注射用头孢哌酮钠舒巴坦钠含量测定方法的改进

注射用头孢哌酮钠舒巴坦钠收载于中国药典2005年岸縖1],含量测定项采用HPLC法测定头孢哌酮和舒巴坦的含量.笔者在试验过程中发现:采用药典方法测定头孢哌酮的含量,头孢哌酮峰不仅保留时间长,而且有拖尾现象,通过改变流动相比例并加柱温后,头孢哌酮峰的保留时间缩短且无拖尾现象,对二者的含量测定结果均无影响,极大地缩短了试验时间,提高了工作效率.     

RP-HPLC法测定罗红霉素片中罗红霉素的含量

目的建立罗红霉素片含量测定的高效液相色谱法,并与微生物检定法、紫外分光光度法的测定结果进行比较。方法采用Agilent-C18色谱柱（4.6 mm×250 mm,5μm）;0.067 mol·L^-1磷酸二氢铵（三乙胺调节pH6.5）-乙腈（3∶2）,加入1%的三乙胺,用磷酸调节pH7.2为流动相;流速1.0 ml·min^-1;检测波长211 nm;柱温30℃。结果罗红霉素在2.14～42.80 mg·L^-1的浓度范围内有良好的线性关系（r=0.999 9）,平均回收率为97.8%,RSD为0.78%（n=5）。结论三种含量测定方法的结果相近,均可用于罗红霉素片的含量测定。     

HPLC法测定罗红霉素颗粒的含量

罗红霉素为红霉素的衍生物,中国药典2000年版已收载,采用微生物效价法测定罗红霉素的含量.国内文献报道罗红霉素及其制剂含量测定采用高效液相色谱法,均为C18柱,以醋酸铵缓冲液与溶剂(乙腈或甲醇)做流动相,在210～215 nm处检测,但其色谱峰与有关物质不能很好的分离.笔者采用的HPLC系统,在室温条件下等度洗脱,就可以将有关物质与罗红霉素很好的分离.     

地塞米松磷酸钠注射液质量检查方法改进

目的:改进地塞米松磷酸钠注射液含量测定方法并建立杂质检查方法.方法:采用高效液相色谱外标法测定含量及检查杂质.流动相为甲醇-0.34%磷酸二氢钾溶液(60:40);色谱柱为Nova-pak C18柱;检测波长为240 nm.结果:外标法测定含量与内标法(药典法)比较,结果一致(P＞0.05).不同生产厂家的产品杂质含量存在较大差异,确定其杂质峰面积与总峰面积之比不得大于5%.结论:外标法含量测定方法简便快捷;杂质检查直观、可靠.     

RP-HPLC法测定甲氧氯普胺片中甲氧氯普胺的含量

目的建立高效液相色谱测定甲氧氯普胺片中甲氧氯普胺的含量。方法采用高效液相色谱法[色谱柱:依利特,BDS(5μm,4.6mm×250mm);流动相:甲醇-乙腈-醋酸盐缓冲液(pH4,含0.1%三乙胺)(7∶18∶75);流速:1.0ml.min-1;检测波长:273nm;柱温:40℃。结果实验表明甲氧氯普胺含量在18.4~73.6μg.mL-1范围内与峰面积呈良好线性关系(r=0.9999);加样回收率考察,甲氧氯普胺平均回收率为99.4%,RSD=1.2%(n=9)。结论本方法准确可靠,可作为甲氧氯普胺片的含量测定方法。     

高效液相色谱法测定罗红霉素胶囊中罗红霉素的质量

目的:建立罗红霉素胶囊中罗红霉素质量的高效液相色谱法测定方法。方法:采用ODSC18柱(250mm×6mm,5μm),以0.1mol/L磷酸二氢铵溶液(三乙胺调节pH6.5)-乙腈(65∶35)为流动相,流速为1.0mL/min,检测波长210nm,进样量20μL。结果:罗红霉素在质量浓度10～80μg/mL的范围内与峰面积响应值呈良好线性关系(r=0.9997,n=5),平均回收率为99.84%,RSD=0.65%(n=6)。结论:该法操作简便,数据准确、可靠,适用于罗红霉素胶囊中罗红霉素质量的控制。     

D-氨基葡萄糖硫酸钾盐含量测定方法探讨

目的 对D-氨基葡萄糖硫酸钾盐的含量测定方法的适用性进行验证.方法 采用Zorbax C8(4.6 mm×250 mm,5 μm)色谱柱,流动相为乙腈-0.05%磷酸(pH=3.0)(40:60),流速0.6 mL·min-1,检测波长195 nm.结果 D-氨基葡萄糖盐酸盐对照品和D-氨基葡萄糖硫酸钾盐样品图谱中的主峰可能不是D-氨基葡萄糖峰,而是Cl-峰; 在药典规定的色谱条件下,D-氨基葡萄糖硫酸钾盐中的SO42-峰与主峰Cl-峰重叠,干扰主峰的测定.结论 需进一步改进和完善D-氨基葡萄糖硫酸钾盐的含量测定方法.     

HPLC法测定阿莫西林钠含量探讨

本文对《中国药典》2000年版和《英国药典》2000年版中阿莫西林钠的高效液相测定方法进行了对比研究。色谱柱:天和色谱液相填充柱C_(18)(250mm×4.6mm 10μm);流动相:水-甲醇(75:25),pH5.0磷酸缓冲液-乙腈(96:4);检测波长:254nm。在0.1～O.5mg·ml~(-1)范围内峰面积与浓度呈良好的线性关系,r=0.9996,以pH5.0的磷酸缓冲液为溶剂。本方法在阿莫西林钠含量测定过程中具有良好的稳定性及重现性。     

HPLC法测定枸橼酸西地那非阴道栓的含量

目的 建立枸橼酸西地那非阴道栓的含量测定方法,以控制其产品质量.方法 采用高效液相色谱测定法.C18色谱柱.流动相为:0.05mol·L-1磷酸三乙胺(0.7%三乙胺1000mL用磷酸调节pH3.0)甲醇-乙腈(52 ∶ 30∶18),检测波长290nm.结果 在10μg～60μg·mL-1范围内峰面积与西地那非浓度成良好的线形关系,加样回收率和RSD分别为99.0%和0.6%.结论 方法简便准确,可用于制剂的质量分析.     

RP-HPLC测定妇康洗剂中苦参碱的含量

目的建立妇康洗剂中苦参碱的含量RP-HPLC测定方法.方法色谱柱Kromsail C18柱;流动相乙腈-0.02三乙胺溶液(3070);流速1.0 ml·min-1;检测波长220 nm.结果苦参碱在0.624～6.240μg范围内与峰面积成良好的线性关系(r=0.999 3);平均加样回收率为98.3%,RSD=1.25%(n=5).结论本法灵敏、准确,重现性好,可作为妇康洗荆中苦参碱的含量测定.     

Application of Multidimensional Screening and Analysis and Computer Simulation Software for Rapid HPLC Method Development

A comprehensive automated strategy for the development of a reversed-phase high-performance liquid chromatography-UV chromatographic method for a basic drug candidate (pKa 7.7), its impurities, and degradants was demonstrated by using multidimensional screening and analysis (MeDuSA) and Drylab^® computer optimization software. Phase 1 of the strategy employed an automated column selection system and solvent screening (MeDuSA). Samples were screened using ten columns of varying selectivity and high pH stability in combination with four mobile phases of differing pH (pH 2, 4, 7, and 10) containing acetonitrile and methanol as organic modifiers. The best chromatographic conditions (greater number of resolved impurities, tailing factor ~1.0, resolution >1.4) were obtained using the methyl hybrid organic–inorganic polymer phase Xbridge C18 and pH 7 and pH 10 ammonium acetate/methanol/acetonitrile mobile phase. Further screening in MeDuSA using mobile phases A: 0.01 M NH₄OAc in CH₃CN/water (5:95), and B: 0.01 M NH₄OAc in CH₃CN/water (95:5) at pH 7 demonstrated that CH₃CN is the more critical organic modifier for selectivity. In phase 2, DryLab^® software was used to model the effect of temperature, pH, and ionic strength of the buffer on resolution and peak shape to determine the design space of the method and establish the optimal operating conditions. In silico optimization supported by DryLab^® enabled the determination of the optimum conditions without carrying out trial-and-error simulations. Excellent agreement between Drylab^® simulation and experimental results was obtained. As a result of this strategy, a method with the highest resolution of all components, optimum tailing factor (~1.05), and decreased run time (from 35 to 14 min compared to the original method) was developed faster and with higher efficiency in comparison to the traditional method development process.     

Determination of naphthidrofuryl and naphthidrofurylic acid in human plasma using RP-HPLC and fluorimetric detection

An analytical method was developed for the determination of naphthidrofuryl and its principle metabolite--naphthidrofurylic acid in human plasma by the RP-HPLC method with fluorimetric detection. The analytical method is based on a single extraction with a 5 ml mixture ether: hexane (1:1 by volume), with salting out by means of potassium chloride after which the organic phase after centrifugation, evaporation and reconstitution is sprayed on the reverse phase of HPLC and the separated substances are determined fluorimetrically (excitation 271 nm, emission 240 nm). Lonazolac served as the internal standard. The mobile phase consisted of 72% methanol, 1% triethylamine, 0.6% phosphoric acid. Optimization of the composition of the mobile phase from the aspect of the amino base, dependence of the percent content of methanol in the mobile phase and dependence of the recovery of substances on the composition of the extracting reagent are presented. The limit of detection was 4 ng/ml of plasma for naphthidrofuryl. The stability of compounds (a minimum of 2 months) and interference of some other drugs are shown.     

Validation of a Stability-Indicating RP-HPLC Method for the Simultaneous Determination of Trimethoprim and Sulfadimethoxine Sodium in Oral Liquid Dosage Form

A simple, specific, accurate, and stability-indicating RP-HPLC method was developed and validated for the simultaneous determination of Trimethoprim (TMP) and Sulfadimethoxine sodium (SDMS) in Vetricine^® oral solution product. The desired separation was achieved on an ODS column (250×4.6 mm i.d., 5 μm) at room temperature. The optimized mobile phase consisted of an isocratic solvent mixture of water:acetonitrile:triethylamine (700:299:1, v/v/v), adjusted to a pH of 5.7 ± 0.05 with 0.2N acetic acid. The mobile phase was fixed at 0.8 ml/min and the analytes were monitored at 254 nm using a photodiode array detector. The effects of the chromatographic conditions on the peaks USP tailing factor, column efficiency, and resolution were systematically optimized. Forced degradation experiments were carried out by exposing TMP, SDMS standards, and the oral solution formulation to thermal, photolytic, oxidative, and acid-base hydrolytic stress conditions. The degradation products were well-resolved from the main peaks and the excipients, thus proving the reliable stability-indicating method. The method was validated as per ICH and USP guidelines (USP34/NF29) and found to be adequate for the routine quantitative estimation of TMP and SDMS in commercially available Vetricine® oral liquid dosage form.     

中国药典“利福平胶囊”含量测定方法的修改建议

目的:改善含量测定中利福平溶液的稳定性.方法:分别以药典方法的流动相、甲醇、乙腈、缓冲液为溶剂考察溶液稳定性.结果:按中国药典2000年版方法,利福平溶液1小时约降解20%.改以甲醇-0.01mol/L磷酸二氢钾溶液(pH值为6.5)(1:1)为溶剂为宜.结论:利福平在药典方法的流动相中极不稳定,但适当改变溶剂可以增加其稳定性,建议立即修改药典.     

A new universal high-performance liquid chromatographic method for determination of 1,4-benzodiazepines as bulk drugs and in pharmaceutical formulations

A universal high-performance liquid chromatographic method was developed for the simultaneous determination of seven frequently prescribed 1,4-benzodiazepines in bulk powder or formulated in tablets or capsules. Peak tailing commonly reported with HPLC method developed for benzodiazepines, is completely omitted with this new method. The assay procedure consisted of pulverization, extraction into methanol, filtering, diluting and injecting of aliquots of clear filtrate into a reversed phase column with a very low silanol activity. The mobile phase consisted of a mixture of methanol and 0.05 mol x L(-1) buffer solution of ammonium dihydrogen phosphate (50:50, v/v) adjusted to pH 5.8. The effluent was monitored by UV detection at 254 nm and delivered at a flow rate of 1.6 mL x min(-1). The new method has been applied for quantifying of 1,4-benzodiazepines in different commercial formulations with recoveries ranging from 99.0 (+/- 1.98) to 102.0 (+/- 1.58)%. The excipients present in tablets and capsules did not interfere with the developed method. The applicability of the method for content uniformity and dissolution tests has been also investigated and the results were considered satisfactory. The developed method is rapid and sensitive, and is suitable for routine control of pharmaceutical dosage forms.     

HPLC法测定双氯芬酸钠肠溶片的含量及含量均匀度

目的:建立了高效液相色谱法测定双氯芬酸钠肠溶片中双氯芬酸钠的含量及含量均匀度的方法。方法:色谱柱为KromasilC18(250mm×4.6mm,5μm),流动相为甲醇-水(75:25),用磷酸调节pH至3.50±0.05,检测波长为284nm,流速为1ml?min-1。该方法对双氯芬酸钠的平均回收率为99.40%,RSD为1.24%(n=5)。该法与与标准法比较,没有显著性差别。结果:方法简便,结果准确,专属性强。结论:可用于含量及含量均匀度的测定。     

HPLC法测定沙纳唑的含量及其有关物质

目的:建立高效液相色谱法测定沙纳唑的含量及其有关物质。方法:采用 Kromasil KR 100 C_(18)色谱柱(4.6 mm×250mm,5μm)。流动相:甲醇-水-冰醋酸(200:800:3),流速:1 mL·min~(-1)。检测波长:248 nm。结果:HPLC法测定的线性范围为50～300μg·mL~(-1),r=0.9999,最低检测限为0.5 ng,本方法重复性和精密度良好(RSD<2％)。结论:采用HPLC法测定沙纳唑及其注射液的含量及有关物质,方法简便,结果准确。     

高效液相色谱法测定兔血清中丁螺环酮的浓度

目的 :建立高效液相色谱法测定兔血清中丁螺环酮浓度的方法。方法 :采用ZORBAXSB -C18 色谱柱 ,以甲醇 -乙腈 -水 -0 1mol/L醋酸钠缓冲液 (pH4 5) (10∶38∶51∶1)为流动相 ,检测波长为237nm。以峰面积内标法测定浓度。结果 :本方法能有效分离被测组分 ,其丁螺环酮在1～80ng/ml浓度范围内与峰面积呈现良好的线性关系 ,r=0 9979。结论 :本方法操作简便 ,测定结果准确 ,灵敏度高 ,适用于血清药物浓度测定     

超快速液相色谱法测定新型“K2”香料中JWH-018的含量

目的:建立测定新型"K2"香料中JWH-018含量的超快速液相色谱法。方法:采用Shim-pack XR-ODSⅡ色谱柱(2.0 mm×75 mm,2.2μm);流动相:乙腈(含0.1%甲酸)-水(含0.1%甲酸)(80:20,v/v),流速:0.25 mL·min-1;检测波长280 nm,柱温:35℃,进样量:2μL,外标法定量测定JWH-018的含量。结果:JWH-018的浓度在1-1000μg·mL-1范围内线性良好(A=8419.8C+13932,r=1.0000);最低检测限为90 ng·mL-1(S/N=3);低、中、高3种浓度的加样回收率(n=3)分别为101.0%(RSD=0.28%),101.0%(RSD=0.63%),101.7%(RSD=0.03%)。结论:本法操作简便、灵敏、准确,重复性好,适用于"K2"香料中JWH-018含量的测定。     

柱前衍生化HPLC法测定齿瓣石斛中总氨基酸的含量

目的:建立测定齿瓣石斛中总氨基酸含量的方法。方法:以2,4-二硝基氟苯为衍生试剂,与齿瓣石斛中氨基酸柱前衍生,采用高效液相色谱法测定其含量。色谱柱为SymmetryC1(8250mm×4.6mm,5μm),流动相A为0.05mol·L-1醋酸钠-醋酸缓冲液(pH6.4)-1%N,N-二甲基甲酰胺,流动相B为乙腈-水(1:1,V/V),梯度洗脱,检测波长为360nm,柱温为30℃,进样量为20μL。结果:13种氨基酸的线性相关系数均>0.9990,平均加样回收率范围在99.34%～101.93%之间,RSD在0.54%～1.91%之间(n=6)。结论:本方法快速、简便、结果准确,可为齿瓣石斛的深入研究奠定基础。