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.     

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。     

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…     

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。     

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.     

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。     

Electrogenerated chemiluminescence sensor for the determination of metoclopramide using ordered mesoporous carbon for immobilizing tris(2, 2'-bipyridyl) ruthenium

A novel electrogenerated chemiluminescence (ECL) sensor for the determination of metoclopramide was developed by employing ruthenium complex as an ECL signal producer and an ordered mesoporous carbon (OMC) material as modified material. The ECL sensor was fabricated by adsorption ruthenium complex into a mixture of OMC and Nafion, which showed good electrochemical and ECL behaviors. It was found that the ECL intensity of the sensor fabricated was greatly enhanced in the presence of metoclopramide. Based on this finding, a highly sensitive and reproducible ECL method was developed for the determination of metoclopramide. The result showed that the ECL intensity was linear with the concentration of metoclopramide in the range from 1.0×10-10 to 5.0×10-7M and the detection limit was 3×10-11M. The ECL sensor exhibited a long-term stability and a fine reproducibility with relative standard deviation of 1.0 % for 1.0×10-10M metoclopramide in 18 continuous determinations. The developed method has been applied to the determination of metoclopramide in tablet samples with satisfactory results.     

核桃壳对模拟废水中Cr(Ⅵ)的动态吸附特性研究

采用废弃核桃壳对Cr(Ⅵ)浓度为20 mg·L-1的模拟废水进行了动态吸附实验.实验结果表明,在室温条件下用粒径为1.0～1.6 mm的核桃壳吸附剂用量为5.0 g、介质pH值为1.0、水样流速为3mL· min-1吸附100mL,Cr(Ⅵ)浓度为20 mg·L-1的模拟污水时,吸附后的废水中Cr(Ⅵ)浓度为0.224 mg·L-1,符合《污水综合排放标准》GB8978-1996的标准.Cr(Ⅵ)的去除率达到98.88％.同时对模型的拟合进行了实验说明,Thomas模型能较好地反映吸附过程特征.从活性穿透曲线中可见,吸附时间为142 min达到吸附穿透点,810 min达到吸附衰竭点.     

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.     

Analysis and determination of diterpenoids in unprocessed and processed Euphorbia lathyris seeds by HPLC-ESI-MS

Euphorbia lathyris （Caper spurge） is a toxic and potent Chinese materia medica （T/PCMM）. This study sought a method for identifying five diterpenoids （Euphorbia factors LI-L3, L7a, and Ls） with the spectra of UV and mass, quantifying three diterpenoids L1, L2, and L8 in crude extracts of unprocessed and processed E. lathyris seeds by liquid chromatography/ electrospray ionization mass spectrometry （LC-ESI-MS）. The analysis was achieved on an Agilent Eclipse XDB-C18 column （4.6 mm× 150mm i.d., 5 μm） with an isocratic elution with a mobile phase consisting of water and acetonitrile at a flow rate of 0.25 mL/min at column temperature of 30 ℃ and UV detection was set at 272 nm. An ESI source was used with a positive ionization mode. The calibration curve was linear in the ranges of 9.9-79 μg/mL for Euphorbia factor Lb 3.8-30.5μg/mL for Euphorbia factor L2, and 1.0-20.6 μg/mL for Euphorbia factor LB. The average recoveries （n=6） of three diterpenoids were 98.39%, 91.10% and 96.94%, respectively, with RSD of 2.5%, 2.4% and 2.1%, respectively. The contents of the three diterpenoids in processed E. lathyris seeds were 3.435, 1.367 and 0.286 mg/g, respectively, which decreased more sharply than those in unprocessed E. lathyris seeds which were 4.915, 1.944 and 0.425 mg/g, respectively. The method is simple, accurate, reliable and reproducible, and it can be applied to control the quality of unprocessed and processed E. lathyris seeds.     

治带片中苦参碱及氧化苦参碱的HPLC法测定

目的建立治带片中苦参碱及氧化苦参碱的高效液相色谱(HPLC)测定方法。方法色谱柱:Lichrospher-NH2(4.6 mm×250 mm,5μm),C18保护柱;流动相:乙腈-无水乙醇-0.5 mol/L磷酸水溶液(80∶10∶10);检测波长212 nm;流速1.0 mL/min;柱温:室温;进样量20μL。结果苦参碱和氧化苦参碱线性范围均为1.0-10.0μg/mL,回归方程苦参碱为:C=1.201×10-4A+0.161,r=0.9992;氧化苦参碱为:C=1.366×10-4A+0.221,r=0.9996,平均回收率分别为99.9%和99.4%,RSD分别为1.48%和4.33%。结论本法简便快捷,结果准确,可用于该制剂的质量控制。     

熊果酸对大鼠子宫平滑肌收缩的影响

目的观察熊果酸对离体大鼠子宫活动的影响。方法将离体子宫肌条在1 g的前负荷下温育,加入熊果酸1.0×10-8、2.0×10-8、3.0×10-8、4.0×10-8g/L,以等容量生理盐水作为自身对照,记录30 min;分别使用不同的受体拮抗剂5 min后,再加入熊果酸3.0×10-8g/L。用Biolap 410生物机能仪记录熊果酸对离体子宫平滑肌运动的影响。结果熊果酸可增强离体子宫平滑肌的收缩振幅、频率和持续时间,熊果酸剂量与子宫肌兴奋作用存在剂量-反应关系。并使用受体阻断剂苯海拉明(2×10-6mol/L)、酚妥拉明(2×10-6mol/L)、异博定(2×10-7mol/L)及消炎痛(2×10-5mol/L)观察其改变。该增强作用可被消炎痛阻断,而异博定、酚妥拉明、苯海拉明未能阻断熊果酸的作用。结论熊果酸增强作用与前列腺素酶的合成与释放有关,不通过L型电压依赖性钙通道、α受体以及H1受体发挥作用。     

尾加压素Ⅱ促进乳鼠心肌成纤维细胞分泌胶原及增殖的细胞内信号转导机制

目的探讨尾加压素Ⅱ(UⅡ)促进乳鼠心肌成纤维细胞(CFs)分泌胶原及增殖的细胞内信号转导机制。方法体外培养CFs,采用免疫组织化学染色法及羟脯氨酸测定法分别观察不同浓度UⅡ作用下,CFs中磷酸化ERK1/2细胞灰度和CFs培养上清中胶原含量的变化;蛋白激酶C(PKC)抑制剂chelerythrine chloride(Che)、ERK1/2抑制剂PD98059和钙调神经磷酸酶(CaN)抑制剂cyclosporin A(CsA)各自对UⅡ诱导的细胞增殖的影响。结果在1×10-10、1×10-9、1×10-8mol/L UⅡ作用下,CFs培养上清中胶原含量均较对照组明显增加,而CFs中磷酸化ERK1/2细胞灰度均较对照组有显著降低(P<0.01);在1×10-7mol/L UⅡ作用下,上述各参数与对照组比较无统计学意义(P>0.05)。1×10-6mol/L Che+1×10-8mol/L UⅡ组、1×10-5mol/L PD98059+1×10-8mol/L UⅡ组和5μg/mLCsA+1×10-8mol/L UⅡ组的胶原含量均高于对照组而低于1×10-8mol/L UⅡ组(P<0.05)。1×10-6mol/L Che+1×10-8mol/L UⅡ组、1×10-5mol/L PD98059+1×10-8mol/L UⅡ组和5μg/mL CsA+1×10-8mol/L UⅡ组的p-ERK1/2的灰度低于对照组(P<0.01);1×10-6mol/L Che+1×10-8mol/L UⅡ组和5μg/mL CsA+1×10-8mol/LUⅡ组的p-ERK1/2的灰度高于1×10-8mol/L UⅡ组(P<0.01),而1×10-5mol/L PD98059+1×10-8mol/L UⅡ组与之比较则无统计学意义(P>0.05)。结论UⅡ具有促进CFs分泌胶原及增殖的作用,其作用可能是通过PKC/MAPK/CaN途径实现的。     

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。     

肝硬化大鼠原位肝移植血流动力学改变

目的 研究肝硬化大鼠异体原位肝移植术后血流动力学改变.方法 雄性健康SD大鼠50只,采用腹腔及皮下注射四氯化碳(CCl_4),同时饮用乙醇溶液诱导10周,制备肝硬化大鼠模型;按KAMADA二袖套法(门静脉和肝下下腔静脉)对肝硬化大鼠原位肝移植;观察肝移植前后门静脉血流动力学的变化.结果 制备肝硬化大鼠模型36只;进行肝移植手术26次,术后成活并进行门静脉血流动力学指标检测12只.经统计学方法检验,肝移植术后门静脉压较术前明显降低[(1.60±0.10)kPa vs. (1.70±0.25)kPa,P<0.05];门静脉血流量明显增加[(7.55±1.02)mL/min vs. (6.15±0.88)mL/min,P<0.05];门静脉阻力明显降低[(11.84×10~(-2)±2.51×10~(-2))kPa/(mL·min) vs. (16.29×10~(-2)±2.40×10~(-2))kPa/(mL·min),P<0.05].结论 异体肝移植术可改善肝硬化大鼠血流动力学指标.     

Simultaneous determination of berberine, matrine and oxymatrine in traditional Chinese medicines by using nonaqueous capillary electrophoresis

A rapid method for the simultaneous determination of berberine (BBR), matrine (MT) and oxymatrine (OMT) by nonaqueous capillary electrophoresis (NACE) was developed. Optimum separation of the analytes was obtained on a 50cm×50μm i.d. fused-silica capillary using a non-aqueous buffer system of 70mM ammonium acetate, 7.0% acetic acid and 10% acetonitrile at 25kV and 20℃. The relative standard deviations (R.S.D.) of the migration times and peak areas of the three active components were 0.06%-0.20% and 0.12%-3.41% for berberine, 0.11%-0.60% and 0.74%-1.63% for matrine, 0.15% and 0.45% for oxymatrine, respectively. Detection limits of berberine, matrine and oxymtrine were 0.18μg/mL, 4.08μg/mL and 4.16μg/mL, respectively. In the tested concentration range, good linear relationships (0.9992 for berberine, 0.9988 for matrine and 0.9988 for oxymatrine) were observed. The linear calibration ranges were 0.45-360.0μg/mL for berberine, 8.16-408.0μg/mL for matrine and 20.8-416.0μg/mL for oxymatrine. This method has been successfully applied to the phytochemical analysis of alkaloids extracts from two commonly used traditional Chinese herbal drugs: Sophora flavescens Ait. (Kushen) and Cortex phellodendri chinensis (Huangbai) and their medicinal preparations.     

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).     

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.     

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.     

白细胞介素-2对小鼠神经病理性痛的镇痛作用

目的探讨白细胞介素-2(IL-2)对神经病理性痛的镇痛作用及其可能的机制,为临床治疗神经病理性痛筛选新的镇痛药物奠定基础。方法通过结扎雄性C57BL/6小鼠单侧L5/L6脊神经建立神经病理性痛模型,然后利用机械刺激和冷刺激诱发的痛觉超敏实验分别观察腹腔注射不同剂量(1.0×106、2.5×106、5.0×106u/kg)IL-2对疼痛模型小鼠患侧脚掌痛阈的影响;同时,通过5.0×106u/kg IL-2注射前30 min腹腔注射纳洛酮(1 mg/kg)以观察阿片受体拮抗剂对IL-2镇痛作用的影响。结果腹腔注射IL-2可产生剂量依赖性镇痛作用,其中5.0×106u/kg和2.5×106u/kg IL-2均可显著提高模型小鼠患侧脚掌的50%缩足阈值(P<0.01)和降低5 min抬足次数(P<0.05),镇痛效应分别维持30 min和15 min,而1.0×106u/kg IL-2无明显的镇痛作用。IL-2产生的镇痛效应不能被纳洛酮阻断。结论IL-2对小鼠神经病理性痛具有镇痛作用,其镇痛效应除受阿片受体介导外还可能有其他分子参与。