盐酸洛美利嗪及有关物质的HPLC测定

采用 RP-HPL C法测定盐酸洛美利嗪的含量及其有关物质。色谱柱为 L ichrospher5 C1 8柱 (2 0 0× 4.6mm,5 μm) ,流动相为乙腈 -p H5 .8磷酸盐缓冲液 (80∶ 2 0 ) ,检测波长 2 10 nm。线性范围为 40～ 160 μg/ ml,r=0 .9999,精密度 <2 %。     

高效液相色谱法测定生长抑素含量

目的研究用高效液相色谱法测定生长抑素含量的方法。方法采用SupelcosilLC 18(4 .6mm× 15 0mm ,5 μm)色谱柱 ;以磷酸溶液 (取 85 %磷酸 11m1,加水 90 0ml,用三乙胺调pH 2 .3,加水至 10 0 0ml) 乙腈 (75∶2 5 )为流动相 ;流速 :1.0ml/min ;检测波长 :2 15nm ;柱温 :30℃ ;进样量 :2 0 μl。结果测定的线性范围为 0 .0 3～ 0 .0 7mg/min ;r=0 .9988;精密度 :RSD为 0 .0 8% (n =6 ) ;日间精密度 :RSD为 0 .5 0 % (n =5 )。结论此法简便快速、准确、专属性好     

非那雄胺有关物质及含量的RP-HPLC测定

采用反相高效液相色谱法测定了非那雄胺有关物质和含量。色谱条件 ODS C1 8色谱柱 (15 0× 4 .6 m m ,5μm) ,流动相为水 -乙腈 -四氢呋喃 (8∶ 2 .5∶ 1) ,流速 1.8m l/ min,紫外检测波长为 2 10 nm。非那雄胺含量测定的线性范围为 15～ 15 0 μg/ ml,相关系数 r=0 .9997;样品溶液在 5 d内稳定 ;日内和日间精密度均良好 (RSD<1.0 % )。     

HPLC法测定盐酸仑氨西林的含量及有关物质

目的　采用高效液相色谱法测定盐酸仑氨西林含量及有关物质。方法　 ODS- C1 8柱 (2 5 0 mm×4 .6 mm,5 μm) ,流动相为 0 .0 78mol/ L 磷酸二氢钾溶液 -乙腈 (75∶ 2 5 ) ,流速 1.0 ml/ min,柱温 4 0℃ ,检测波长2 30 nm。结果　盐酸仑氨西林在 0 .10～ 0 .4 0 mg/ ml的浓度范围内呈良好的线性关系 ,回归方程为 Y=1392 0 5 X- 15 5 1.5 (r=0 .9997) ,最低检测浓度为 0 .6 2 5 μg/ ml,平均回收率为 99.74 % (n=9) ,RSD=0 .2 4 %。结论　本方法简便、快速 ,结果准确、可靠、重现性好。     

注射用盐酸头孢吡肟中L-精氨酸的HPLC测定

采用 HPL C法测定注射用盐酸头孢吡肟中 L-精氨酸的含量。采用 L i Chrospher Diol色谱柱 (2 5 0× 4 .0 m m,5 μm) ,0 .0 1m ol/L磷酸二氢铵溶液 (用磷酸调节 p H至 2 .0± 0 .1) -乙腈 (30∶ 70 )为流动相 ,检测波长 2 0 6 nm。L-精氨酸在 5 0～ 4 0 0 μg/ml范围内线性关系良好 (r=0 .9999) ;方法的平均回收率为 10 0 .8% (RSD1.0 % )。     

高效液相色谱法测定甲磺酸帕珠沙星注射液中右旋体的含量

目的建立甲磺酸帕珠沙星注射液中右旋异构体含量的高效液相色谱测定方法。方法Shim pakCLC ODS柱 (15 0mm× 6 .0mm ,5 μm) ,流动相为L 异白氨酸硫酸铜溶液 (取L 异白氨酸 1.3g、硫酸铜 1.0g和水 10 0 0ml溶解用 0 .1mol/L盐酸或 0 .1mol/L氢氧化钠调pH至 3.5 ) 甲醇 (75∶2 5 ) ;检测波长 :32 0nm。结果甲磺酸帕珠沙星右旋体在 0 .5～ 10 0 μg/ml浓度范围内呈良好的线性关系 (r=0 .9993) ,平均回收率为 99.5 4 % ,RSD为 1.0 1% (n =9)。结论此法快捷、专属性及重现性好 ,可用于甲磺酸帕珠沙星和D 甲磺酸帕珠沙星杂质的含量测定。     

注射用甲磺酸加贝酯的反相离子对HPLC测定

建立了测定注射用甲磺酸加贝酯含量的 HPL C方法。以 Zorbax Rx- C1 8(2 5 0× 4 .6 mm,5 μm)为色谱柱 ,乙腈 -樟脑磺酸钠缓冲液 (p H 3.5 ) (5 5∶ 4 5 )为流动相 ,检测波长为 2 36 nm。甲磺酸加贝酯在 12 .4～ 2 4 8.0μg/ml浓度范围内有良好线性关系 (r=0 .9999) ,平均加样回收率为 10 0 .0 % (RSD =0 .0 6 % ) ,系统精密度为 0 .2 7%。     

高效液相色谱法测定盐酸噻氯匹定胶囊的含量

目的建立盐酸噻氯匹定胶囊的高效液相色谱 (HPLC)含量测定方法。方法盐酸噻氯匹定胶囊的HPLC含量测定色谱条件 :ODS柱 ,流动相 :0 .1%三乙胺溶液 (用磷酸调pH 2 .5 ) 甲醇 (6 0∶4 0 ) ,检测波长 :2 33nm ,流速 :1.0ml/min。结果盐酸噻氯匹定胶囊HPLC含量测定方法的线性范围为 10ng～ 10 μg ,日内RSD为0 .5 0 % ,日间RSD为 0 .4 6 %。结论此方法简便 ,准确可靠 ,耐用性好     

高效液相色谱法测定莫西沙星制剂含量

目的 :建立高效液相色谱法测定莫西沙星制剂的含量。方法 :采用InertsilODS 2色谱柱 (5 μm ,2 5 0mm× 4.0mm) ,流动相为 1%三乙胺溶液 (磷酸调pH值至 4.5 )—乙腈 (84∶16,v/v) ;柱温 :40℃ ;检测波长 :2 96nm ;流速 :1.0ml/min。结果 :线性范围 0 .4～ 3 μg ,r =0 .9999,平均回收率为 10 0 .7% (RSD =0 .7% )。结论 :方法简单 ,专属性好 ,可用于莫西沙星制剂的含量测定。     

反相离子对高效液相色谱梯度洗脱测定复方硝酸益康唑乳膏含量

目的　建立复方硝酸益康唑乳膏含量测定的方法。方法　色谱柱为辛烷基硅烷键合硅胶柱(L i Chrospher C8柱 ,5 μm,4 .6 mm× 2 5 0 mm) ,检测波长 2 2 7nm,采用 A相 (85 %磷酸溶液 1ml,加无水己烷磺酸钠 0 .94 g,溶于 72 0 ml水中 ,加乙腈 14 0 ml,异丙醇 14 0 ml) - B相 (85 %磷酸溶液 1ml,加无水己烷磺酸钠0 .94 g,溶于 10 0 ml水中 ,加甲醇 90 0 ml)为流动相进行线性梯度洗脱 (A相起始比例为 10 0 % ,2 5 min内 B相上升为 10 0 % ,保持该比例至硝酸益康唑完全洗脱 ) ,流速为 1ml/ min,柱温为 4 0℃ ,进样量为 2 0 μl,内标物质为对羟基苯甲酸乙酯。结果　硝酸益康唑在 0 .3～ 0 .8mg/ ml,曲安奈德在 0 .3～ 0 .8mg/ ml,苯甲酸在 0 .0 6～0 .16 mg/ ml范围内 ,浓度与其峰面积均呈良好线性关系 (r=0 .9999,n=6 ) ,平均回收率分别为硝酸益康唑10 1.5 % ,曲安奈德 10 0 .3% ,苯甲酸 98.8%。结论　本法快速、准确 ,可同时测定复方硝酸益康唑乳膏中硝酸益康唑、曲安奈德、苯甲酸的含量。     

Acute toxicity of polyethylene glycol p-isooctylphenol ether in Syrian hamsters exposed by inhalation or bronchopulmonary lavage

Dose-response studies were conducted with Syrian hamsters exposed to polyethylene glycol p-isooctylphenyl ether (Triton X-100) via inhalation or bronchopulmonary lavage. Syrian hamsters were exposed to an aerosol of Triton X-100 with a mass median aerodynamic diameter of 1.5 μm and a concentration of 3.0 mg/liter. Estimated initial lung burdens of Triton X-100 ranged from 800 to 3100 μg. Hamsters were lavaged with concentrations of Triton X-100 ranging from 0.01 to 0.10% in isotonic saline resulting in initial lung burdens of Triton X-100 that ranged from 300 to 3200 μg. The $\text{LD}\text{50}\text{7}$ values were 1700 μg (1300–2100 μg, 95% confidence limits) for the inhalation study and 2100 (1900–2700) μg for the lavage study. The difference between the $\text{LD}\text{50}\text{7}$ values for the two methods of exposure was not significant. However histopathological examination revealed differences in the nature and distribution of pathologic changes observed in animals exposed by the two routes of administration. Animals exposed by inhalation died as a result of ulcerative laryngitis and laryngeal edema with only minimal pulmonary pathologic alterations. Animals exposed by lavage, where the larynx was not exposed to Triton X-100, died from pulmonary edema and acute exudative pneumonia, these results demonstrate the need for careful selection of exposure methods to meet the specific objectives of a toxicology study.     

金黄色葡萄球菌α-毒素溶血活性测定方法优化及初步应用

 目的  优化金黄色葡萄球菌α-毒素（α-toxin,AT）溶血活性测定方法,用于无毒突变AT（non-toxic mutant AT,mAT）特异性血清抗体体外功能性评价和中和抗体滴度测定。方法  使用AT裂解兔红细胞,释放血红蛋白,通过分光光度法检测溶血活性,并对该方法中的孵育时间、兔红细胞终浓度、Triton X-100浓度进行优化,同时验证方法重复性。对mAT特异性血清抗体进行体外功能性评价,绘制AT溶血曲线,计算AT 溶血率50% 时的浓度,检测血清半数中和抗体滴度。结果  AT溶血活性最适检测条件为：孵育时间90 min,兔红细胞终浓度为2%（V/V）,Triton X-100浓度为0.25%。此条件下,分别加入mAT和含mAT的多价抗原血清抗体,对AT溶血活性均起到明显的抑制作用。AT 溶血率50%时浓度为1.75 μg/ml,变异系数3.75%。mAT和含mAT的多价抗原血清抗体的半数中和抗体滴度初步结果均为1∶64。结论  优化的AT溶血活性检测方法重复性良好,且检测时间较短,可用于特异性血清抗体体外功能性评价和中和抗体滴度测定。     

不同条件对RiboGreen法检测mRNA疫苗中mRNA含量的影响

目的 探讨RiboGreen法检测mRNA疫苗中mRNA含量可能的影响因素。方法 在不同裂解液浓度、裂解时间、样品稀释浓度和参比品条件下，使用RiboGreen荧光染料法对不同生产工艺的mRNA疫苗的mRNA含量进行检测。结果 在裂解液为0.1%〜2.5% Triton X-100、裂解5〜30 min、样品稀释至mRNA含量0.5〜1.5 μg/ml时，mRNA含量检测结果的差异无统计学意义；不同参比品序列下样品mRNA含量检测结果差异有统计学意义。结论 根据检测样品的mRNA含量的回收率，初步确定了mRNA疫苗中mRNA含量检测的最适条件。     

Influence of elastase-induced emphysema and the inhalation of an irritant aerosol on deposition and retention of an inhaled insoluble aerosol in Fischer-344 rats

The purpose of this study was to assess the effects of elastase-induced pulmonary emphysema and the inhalation of an irritant aerosol (Triton X-100, a nonionic surfactant similar to those used in a number of pressurized consumer products) on pulmonary deposition and retention of an insoluble test aerosol, 59Fe-labeled Fe2O3. Untreated rats or rats pretreated by intratracheal instillation with elastase were exposed to an aerosol of 59Fe-labeled Fe2O3 either 18 hr or 7 days after exposure to aerosolized Triton X-100 which was administered in doses of 20, 100, or 200 micrograms/g of lung. Rats pretreated with elastase had significantly lower pulmonary deposition of 59Fe than the untreated controls (p less than 0.005). Pulmonary deposition of Fe2O3 was unaffected by pretreatment with Triton X-100. Elastase treatment alone had no effect on retention of Fe2O3. Triton X-100 administered 18 hr prior to exposure of rats to Fe2O3 aerosol resulted in dose-related increases in whole-body retention of 59Fe. When rats were exposed to Triton X-100 7 days before exposure to Fe2O3, increased retention of 59Fe was noted only in those treated at the highest Triton X-100 dose level (200 micrograms/g).     

Method development and validation for the GC-FID assay of tributyl phosphate in a phospholipid emulsion

This paper describes the development and validation of an isothermal GC-FID method for the assay of tributyl phosphate in a phospholipid emulsion. The emulsion is used as a topical ointment to deliver Triton X-100, a spermicide. The tributyl phosphate is added to the emulsion as a plasticizer or softening agent. The chromatographic conditions of the method employ a J&W DB-Wax capillary column (30 m x 0.53 mm, film thickness 1 microm), isothermal elution with He at a column flow of 2.0 ml/min, injector, detector, and oven temperatures at 210 degrees C, a split ratio of 18.0/2.0, and a 3-microl injection volume. Sample calibration was performed with tributyl phosphate purchased from Aldrich (USP Reference Standard is not available). The linearity of the tributyl phosphate peak area responses was demonstrated from approximately 50 to 150% of the analytical concentration of 100 microg/ml. System precision was determined from five replicate injections of a standard and sample solution. Reproducibility of the tributyl phosphate peak area responses showed R.S.D. of 1.2 and 0.4%, respectively. Method precision was performed by assaying five samples by two different analysts on different days. The mean %LC was 95.5% (R.S.D.=1.0%) for the first analyst, and 95.6% (R.S.D.=1.0%) for the second analyst. The mean %LC value for all ten sample preparations was 95.5% (R.S.D.=0.9%). The limits of detection and quantitation were determined to be 0.2 and 0.7 microg/ml, respectively.     

Development and validation of a HPLC method for routine quantification of the decapeptide Cetrorelix in liposome dispersions

The development and validation of an HPLC method for the quantification of the decapeptide Cetrorelix (acetyl-D-2-naphthylalanyl-D-4-chlorphenylalanyl-D-3-pyridylalanyl-seryl-tyrosyl-D-citrullyl-leucyl-arginyl-prolyl-d-alaninamide), a potent antagonist of the luteinising hormone-releasing hormone in liposome dispersions is described. An isocratic reversed phase method with UV-detection appeared most appropriate. Several detergents were tried to disrupt liposomes. Furthermore, detergents turned out to be useful, because they minimised unwanted loss of Cetrorelix due to adsorption to the vial surfaces. Triton X-100 was found most effective, while sodium cholate led to quantification problems. In the presence of 2.5% Triton X-100 calibration curves with a high degree of linearity were achieved in the desired range of 0.2-10 microg/ml. The limits of detection and quantification of Cetrorelix were calculated from the peak-to-noise ratio to be 11 and 37 ng/ml, respectively. The repeatability of the method in presence of phospholipid and Triton was good with relative standard deviations (R.S.D.) ranging from 0.8% (at 0.05 microg/ml) to 1.5% (at 0.2 microg/ml). The presence of liposomes at phospholipid contents of up to 0.25mg/ml did not significantly affect the slope or linearity of the calibration curve, nor the peak-to-noise ratio.     

Morphological and physiological changes in Tetrahymena pyriformis for the in vitro cytotoxicity assessment of Triton X-100.

Non-ionic surfactants such as Triton X-100 have been widely used in industrial processing and in cleaning products for almost 50 years, being effective and economic emulsifying, wetting agents, dispersants and solubilizers. Cleaning products containing these surfactants are disposed of mainly by discharge into wastewater, which receives biological treatment in wastewater treatment systems. However, surface-active agents interact with eukaryotic cell membranes leading to biological damage at high concentrations. Tetrahymena pyriformis was used here as model organism to assess the effects of Triton X-100 through a series of in vitro cytotoxicity tests. Growth rates and morphological changes were, by their simplicity and reproducibility, the simplest toxicological assays. Cytoskeleton analysis seemed to be related with phagocytosis rate. Viability was evaluated by two different tests. Calcein AM/EthD-1 was used to assess T. pyriformis membrane damage during the 48-h experiment. The colorimetric MTT assay proved to be highly sensitive even at very short periods of Triton X-100 exposure. Tests performed in this study included simple and fast bioassays that provide overall information on the morphological and physiological state of cells exposed to different non-lytic and lytic concentrations of Triton X-100.     

The mechanism of action of sanguinarine against methicillin-resistant Staphylococcus aureus

Sanguinarine is a benzophenanthridine alkaloid derived from the root of Sanguinaria canadensis. It is known to perform a wide spectrum of biological activities. The aim of this study is to examine the antimicrobial actions of sanguinarine against methicillin-resistant Staphylococcus aureus (MRSA). Sanguinarine antimicrobial activity was assessed by broth dilution method; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors and transmission electron microscopy were used to monitor the survival characteristics and the changes in bacteria morphology. The activity of sanguinarine against MRSA strains ranged from 3.12 to 6.25 μg/ml, while the minimum inhibitory concentrations of the two reference strains are 3.12 μg/ml and 1.56 μg/ml. The treatment of the cells with sanguinarine induced the release of membrane-bound cell wall autolytic enzymes, which eventually resulted in lysis of the cell. The OD(600s) of the suspensions treated with the combination of Tris-(hydroxymethyl) aminomethane and Triton X-100 with sanguinarine were reduced to 40% and 8%, respectively. Transmission electron microsco-py of MRSA treated with sanguinarine showed alterations in septa formation. The predisposition of lysis and the altered morphology seen by transmission electron microscopy suggest that sanguinarine compromises the cytoplasmic membrane.     

顶空气相色谱法测定猪纤维蛋白原中的乙醇残留量

目的 建立顶空气相色谱法测定猪纤维蛋白原中的乙醇残留量,并对方法进行验证和初步应用.方法 采用Agilent DB-624毛细管柱,以高纯氮气为载气,顶空进样,由氢火焰离子化检测器检测信号,通过内标法计算乙醇含量.确定标准曲线线性范围以及检测限和定量限,并对方法的专属性、准确性、重复性、稳定性、耐用性进行验证.应用该方法对3批猪纤维蛋白原样品中的乙醇残留量进行检测.结果 空白基质、供试品、对照(乙醇)及内标物(正丙醇)在保留时间处相互均无干扰,且对照和内标物峰形良好.标准曲线的线性范围为5.390～80.850 μg/ml.检测限为1.617ug/ml,定量限为5.390 μg/ml.低、中、高3个质量浓度乙醇的总平均加标回收率为(102.84±3.57)％,总相对标准偏差为3.00％;重复检测6次的相对标准偏差为2.17％.对照品和供试品在不同时间点与起始时间的乙醇含量之比为99.74％～105.26％.方法参数微小变动前后测得样品中的乙醇含量之比为94.00％～100.18％.3批猪纤维蛋白原样品中的乙醇残留量分别为35、36、54 μg/ml,均满足中国药典2015年版三部的限量要求(不高于250 μg/ml).结论 本法专属性强,准确性、重复性、稳定性和耐用性均良好,可用于猪纤维蛋白原中的乙醇残留量检测.     

HPLC法同时测定多维营养素片中维生素B6、烟酰胺、咖啡因的含量

目的优选HPLC法同时测定多维营养素片中维生素民、烟酰胺、咖啡因的检测方法。方法样品以甲醇：水：磷酸=100：400：0．5的混合溶液为溶剂,超声处理5min,色谱柱：Wondasil C18（250mm×4．6mm,5μm）;流动相：0．04％戊烷磺酸钠溶液（用冰醋酸调节pH值至3．O）-甲醇（82：18）;流速为1．0ml／min;检测波长：280nln。结果维生素B6的线性范围为0．5～4．0斗g,相关系数0．9997,平均回收率99．8％,检出限0．01764μg／ml;烟酰胺的线性范围0．5—4．0μg,相关系数0．9999,平均回收率101．3％,检出限0．9178μg,／ml;咖啡因的线性范围0．2～2．0μg,相关系数1．0000,平均回收率99．O％,检出限0．01561μg／ml。结论该方法灵敏度高,选择性好,分离效果好、线性关系及重现性好,适合于同时测定多维营养素类保健品中维生素B6、烟酰胺、咖啡因的含量。