantrone were 5.0×10-9-1.0×10-6g/mL, 1.0×10-9-1.0×10-5g/mL and 3×10-9-1.0×10-6g/mL, respectively. The detection limits of doxorubicin, epirubicin and mitoxantrone were 3.0×10-9g/mL, 5.0×10-8g/mL and 2.0×10-9g/mL, respectively. During eleven repeated inter-day and intra-day precision tests of 1.0×10-6g/mL samples, the relative standard deviations corresponded to reference values of 3.0%, 2.8% and 2.1%. Conclusion The developed method is sensitive, accurate, rapid and of low cost. It can be applied to determine doxorubicin hydrochloride, epirubicin hydrochloride and mitoxantrone hydrochloride in injection preparations.     

Flow-injection-enhanced chemiluminescence method for the determination of three β-blockers

Objective To develop a rapid, simple and sensitive chemiluminescence method for the determination of three β-blockers (bisoprolol, atenolol and propranolol). Methods The chemiluminescence of cerium (Ⅳ)-sulfite system was obviously sensitized by adding anyone of three β-blockers in acid media. A new chemiluminescence method was set up by combining with flow-injection technique and used to determine the three β-blockers. Results Good linear ranges were obtained at the concentrations of 2.0×10-7g/mL-4.0×10-5g/mL, 1.0×10-7g/mL-3.0×10-5g/mL and 7.0×10-7g/mL-1.0×10-5g/mL, respectively, with the detection limits of 5.0×10-8g/mL, 7.0×10-8g/mL and 5.0×10-8g/mL (S/N=3), respectively, and the relative standard deviations for 11 times consecutive injections of 1.0×10-6g/mL bisoprolol, atenolol and propranolol were 3.57%, 2.21% and 2.26%, respectively. Conclusion The developed method is sensitive, accurate, rapid and of low cost. And it can be applied to determine bisoprolol, atenolol and propranolol in pharmaceutical preparations.     

Capillary electrophoresis with inhibited electrochemiluminescent detection for the trace analysis of epinephrine and dopamine

In this paper, a novel electrochemiluminescent (ECL) detection approach was developed for highly sensitive detection of ECL inhibitors based on the ECL inhibition of Ru(bpy)32+/2-(Dibutylamino)ethanol (DBAE) system. A microfluidic ECL detection cell was fabricated to couple with the capillary electrophoresis system, the electrochemical system and the postcolumn injection system. Both Ru(bpy)32+ and DBAE solutions were injected directly to the working electrode surface by a micro-infusion system to obtain a high and stable ECL signal. The performance of this setup was demonstrated by the analysis of two typical ECL inhibitors, dopamine and epinephrine. Under the optimal conditions, the limit of detection (LOD) for dopamine and epinephrine was 50nM and 5nM respectively. The proposed method was also successfully used for the trace analysis of dopamine and epinephrine in human serum samples.     

流动注射-化学发光法测定氢化可的松

目的确定以高锰酸钾-亚硫酸钠体系测定氢化可的松的流动注射-化学发光分析方法。方法在酸性条件下,氢化可的松对高锰酸钾-亚硫酸钠体系发光反应具有明显的增敏作用。据此,建立了流动注射化学发光测定氢化可的松的分析方法。结果在优化的实验条件下,氢化可的松质量浓度在1.0×10-9-1.0×10-6g/mL范围内与发光强度呈良好的线性关系,检出限(3R)为4.0×10-10g/mL,对氢化可的松进行11次平行测定,其相对标准偏差为2.2%。结论本方法应用于注射液中氢化可的松含量的测定,快速、准确、简便,灵敏度高、线性范围宽。     

Spectrophotometric detection of diethylstilbestrol on the basis of the plasmon resonance absorption of silver nanoparticles

In this study, a spectrophotometric detection method for diethylstilbestrol (DES) was proposed by reducing silver nitrate (AgNO3) to obtain silver nanoparticles (AgNPs) in the medium of ammonia and sodium hydroxide. It was found that the resulting AgNPs have plasmon resonance absorption (PRA) characteristic at 415 nm, and the PRA is proportional to the increase of DES concentration in the range of 4.0×10-8-1.0×10-5M with the detection limit (3σ) of 1.2×10-7M. Most of the coexisting substances at high concentrations did not affect the detection of real samples, such as tablets. The recovery was in the range of 96.01%-107.41% and the RSD was lower than 4.7%. This method can be successfully applied to control preparation quality of DES.     

流动注射化学发光法测定吩噻嗪类药物

目的确定以鲁米诺-高碘酸钾发光体系测定吩噻嗪类药物的化学发光分析方法。方法在碱性介质中,盐酸氯丙嗪和盐酸异丙嗪对鲁米诺-高碘酸钾发光体系有明显的增敏作用,且增敏效果与其浓度呈良好的线性关系。基于此,建立了盐酸氯丙嗪和盐酸异丙嗪的流动注射化学发光分析方法。结果在优化的实验条件下,盐酸氯丙嗪和盐酸异丙嗪的线性范围分别为3.0×10-9-1.0×10-6g/mL和3.0×10-8-7.0×10-5g/mL,检测限分别为5.0×10-10g/mL和7.3×10-9g/mL。结论本方法简便、快速、准确、灵敏度高、线形范围宽,应用于相应注射剂和片剂分析,并与药典方法进行对照,结果令人满意。     

RP-HPLC method for the simultaneous determination of daidzein,genistein and formonetin in Solanum Lyratum Thunb

A rapid method for the simultaneous determination of daidzein, genistein and formonetin in solanum Lyratum Thunb by high performance liquid chromatography (HPLC) was developed. Separation was achieved on a Diamonsil C18 column (250 mm×4.6 mm, 5 μm) with isocratic elution, using a mobile phase of methanol-tetrahydrofuran-water (44∶3∶53, v/v). The wavelength was set at 260 nm and column was maintained at 35 ℃. The linear ranges of daidzein, genistein and formonetin were 1.0-40.0, 0.1-4.0 and 0.1-4.0 μg/mL, respectively. The average recoveries were between 98.4% and 101.3%. This method could be used for the quality control of Solanum lyratum Thunb due to its simplification, reliability, rapidity and excellent precision.     

奋乃静药物的流动注射化学发光测定方法的建立

目的建立快速测定奋乃静的流动注射化学发光新方法。方法在硝酸介质中,奋乃静能被硫酸铈氧化生成发光物质奋乃静砜,从而产生化学发光。基于此,建立了奋乃静的流动注射化学发光分析方法。结果在优化的实验条件下,不用任何发光增敏剂,奋乃静在1.0×10-7-7.0×10-5g/mL范围内与化学发光强度呈良好的线性关系,检出限(3σ)为8.0×10-8g/mL,对1.0×10-6g/mL的奋乃静进行了11次平行测定,其相对标准偏差为1.8%。结论本方法简便、快速、准确、灵敏度高、线形范围宽,应用于奋乃静片剂分析,并与药典方法进行对照,结果满意。     

基于聚苯撑苯并二恶唑的多功能光电器件

首次报道了基于全共轭高分子材料聚苯撑苯并二恶唑（PBO）作为空穴传导材料及电子给体，富勒烯（C60）作为电子传输层及电子受体的双层异质结器件．研究发现：该结构器件具有明显的光伏响应特征，开路电压达0．76V，短路电流0．07mA／cm^2；而且在正向偏压下，器件还表现出了较强的激基复合物电致发光现象，在7．1V偏压下，器件的亮度达5．3×10^3cd／m^2，而器件的最高发光效率可达1．8cd／A．该研究表明，基于PBO／C60双层异质结的多功能光电器件具有较好的开发价值和应用前景．     

基于FBG的沥青混合料横向流动变形评价

为了分析沥青混合料横向流动变形, 进行了沥青混合料的车辙试验, 利用布设于沥青混合料板表面的光纤布拉格光栅传感器, 研究了沥青混合料表面的横向应变规律; 以最大应变和蠕变稳定阶段横向应变速率绝对值为评价指标, 分析了沥青混合料横向流动变形。分析结果表明: 横向流动变形随沥青混合料的最大应变和横向应变速率绝对值的减小而降低; 横向流动变形在循环轮载作用下不断发展, 测试点距离轮载愈近其流动变形愈剧烈; 当胶粉掺量分别为0、15%、18%时, 距离轮载63 mm的测试点横向应变速率分别为6.8×10-6、4.0×10-7、6.4×10-6 min-1, 因此, 掺15%胶粉的沥青混合料具有较大的抵抗高温横向流动变形的能力; 对于15%胶粉掺量的沥青混合料, 当其集料级配分别为AC-13粗级配和AC-13细级配时, 距离轮载28 mm的测试点横向应变速率分别为6.0×10-7、7.7×10-6 min-1, 因此, AC-13粗级配沥青混合料高温抗横向流动变形能力优于AC-13细级配; 胶粉改性沥青混合料最大应变为1.96×10-4, 而胶粉和抗车辙剂复合改性沥青混合料最大应变只有1.22×10-4, 说明在高温情况下, 胶粉和抗车辙剂复合改性沥青混合料整体结构强度较大, 能够承受来自轮载的直接作用而不向轮迹两边产生横向推移致使发生较大的横向流动变形。基于光纤布拉格光栅横向应变的沥青混合料横向流动变形评价能较好地说明不同材料和级配对沥青路面产生侧向流动变形规律的影响。      

Simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture by high performance liquid chromatography

A reversed phase high performance liquid chromatography （HPLC） method was established for the simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture. A Grace Apollo Cl8 column （250 mm × 4.6 mm, 5 μm） was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid （0.2%, v/v）. Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃. An ultraviolet （UV） detector was used with a selected wavelength of 240 nm. Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12, 13-dihydroxyeuparin （r〉0.9999） and 106.9-1068.9μg/mL for glycyrrhizic acid （r〉0.9999）, respectively. Recoveries were 102.18% for 12, 13-dihydroxyeuparin and 101.17% for glycyrrhizic acid. The method developed could be applied to the simultaneous determination of 12, 13- dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.     

Preparation of recombinant firefly luciferase by a simple and rapid expression and purification method and its application in bacterial detection

A simple and rapid expression and purification method of recombinant firefly luciferase was developed for bacteria detection. A modified luciferase gene from North American firefly Photinus pyralis was cloned into pET28a expression vector and the recombinant protein was produced in Escherichia coli BL21. The recombinant luciferase, equipped with a polyhistidine affinity tag, was purified by immobilized metal ion affinity chromatography (IMAC). The approach generated an abundant expression and an efficient purification of a recombinant luciferase with final yield 1.995mg/L of cell culture. Experiments on the recombinant luciferase also showed that the relative light units (RUL) of the enzyme were 5.8×108, and the specific activity was 2.9×1010 RLU/mg. By applying adenosine triphosphate (ATP) bioluminescence to detection of the coin bacteria using the recombinant protein, the ATP content of bacteria was 9.48×10-16mol/mL, and was identical to the bacteria counts (4500CFU/mL) in order of magnitude. Taken together, our results provided a simple and efficacious method of the preparation of recombinant luciferase, which could be applied in the determination of bacteria via ATP bioluminescence.     

二茂铁对双层类脂膜的修饰及对抗坏血酸的催化研究

采用二茂铁对双层类脂膜进行修饰,考察了修饰膜的电化学行为,发现二茂铁在双层类脂膜中起到跨膜传递电子的作用。进一步研究抗坏血酸在二茂铁-双层类脂膜上的电催化氧化反应,发现氧化峰电流与其浓度在5.0×10-6～5.5×10-3mol·L-1范围内呈良好的线性关系,检测下限为1.0×10-6mol·L-1,可用于抗坏血酸的定量检测。     

沥青混合料蠕变损伤模型与损伤演化

为定量描述沥青混合料的蠕变特性,考虑沥青混合料在整个蠕变过程中同时存在蠕变硬化机制和蠕变损伤劣化机制,基于分数阶微积分理论,发展了一种相对简单的分数阶蠕变损伤模型,用分数阶Maxwell模型来描述蠕变硬化机制,用损伤应变来表示蠕变损伤劣化机制,并从统计学角度推导出沥青混合料的损伤演化方程;对AC-13沥青混合料进行了不同应力水平(0.179、0.358、0.448、0.537和0.716 MPa)下的单轴压缩蠕变试验,通过Levenberg-Marquardt优化算法进行了非线性拟合,确定了不同应力水平下分数阶蠕变损伤模型的参数与损伤演化曲线;为构建不同应力水平下统一的损伤演化模型,提出了一种统计量化沥青混合料损伤演化的方法,建立了蠕变损伤与损伤应变之间的演化关系。研究结果表明:在不同应力水平下,提出的分数阶蠕变损伤模型与试验结果的判定系数均不小于0.995,适用于描述包括衰减蠕变阶段、稳定蠕变阶段和加速蠕变阶段的整个蠕变过程;在衰减蠕变阶段,不同应力水平下沥青混合料的损伤都小于1.0×10-3,相对于蠕变破坏时的损伤0.8可以忽略不计,而进入稳定蠕变阶段以后,损伤逐渐增大;当沥青混合料的蠕变应力超过一定值时会发生蠕变破坏,其流值时间取决于所施加的应力水平;用二参数Weibull分布函数拟合所得的蠕变损伤与损伤应变之间演化关系的判定系数为0.992,说明可以建立不同应力水平下的统一损伤演化模型,且其参数只与材料性能和温度有关,与施加应力大小无关。      

Analysis of ganciclovir and its related substances using high performance liquid chromatography and liquid chromatography-mass spectrometry methods

Objective High performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC/MS) methods were developed for the determination of ganciclovir and its related substances. Methods A Hypersil ODS2 column (4.6mm×250mm, 5μm) was used with a mobile phase of 0.02M potassium 1.0mL/min, and UV detector set at 254nm was used for monitoring the eluents. Results The method was simple, rapid, selective and capable of separating all related substances at trace level with a detection limit of 0.04μg/mL. It has been validated with respect to accuracy, precision, linearity, and limits of detection and quantification. The linearity range was 10.2-153.0μg/mL with r=0.9998. The percentage recoveries ranged from 96.7% to 101.6%, and RSD was 1.24%-1.96% (n=5). Conclusion The method was found to be suitable not only for monitoring the reactions during the process development but also for quality control of ganciclovir. For identification of related substances, LC/MS was used. The mainly related substances of ganciclovir active pharmaceutical ingredients (API) were determined as guanine, (1, 3-dioxolan-4-yl) methyl acetate, and diacetyl guanine.     

碘化二甲基木防己碱对蟾蜍腹直肌N_2受体的动力学研究

本文研究了碘化二甲基木防己碱(DTI)对N_2受体的拮抗作用,并以箭毒为对照。在DTI的作用下,ACh对蟾蜍腹直肌的累加剂量—效应％曲线平行右移,测得其pA_2为5.73,K_B为2.90X 10~(-6)M;而箭毒分别为5.89和2.84X 10~(-6)M。以log(~(X-1)对DTI浓度的负对数(-log~[B])作图,其回归线斜率为-0.92;箭毒的回归线斜率为-0.95,均同竞争性拮抗剂的理论斜率-1相接近。结果表明,DTI对N_2受体的拮抗性质为竞争性拮抗、其对N_2受体的亲和力与箭毒相当。     

RP-HPLC method for the quantitation of β-Sitosterol in Elaeagnus Gon yanthes Benth

Objective To establish an RP-HPLC method for the determination of β-Sitosterol in Elaeagnus Gonyanthes Benth. Methods The separation was performed on a luna C8 (2) (150 mm×4.6 mm, 5μm) column with the mobile phase of methanol-water (88∶12, v/v) at a flow rate of 1.0 mL/min, the detection wavelength was set at 210 nm, and the temperature of the column was maintained at 35 ℃. Results The calibration curve of β-Sitosterol was linear over the concentration range of 0.075-0.375 mg/mL (r=0.9999) and the average recovery of β-Sitosterol was 96.30% with RSD of 3.60%(n=3). Conclusion The method is simple, rapid, and accurate, and can be used for the quality control of Elaeagnus Gonyanthes Benth.