注射用头孢西丁钠杂质谱分析方法影响因素探讨

目的 采用计算机辅助设计试验,考察影响注射用头孢西丁钠有关物质色谱分析方法的主要因素并使目标因子达到最优,对注射用头孢西丁钠杂质谱分析方法进行优化。方法 制备系统适用性混合溶液,确定目标因子。通过Plackett-Burman(PB)设计筛选显著因素;通过中心组合设计(CCD)设计进行优化并确定最佳色谱条件;采用质谱法分析有关物质结构。结果 色谱柱参数、流动相参数和梯度程序等因素均对杂质分离分析有较大影响;在建立的最优色谱条件下,11个潜在杂质均能得到有效分离并被检测。杂质谱分析显示,不同企业产品杂质谱存在差异。结论 使用计算机辅助设计优化色谱分析方法,可实现对潜在杂质的有效分析,为完善注射用头孢西丁钠质量标准和质量控制提供了指导。     

青霉素发酵过程优化控制问题及方法研究

对各种过程优化控制方法的特点及其在青霉素发酵过程优化控制中的应用情况进行了总结,指出了目前青霉素发酵过程建模和优化控制中存在的主要问题,并在对青霉素发酵过程复杂性进行分析的基础上提出了解决方法。     

青霉素类抗生素的聚合物分析

摘要：在青霉素类药物杂质谱的控制中,对聚合物类杂质的分析是最薄弱环节,本文对近年来青霉素类抗生素在聚合物分析中取得的进展进行了综述。凝胶色谱-柱切换-质谱分析技术的广泛应用,使得人们对青霉素的聚合反应机理、结构等的认知不断完善,已逐渐形成了相对系统的聚合物分析策略与方法。通过理论计算结合强制聚合反应是进行聚合物分析的较理想方案;利用LC-MS在线分析技术,通过选择适宜的质谱条件获取聚合物聚合位点的结构信息,是推测聚合物结构的最有效途径;通过对各类聚合物质谱裂解规律的系统总结,有助于对聚合物结构的解析。     

超高效液相色谱与高效液相色谱方法转换及验证北大核心CSCD

超高效液相色谱(UPLC)分离原理与高效液相色谱(HPLC)完全一致,但分离度更好,灵敏度更高,并且速度快,消耗溶剂少。如果实现两者方法的转换,并进行必要的验证工作,可在不变更标准的前提下合规应用UPLC技术,对药品质量进行高效精准的控制。中国药典2015年版四部关于UPLC的应用有简单说明,但是涉及与HPLC方法的转换和验证没有细节规定,使该技术推广应用受阻。本文从2种仪器的差异进行分析,阐述了方法转换时如何进行参数变更,包括色谱柱、流速、进样体积和梯度洗脱程序的转换。基于转换前方法已经经过全面验证,转换后的方法验证应主要着眼于仪器带来的差异,需验证分离度和灵敏度2个方面。本文根据转换前方法的不同情况,对方法验证细节要求进行了分析讨论,以确保方法转换后准确有效,合乎规范。     

生物合成调控的青霉素发酵数学模型与过程优化

青霉素生物合成受溶解氧、溶解二氧化碳、pH、氨氮、碳源(特别是葡萄糖)等的调控，这些调控反应的产生不仅与基础培养基配方有关，更受发酵过程通气、搅拌条件及补料方案的影响。为此，笔者通过把握发酵过程中产生菌生长和青霉素生物合成代谢流的元素平衡、能量平衡以及传递与反应速度平衡的方法，结合生产经验和数据资料，建立了一种能够模拟青霉素发酵过程工艺学参数和经济学参数变化的数学模型。应用这一模型，在充分考虑生物合成代谢调控的基础上，对青霉素发酵过程进行优化，即通过补水维持上述平衡，避免因环境条件、初始条件和约束条件的变化及人为的失误造成的过程波动，使生产不断趋向最优状态。模拟运行表明，这种优化可显著提高发酵生产的经济效益。     

青霉素V钾及其有关物质分析方法的比较

目的：中国药典2005年版（ChP 2005）中青霉素V钾有关物质分析方法与其他药典及文献报道方法在流动相、检测波长、杂质限度等方面存在较大差异，本文比较了几种典型的青霉素V钾有关物质分析方法，为完善中国药典中青霉素V钾有关物质控制方法提供科学依据。方法：采用SinoChrom ODS—BP柱（150mm×4．6mm，5μm），柱温35℃，以青霉素V钾及其杂质的分离情况为指标，比较了ChP 2005方法和文献报道方法[流动相组成同欧洲药典5．1版方法，但采用等度洗脱]的专属性；由于在pH3．5时，醋酸盐缓冲系统的缓冲能力优于磷酸盐缓冲系统，故考察了将文献报道方法中pH3．5磷酸盐缓冲液换为pH3．5醋酸盐缓冲液后系统的专属性变化；比较了青霉素V钾及其有关物质的UV图谱，为检测波长的合理选择提供依据。结果：文献报道方法的专属性优于ChP 2005方法，能将青霉素V钾及其有关物质较好地分离；将pH3．5磷酸盐缓冲液换为pH3．5醋酸盐缓冲液后，方法的专属性无明显改变；除青霉素外，青霉素V及其杂质的uV吸收值均为：A（254nm）〈A（268nm）〈A（220nm），但考虑到220nm接近UV末端检测，容易受到试剂等因素的影响，故建议青霉素V钾有关物质控制采用268nm检测。结论：基于本文结果，为完善ChP 2005中青霉素V钾质量标准，建议：（1）将青霉素V钾原料及制剂中有关物质检查方法中的流动相改为专属性更好的文献方法流动相或者改进后的流动相，检测波长仍采用原方法的268nm检测；（2）为促进青霉素V钾原料及制剂产品质量的提高，建议有关物质检查项中增加单一杂质的限度，采用外标法或者校正因子法控制单一杂质。     

国产注射用阿奇霉素杂质谱及其与成盐工艺相关性的考察

目的:考察国产不同成盐注射用阿奇霉素所含杂质谱情况,并对其工艺进行相关性评价。方法:以217批注射用阿奇霉素有关物质HPLC检验数据为基础,采用数理统计与化学计量学方法,对其杂质谱及与成盐工艺相关性进行考察,并通过加速实验验证考察结果。结果:所检样品中均检出阿奇霉素B(EP-B)和3′-去克拉定糖阿奇霉素(EP-J)杂质,且在不同厂家、不同成盐工艺产品中变化幅度较大,统计标准偏差(SD)分别达2.5和2.0;其他杂质还检出阿奇霉素-N-氧化物(EP-L)、氮红霉素(EP-A)。结论:目前国产注射用阿奇霉素中主要杂质为阿奇霉素-N-氧化物、3′-去克拉定糖阿奇霉素、氮红霉素和阿奇霉素B;氮红霉素和阿奇霉素B为工艺杂质,分别源于合成反应残留和起始原料红霉素中红霉素B残留;阿奇霉素-N-氧化物和3′-去克拉定糖阿奇霉素分别为阿奇霉素氧化和酸降解物,其中酸降解物为主要降解形式;不同厂家注射用阿奇霉素中所含杂质的种类、数目及水平均存在较大差异,且与其成盐工艺密切相关。     

国产罗红霉素杂质谱及与合成工艺相关性评价

目的:考察国产罗红霉素的杂质谱情况,并与合成工艺相关性进行初步评价。方法:采用欧洲药典7.0版罗红霉素有关物质方法对国产样品进行分析,使用Waters Symmetry ShieldTMC18色谱柱(150 mm×3.9 mm,5μm),以0.52 mol·L-1磷酸二氢铵溶液[用氢氧化钠溶液(8.5→100)调节p H至4.3]-乙腈(74∶26)为流动相A,以水-乙腈(30∶70)为流动相B,梯度洗脱(0~50 min,100%A;50~51 min,100%A→90%A;51~80 min,90%A;80~81 min,90%A→100%A;81~100 min,100%A),色谱柱温15℃,自动进样器温度8℃,检测波长205 nm。以系统适用性对照品对11种已知杂质进行定位,以罗红霉素为外标计算杂质含量。结果:国产罗红霉素中检出的主要杂质为杂质H、杂质F和杂质C。杂质H是影响国产罗红霉素质量的主要杂质。结论:国产罗红霉素的主要杂质H和杂质F与合成工艺中起始反应物硫氰酸红霉素中红霉素B和去甲基红霉素的含量相关;杂质C为反应中间体红霉素(E)肟的残留,通过优化纯化工艺可降低杂质C的含量。国产罗红霉素的合成工艺可有效的去除侧链合成中引入的杂质G、杂质J和杂质K。     

国产盐酸多西环素原料中有关物质的结构鉴定及杂质谱考察

目的:鉴定盐酸多西环素原料中检出的有关物质结构,考察国产原料的杂质谱.方法:用十八烷基硅烷键合硅胶为填充剂(pH值适用范围应大于9);以醋酸盐缓冲液[0.25mol·L-1醋酸铵-0.1 mol·L-1乙二胺四醋酸二钠-三乙胺(100:10:1),用冰醋酸或氨水调节pH值至8.8]-乙腈(85:15)为流动相;柱温35...     

头孢尼西钠杂质谱与市售品杂质谱的相似度研究

目的采用反相高效液相色谱法研究自制头孢尼西钠(CFNS)杂质谱和市售CFNS杂质谱的相似性。方法采用自身对照法对杂质谱进行定量测定,用相似度法比较自制CFNS杂质谱和市售CFNS杂质谱的相似性,并用UPLC-MS初步鉴定杂质种类。结果自制CFNS和市售CFNS的杂质谱均为9个共有杂质,两者杂质谱相似度均>0.99,自制CFNS杂质总含量略低于市售CFNS。结论自制CFNS杂质谱中杂质种类和总含量与市售CFNS杂质谱中杂质种类和总含量相当,中药指纹图谱相似度评价方法完全适合评价CFNS的杂质谱。     

Implementation of two sustainable chromatographic methods for the simultaneous micro-quantitation and impurity profiling of metformin and rosuvastatin in recently approved fixed dose pills: Greenness and whiteness studies

Rosuvastatin-Metformin is an emerging fixed dose combination (FDC) for the treatment of diabetes mellitus-associated dyslipidemia. Generally, the beneficial outcomes of FDC in reduction of pills burden, cost and complexity of medications protocols, encouraged analysts and we in turn for the development of rapid sustainable analytical methods for the simultaneous estimation of such important drug combinations. Within this framework, two validated analytical methodologies have been implemented for the neat separation and quantitation of metformin (MET) and rosuvastatin (RSV) in presence of the pharmacopeial impurities cyanoguanidine (CYG) (impurity A of MET) and the rosuvastatin-related substance B (RSV-impurity B). The latter was freshly prepared and then confirmed using Liquid Chromatography-Mass spectrometry (LC-MS). Method I represented the first electro-driven separation method for the assay of this FDC using Micellar Electrokinetic Chromatography (MEKC). The optimized conditions were 50 mM borate buffer containing 100 mM sodium dodecyl sulfate (SDS) at pH 9.2 using a deactivated fused silica capillary (50 cm effective length × 50 μm id). Method II was High Performance Liquid Chromatography (HPLC) method using Microsorb MV-C18 (4.6 × 250 mm, 5 μm particle size) column with 20 mM sodium dihydrogen phosphate buffer (pH 3.5), acetonitrile and methanol as mobile phase constituents. The separation was achieved with a linear gradient program and flow rate 1.5 mL/min all over the run. The Diode Array Detector (DAD) detector was set at wavelengths 243 nm for RSV and 235 nm for MET in both methods. The 4 peaks were separated in about 8 and 5 min for methods I and II, respectively. Linear relationships were obtained between peak areas and concentrations of metformin and rosuvastatin in the ranges 10–100 μg/mL for MEKC and 1–150 μg/mL for HPLC. The 2 techniques fulfilled the International Council for Harmonization (ICH) validation criteria, as well as the green and white analytical chemistry principles which were evaluated using the trendiest AGREE, hexagon and RGB12 tools. The interpretation of scores indicated that the suggested methods outperformed the reported methods in terms of eco-friendliness and sustainability. They offered fast, feasible, stability indicating and accurate analysis of both drugs in bulk and in pills.     

Estimation of impurity profiles of drugs and related materials: part XXI. HPLC/UV/MS study of the impurity profile of ethynodiol diacetate

The impurity profile of ethynodiol diacetate was investigated using the HPLC/UV/MS method. Using the slightly modified HPLC method of USP 24 two impurities, earlier isolated by preparative HPLC and investigated by NMR spectroscopy were separated and characterised. The mass spectra amended by the diode-array UV spectra supported the earlier found structures (E and Z isomers of 17-ethinyl-estr-4-ene-3β,17-diol-3-acetate-17-(3′-acetoxy-2′-butenoate). Another, hitherto not described impurity, 17-ethinyl-estr-4-ene-3β,17-diol-3-acetate-17-(3-oxo-butanoate) has also been separated and characterised by means of its mass spectrum, NMR and UV spectra.     

左乙拉西坦中有关物质检查方法的比较研究

目的 比较并讨论《美国药典》35版、《欧洲药典》8.0版、《英国药典》2013年版和文献中左乙拉西坦有关物质的检测方法,为左乙拉西坦中有关物质检测方法的选择提供依据.方法 采用3种方法分析左乙拉西坦的已知杂质、强制降解样品、左乙拉西坦及其注射液,比较各方法的专属性、灵敏度.结果 采用《美国药典》的方法检出5个已知杂质,未知杂质数目较多,专属性良好;采用《欧洲药典》的方法检出2个已知杂质,未知杂质数目少;采用文献方法检出5个已知杂质,未知杂质数目较少,杂质间分离度不好.6批样品中,采用《美国药典》方法和《欧洲药典》方法检出的总杂质量相近,文献方法中检出的总杂质量略少.结论 《美国药典》的方法最适宜左乙拉西坦中有关物质的检查.     

Estimation of impurity profiles in drugs and related materials. Part 11 — the role of chromatographic and spectroscopic methods in the estimation of side-reactions in drug syntheses

Impurities in drugs are classified on the basis of the types of side-reactions in drug syntheses resulting in their formation. This is shown by summarizing the authors' earlier results in the field of impurity profiling of 19-nor-steroids, ethynodiol diacetate, mazipredone, pipecuronium bromide, flumecinol, enalapril, pyridinol carbamate, phenylbutazone, thymotrinan and some new results related to danazol and famotidine.     

Estimation of impurity profiles of drugs and related materials. Part VIII: Combined application of high- performance liquid chromatography and NMR spectroscopy in the impurity profiling of drugs

The usefulness of the joint application of HPLC and NMR spectroscopy in drug impurity profiling is demonstrated by the following examples: (1) identification of Z and E isomers of 17 alpha-ethynyl-4-oestrene-3 beta, 17-diol-3-acetate-17-(3'-acetoxy-2'-butenoate) in ethynodiol diacetate; (2) identification of the p-tolyl analogue as the impurity of enalapril maleate; (3) identification and quantification of 2'-dehydro-pipecuronium bromide in pipecuronium bromide. The possibilities of utilizing NMR spectroscopy for the identification and quantification of the impurities with and without their isolation are discussed.     

Design space approach in the optimization of the spray-drying process

From a quality by design perspective, the aim of the present study was to demonstrate the applicability of a Bayesian statistical methodology to identify the Design Space (DS) of a spray-drying process. Following the ICH Q8 guideline, the DS is defined as the “multidimensional combination and interaction of input variables (e.g., materials attributes) and process parameters that have been demonstrated to provide assurance of quality.” Thus, a predictive risk-based approach was set up in order to account for the uncertainties and correlations found in the process and in the derived critical quality attributes such as the yield, the moisture content, the inhalable fraction of powder, the compressibility index, and the Hausner ratio. This allowed quantifying the guarantees and the risks to observe whether the process shall run according to specifications. These specifications describe satisfactory quality outputs and were defined a priori given safety, efficiency, and economical reasons. Within the identified DS, validation of the optimal condition was effectuated. The optimized process was shown to perform as expected, providing a product for which the quality is built in by the design and controlled setup of the equipment, regarding identified critical process parameters: the inlet temperature, the feed rate, and the spray flow rate.     

Quality by design approach in the optimization of the spray-drying process

Context/objective: The aim of this study was to illustrate the influence of the processing parameters, inlet temperature, atomization air flow rate and feed flow rate, on critical quality attributes of spray-dried powders using design of experiments (DoE).

Methods: Spray-dried powders were characterized by laser diffraction, X-ray powder diffraction (XRPD) and near-infrared spectroscopy (NIR). Multivariate analysis of two different experimental designs was performed to elucidate the optimal process conditions.

Results and discussion: XRPD revealed that the spray-dried powders consisted of crystalline β-mannitol and amorphous trehalose. Non-invasive NIR measurement was successfully used for correlating the critical quality attribute particle size with size determined by laser diffraction. The full factorial design proved to be unsuitable due to the non-linear influence of factors. The composite face-centered design improved the quality of the models and showed both linear and non-linear influence of the parameters on the outcomes. A model explaining the influence of the factors on all quality attributes showed similar results as the models optimized for a single response.

Conclusion: This study showed the applicability of DoE for the investigation of spray-dried powders. The knowledge of the interplay between process parameters and quality attributes will enable rational process design to achieve a desired outcome.     

Development and application of a validated HPLC method for the determination of gabapentin and its major degradation impurity in drug products

A simple isocratic reversed-phase HPLC method for the determination of gabapentin and its major degradation impurity, 3,3-pentamethylene-4-butyrolactam, was developed and validated for use in the analysis of pharmaceutical tablets and capsules. Separation was achieved on a Brownlee Spheri-5 Cyano column using an acetonitrile–10 mM KH₂PO₄/10 mM K₂HPO₄ (pH 6.2) (8:92, v/v) mobile phase. The compounds were eluted isocratically at a flow rate of 1 mL/min. Both compounds were analyzed with UV detection at 210 nm. The method was validated according to USP Category I requirements for gabapentin and USP Category II for 3,3-pentamethylene-4-butyrolactam. The validation characteristics included accuracy, precision, linearity, range, specificity, limit of quantitation and robustness. Validation acceptance criteria were met in all cases. This method was used successfully for the quality assessment of four gabapentin drug products.     

Development and optimization of modified nucleosides and deoxynucleosides simultaneous extraction with the use of Design of Experiments approach

Modified nucleosides and deoxynucleosides are investigated as potential markers of pathological disorders. Due to different molecular properties, their concurrent extraction from biological matrices is challenging. The aim of the present work was the development of sample preparation procedure allowing for simultaneous extraction of 12 selected modified nucleosides and deoxynucleosides with the use of solid phase extraction (SPE) technique. Commercially available mixed-mode SPE cartridges based on phenylboronic acid and cation-exchange sorbents were utilized. In order to decrease the number of experiments and, consequently the volume of applied solvents, the developed method was optimized with the use of Design of Experiments approach. Optimization of SPE method was carried out by employing Fractional Factorial Design with selected input variables such as composition of conditioning, washing and elution solvents. Next, Full Factorial Design with conditioning, extraction and elution duration time as factors was employed. Evaluated responses in these two experimental sets were peak areas of selected analytes. Levels of assessed factors were determined. Developed and optimized SPE procedure allowed for simultaneous extraction of modified nucleosides and deoxynucleosides with mean recovery of 76.0%. Procedure was applied on the exemplary urine samples with the use of high performance liquid chromatography hyphenated with triple quadrupole mass spectrometry (HPLC-QqQ/MS).