分离测定利伐沙班及其杂质的方法及应用

本发明属于分析化学领域,具体涉及一种分离测定利伐沙班及其杂质的方法及应用。用于分离测定利伐沙班及其杂质的试剂组合物,稀释剂:乙腈酸的水溶液;流动相A:含有甲醇和乙腈的缓冲盐溶液;流动相B:缓冲盐溶液和乙腈的混合液。采用上述试剂组合物分离测定利伐沙班及其杂质的方法:取利伐沙班溶于稀释剂得样品溶液;配制流动相;将样品溶液注入分离检测系统中,流动相对样品溶液进行洗脱分离得洗脱液;洗脱液进入检测器测定。本发明能将利伐沙班及其杂质完全分离,基线好,灵敏度高,分析时间短,重现性好,可以有效分离及测定利伐沙班原料药和制剂中各有关物质含量。     

高效液相色谱法测定培美曲塞二钠原料药中有关物质

建立了高效液相色谱法测定培美曲塞二钠原料药有关物质的方法。采用Zorbax SB-C8(4.6 mm i.d.×150 mm,3.5μm)色谱柱,以0.025 mol/L乙酸钠溶液(用冰乙酸调p H至5.5)为流动相A,乙腈为流动相B,梯度洗脱,流速为1.0 m L/min;柱温35℃;检测波长250 nm;进样体积20μL。在该条件下主成分与有关物质的分离度良好,在测定的范围内具有良好的线性关系(相关系数r0.999)、精密度和稳定性。方法能够对培美曲塞二钠原料药的主要杂质进行有效的分离和检测,可用于该产品的质量控制。     

磺胺二甲嘧啶及有关物质HPLC检测方法验证

建立了磺胺二甲嘧啶及其可能存在的5种杂质成分:对氨基苯磺酸、4,6-二甲基-2-羧基嘧啶、磺胺脒、磺胺和2-氨基-4,6-二甲基嘧啶的高效液相色谱分离和定量分析方法.采用C18色谱柱,pH 4.0的乙腈-醋酸(体积比1:9)流动相,分离有关杂质,而水-乙腈-冰醋酸(体积比87:12:1)流动相用于主成分磺胺二甲嘧啶(SM2)含量的测定;二极管阵列检测器,检测波长275 nm.上述5种杂质的检出限分别为0.023、0.023、0.009、0.009、0.030 mg/L,满足测定要求.     

辛硫磷和氯氰菊酯复配制剂的高效液相色谱分析

提出了同时测定辛硫磷和（高效）氯氰菊酯含量的主同效液相色谱分析方法。采用正相柱、２３５ｎｍ紫外检测，石油醚／乙腈流动相系统实现辛硫磷及其杂质与（高效）氯氰菊酯异构体的良好分离。     

高效液相色谱法测定西咪替丁片有关物质

建立高效液相色谱测定方法测定西咪替丁片有关物质包括西咪替丁及6种西咪替丁杂质。选择Welch XtimateTMC18色谱柱（250 mm×4.6 mm,5μm）为分离柱;以甲醇–水溶液（0.4 mL三乙胺、0.86 g的己烷磺酸钠加水780mL混合后用磷酸调pH 2.8）（体积比为24∶78）为流动相A,甲醇为流动相B,按梯度洗脱;流量为1.0 mL/min;柱温为30℃;检测波长为220 nm。西咪替丁及6种西咪替丁杂质色谱峰面积与质量浓度在10～200μg/mL内呈良好的线性关系,相关系数均大于0.999 8;方法检出限为0.022 5～1.742μg/mL。辅料空白加入3种浓度水平时7种化合物的平均回收率为97.3%～102.2%,测定结果的相对标准偏差为0.92%～3.32%(n=9)。该方法稳定可靠,适用于西咪替丁片有关物质的测定。     

高效液相色谱法测定造影剂碘普罗胺中的相关杂质

提出采用高效液相色谱法测定一种非离子型碘造影剂碘普罗胺中5种杂质含量的方法.以Eclipse C18 Plus色谱柱为分离柱,以不同体积比混合的水和体积比为55:40:5的水-乙腈-2-丁醇混合溶液为流动相进行梯度洗脱,在检测波长245 nm处对5.0 g·L-1的样品溶液中的5种杂质进行测定.结果显示:杂质1～5标准...     

高效液相色谱法测定头孢他啶的含量及杂质

采用高效液相色谱法测定了头孢他啶的含量及杂质。以Alltima C18色谱柱(250 mm×4.6 mm,5 μm)为分离柱,以乙腈和磷酸盐缓冲溶液(pH 3.9)分别为流动相A和流动相B进行梯度洗脱,流速1.3 mL/min,柱温35 ℃,紫外检测波长255 nm。从头孢他啶药物中共分出14个杂质,且14个杂质间具有良好的分离度。头孢他啶在0.267~1069 μg/mL范围内与峰面积具有良好的线性关系(r=1.0000);其定量限(S/N=10)和最低检出限(S/N=3)分别为3.1 ng和0.93 ng。3个浓度的日内测定值的相对标准偏差(RSD）为0.72%(n=3),日间测定值的RSD为0.91%(n=3)。头孢他啶溶液在4 ℃避光条件下放置24 h保持稳定。本方法与欧洲/英国药典和日本药局方的方法比较,具有分离杂质数量多、分离度好的优点。     

铜/金修饰电极-阴离子交换色谱-直流安培法测定阿米卡星注射液组分

建立了一种用阴离子交换色谱分离、以自制铜/金修饰电极为工作电极的直流安培电化学法(DC)直接检测硫酸阿米卡星注射液中主要组分及杂质含量的分析方法。考察了流动相浓度、测定电位等参数对色谱分离和测定的影响。在固定相为CarboPacPA10阴离子交换柱、流动相为26 mmol/L NaOH的色谱条件下,检测电位为0.64 V时,阿米卡星在0.0005~0.02 g/L(r=0.9989)和0.02~0.2 g/L(r=0.9991)两个浓度范围内呈线性。与裸Au电极在采用脉冲安培检测模式(PAD)时相比,电化学检测所需要的碱性强度低(pH<13),而且测定灵敏度高,线性范围宽。本方法不需要柱前和柱后衍生化,能同时测定硫酸阿米卡星注射液中的主要组分和杂质组分。修饰电极制作方法简单,催化稳定性好,可望被应用到流动注射、毛细管电泳等其它流动体系中,对硫酸阿米卡星原料药、注射液等实际样品中的各组分进行测定。     

反相高效液相色谱法同时测定丁酸氯维地平中10种相关杂质

建立反相高效液相色谱辅-光电二极管阵列检测器(RP-HPLC)法同时测定丁酸氯维地平原料药中的10种杂质。色谱柱为Symmetry C18柱(250 mm×4.6 mm,5μm),流动相为0.05 mol/L Na H2PO4溶液(pH 2.5)-乙腈/甲醇(3∶2,V/V),梯度洗脱,柱温35℃,流速为1.5 m L/min,检测波长220 nm。丁酸氯维地平及其10个已知杂质能够达到良好的分离,且各组分在各自测定浓度范围内与峰面积的线性关系良好(r≥0.9970);丁酸氯维地平及杂质1~10的检出限(S/N=3)在0.15~0.90 mg/L之间。本方法快速、简便、有效,可用于丁酸氯维地平原料药的质量控制管理。     

超高效凝胶色谱法测定聚乙二醇衍生物的分子量及其分布

采用超高效聚合物色谱(APC)技术,以单甲氧基聚乙二醇丙醛(m PEG_p ALD)为代表,测定了聚乙二醇衍生物的相对分子质量及其分布和杂质含量,优选了色谱柱和流动相,考察了样品质量浓度变化以及溶解时间等对测定结果的影响。优化后3根超高效凝胶色谱柱串联,在柱温40℃,流动相95%甲醇,流速0.5m L/min,示差折光检测条件下,对m PEG_p ALD的分子量及其分布进行测定,同时得到杂质的相对含量。结果测得m PEG_p ALD主成分的重均分子量(Mw)为19 444,分布指数(D)为1.01;杂质1的Mw为38 703,D为1.01,含量为1.31%;杂质2的Mw为61 036,D为1.00,含量为0.70%。与常规凝胶渗透色谱(GPC)相比,该方法分辨率高,分析速度快,能快速测定m PEG_p ALD的相对分子量及其分布,并能得到其纯度和杂质含量,为其工艺研发、质量控制提供了科学的依据,同时也可用于其它PEG衍生物的相对分子量及其分布和纯度的测定。     

Determination of the enantiomeric purity of levodopa, methyldopa, carbidopa and tryptophan by use of chiral mobile phase high-performance liquid chromatography

Chiral mobile phase high-performance liquid chromatography was used successfully for the determination of the enantiomeric purity of levodopa, methyldopa, carbidopa and tryptophan. The method investigated uses phenylalanine and copper sulfate in the mobile phase and a C18 column. Linearity, precision, accuracy, detection limit and interference from expected impurities were assessed. The method is also applicable to the measurement of enantiomeric purity in levodopa tablets and capsules.     

Reversed phase ion-pair high performance liquid chromatographic gradient separation of related impurities in 2,4-disulfonic acid benzaldehyde di-sodium salt

A reversed phase ion-pair gradient liquid chromatographic method has been developed and validated for purity determination of the hydrophilic compound 2,4-disulfonic acid benzaldehyde di-sodium salt (2,4-DSAD) containing both hydrophilic and more lipophilic related impurities. Mixtures of acetonitrile-phosphate buffer containing tetrahexylammonium hydrogen sulfate as the ion-pairing reagent were used as the mobile phase. A linear gradient, which generated simultaneous change in the concentration of organic modifier, buffer concentration and the concentration of ion-pairing reagent, was applied. The method allows detection of impurities at low levels (0.01% w/w). Excellent repeatability for both retention time (RSD< or =0.3%, n = 6) and detector response (RSD = 0.03%, n = 6 for the main peak and RSD = 6%, n = 6, for an impurity at 0.01 area% level) was obtained. The method was shown to be robust for routine analysis and has been successfully transferred to the quality control laboratory.     

Ion-Pair RP-LC of Tegaserod Maleate and its Impurities in Pharmaceutical Formulations and in Dissolution Studies

A simple ion-pair reversed-phase high-performance liquid chromatographic (RP-HPLC) method has been developed for determination of tegaserod maleate and related impurities in tablet dosage forms. The mobile phase was 60:40 (v/v) acetonitrile-25 mmol L^?1 sodium dodecyl sulfate, adjusted to pH 2.6 with glacial acetic acid. A C18 column was used as stationary phase and UV detection was at 314 nm. The method was optimized and validated. Response was linearly dependent on concentration between 0.1 and 100 µg mL^?1 with a limit of quantification (LOQ) of 0.1 µg mL^?1 for tegaserod maleate (S/N = 10). Under optimum conditions, tegaserod maleate was successfully separated from related substances, including 5-methoxyindole-3-carboxaldehyde remaining after synthesis and other impurities possibly resulting from oxidization and decomposition. The excipients did not interfere with assay of tegaserod maleate in tablet dosage forms. It is suggested that the proposed method can be used for routine quality control and dosage-form assay of tegaserod maleate.     

Analysis of Spectinomycin by HPLC with Evaporative Light-Scattering Detection

A high-performance liquid chromatographic method with evaporative light-scattering detection (ELSD) has been developed for analysis of spectinomycin and related impurities. Separation of spectinomycin from structurally related impurities was achieved on a C₁₈ column. The optimized mobile phase was 25 mmol L⁻¹ ammonium acetate (pH 7.5)-methanol, 90:10 (v/v), at a flow rate of 0.6 mL min⁻¹. The temperature of the drift tube of the ELSD was 95°C and the flow rate of carrier gas was 2.2 L min⁻¹. The accuracy, specificity, precision, linearity, sensitivity, and robustness of the method were validated in accordance with ICH guidelines. In addition to determination of spectinomycin and related impurities, the method is also ideal for determination of the salts spectinomycin hydrochloride and spectinomycin sulfate.     

Development of an ion-pair reversed-phase HPLC method with indirect UV detection for determination of phosphates and phosphites as impurities in sodium risedronate

A method based on RP-HPLC with indirect UV detection was developed for the determination of phosphates and phosphites as impurities in sodium risedronate. RP separation of the phosphates and phosphites was achieved by adding tetrabutylammonium hydroxide as an ion-pairing agent in the mobile phase. Potassium hydrogen phthalate was added to the mobile phase as an ionic chromophore in order to obtain high background absorption of the mobile phase. Separation was performed on a C18 column using a mixture of pH 8.2 buffer (containing 0.5 mM tetrabutylammonium hydroxide and 1 mM phthalate) and acetonitrile (95 + 5, v/v) as the mobile phase, with indirect UV detection at 248 nm. The validation of the method included determination of specificity/selectivity, linearity, LOD, LOQ, accuracy, precision, and robustness. The LOD was 0.86 microg/mL for phosphates and 0.76 microg/mL for phosphites. The LOQ was 2.60 microg/mL for phosphates and 2.29 microg/mL for phosphites. The developed method is suitable for quantitative determination of phosphates and phosphites as impurities in QC of sodium risedronate.     

利尿剂的胶束色谱分析

用十二烷基硫酸钠（SDS）胶束溶液作为流动相，建立了11种利尿剂的胶束色谱系统分析方法；研究了SDS浓度、流动相的pH、有机改性剂及温度对色谱保留行为的影响，优化了色谱条件，并且不经化学处理直接进样测定了人尿中的烯睾丙内酯；该法适用于兴奋剂中利尿剂的常规分析。     

Selective determination of famotidine in human plasma by high performance liquid chromatography in alkaline media with solid phase extraction

A new method is described for the determination of famotidine by solid phase extraction from alkalinized human plasma followed by reversed phase (RP) HPLC in acetonitrile/alkaline buffer with molsidomine as an internal standard. Different acetonitrile/aqueous buffer mobile phases as well as various RP columns were used. Alkaline medium allowed the limit of quantitation to be lowered to 5 ng/mL of plasma as the famotidine gives more intense absorption at about 286 nm (at pH values higher than 7). Moreover, work in alkaline media and at this wavelength is highly selective as peaks corresponding to impurities present in most samples are well separated. A method using a mildly alkaline mobile phase (acetonitrile/10 mM phosphate with 10 mM 1‐heptanesulphonic acid, pH 7.5) was successfully used for determination of famotidine in human plasma in a pharmacokinetic study.     

Development and Validation of Rapid Resolution RP-HPLC Method for Simultaneous Determination of Atorvastatin and Related Compounds by Use of Chemometrics

Abstract

A Rapid Resolution Reversed Phase High-Performance Liquid Chromatography (RR RP-HPLC) method has been developed and validated for the simultaneous determination of atorvastatin and seven related compounds specified as impurities. Experimental design was used during method optimization (full factorial 3² design) and robustness testing (central composite design). Chromatography was performed with mobile phase containing phosphate buffer pH 3.5 and a mixture of 10% (v/v) tetrahydrofuran in acetonitrile as organic modifier. A Zorbax Eclipse XDB C18 Rapid Resolution HT 4.6 mm × 50 mm, 1.8 µm particle size column was used. The developed method allowed determination of Atorvastatin Calcium (ATV Ca) purity and level of impurities in drug substances.     

A validated rapid stability-indicating method for the determination of related substances in solifenacin succinate by ultra-fast liquid chromatography

A rapid, sensitive, and accurate ultra-fast liquid chromatographic method is developed for the determination of related substances and degradants of Solifenacin Succinate, an active pharmaceutical ingredient used in the treatment of overactive bladder. Chromatographic separation of Solifenacin Succinate, its related substances, and degradants was achieved using a Shimpack XR-ODS-II column and mobile phase system containing 10 mM potassium dihydrogen orthophosphate in water. The pH of the buffer was adjusted to 7.0 using triethyl amine (mobile phase A). LC-grade acetonitrile was used as mobile phase B, employing a binary-gradient program at a flow rate 0.5 mL/min. The resolution between the critical pair of peaks (Impurity A and analyte) was found to be greater than 3.5. The limits of detection and quantification (LOQ) of Impurity A, Impurity B, and the analyte were 0.2 and 0.6 μg/mL, respectively for a 5-μL injection volume. The percentage recovery of impurities in the presence of sample matrix ranged from 95 to 104 w/w. The test solution and mobile phase was observed to be stable up to 24 h after the preparation. The validated method yielded good results of precision, linearity, accuracy, robustness, and ruggedness. The proposed method is found to be rapid, accurate, and suitable for the quantitative determination of related substances and degradants during quality control of Solifenacin Succinate active pharmaceutical ingredient.     

Reversed‐Phase High‐Performance Liquid Chromatographic Separation and Determination of 4‐Amino‐azobenzene‐4′,5‐disulfonic Acid and Related Products in Industrial Wastewater Effluents

A simple and rapid reversed‐phase high‐performance liquid chromatographic method for the separation and determination of 4‐amino‐azobenzene‐4′,5‐disulfonic acid (AABDS) and its process‐related impurities was developed. The separation was achieved on a μ‐Bondapak C₁₈ column using 0.15 M ammonium sulfate‐acetonitrile (55:45) (v/v) as eluent. A UV‐visible spectrophotometric detector fixed at 386 nm was used both for detection and quantitation. The method was used not only for quality assurance but also for process development and wastewater management of AABDS.