Preparation Process and Thin Layer Chromatography Identification of Wufang Babu Poultice

[Objectives] To establish the process flow of preparation of Wufang Babu Poultice and the identification method of thin layer chromatography (TLC). [Methods] For the forming process of Wufang Babu Poultice, the preparation method of mixed pharmaceutical powder with suitable pharmaceutical excipients was adopted. Qualitative identification of medicinal materials in Wufang Babu Poultice (Strychni Semen, Rhei Radix Et Rhizoma, Dipsaci Radix, and Angelicae Sinensis Radix) was carried out by TLC. [Results] Mixed pharmaceutical powder mixed with glycerol, gelatin and other pharmaceutical excipients can be prepared for forming. The test solution chromatography of each medicinal material (Strychni Semen, Rhei Radix et Rhizoma, Dipsaci Radix, Angelicae Sinensis Radix) showed pigment spots of the same color at the same position as its corresponding control medicinal materials and reference chromatography, and the display was clear. [Conclusions] The preparation process is simple and feasible, and can be used as the forming process of Wufang Babu Poultice. The TLC determination method is simple to operate, has good specificity, and has no effect on negative results, and can be used for identification of Wufang Babu Poultice.

Isolation and Purification of Unstable Iridoid Glucosides from Traditional Chinese Medicine by Preparative High Performance Liquid Chromatography Coupled with Solid-phase Extraction

An efficient preparative method was successfully developed for isolation and purification of unstable components from medicinal plant extracts, using a combined method of preparative high performance liquid chro-matography(HPLC) and solid-phase extraction(SPE). The aim of this study was to obtain an effective method with high preparative efficiency and importantly to avoid the transformation of unstable compounds. The preparative HPLC system was based on an LC/MS controlled four-channel autopurification system. The SPE method was performed with a C18 packing material to trap the target compounds and to remove the acidic additive derived from the mobile phase. Using this method, the unstable iridoid glucosides(IGs) as model compounds were successfully isolated and purified from the extract of Hedyotis diffusa Willd. Six IGs(including one new minor IG) and one nucleotide compound were simultaneously obtained, each with a purity of 91% as determined by HPLC. The structures of the isolated compounds were identified by UPLC/Q-TOF MS, UV, 1D and/or 2D NMR. It was demonstrated that the combination of preparative HPLC with SPE is a versatile tool for preparative purification of unstable compounds from complex natural products.

Development of Preparative Chromatography for Proteomic Approach of Mycorrhizal Symbiosis

Although mechanism of symbiosis between arbuscular mycorrhizal fungi (AMF) and host plants has been investigated by genetic analysis, very little knowledge has been obtained because genome analysis of AMF is not perfect yet. Thus, we tried to develop mass purification of proteins using preparative chromatography in order to accelerate roteomic analysis of proteins related to mycorrhizal symbiosis, such as 24 and 53 kDa proteins. In particular, our data showed that 53 kDa proteins would be restrictively expressed when mycorrhizal fungi and host plants were stressed. However, 24 kDa proteins, which appear to be a usable indicator for the existence of various my-corrhizal fungi, were habitually detected in not only AMF but also other mycorrhizal fungi such as ectomycorrhizal fungi (EF). Moreover, we discovered new preparative chromatographical techniques for isolation and mass purification of those proteins. We are convinced that this chromato-graphical technique will greatly contribute to proteomic approach of mycorrhizal symbiosis.

PREPARATIVE STUDIES ON MONODISPERSED POLYMERIC NICROPARTICLES AND LATEX-COATED PACKINGS IN ANION CHROMATOGRAPHY

Monodispersed potymeric microparticles were prepared by seed-poly- merization.High performance packings were obtained for anion chromatography by coating the surface-sutfonated partictes with quarternized latexes.

Advancements in the preparation of high-performance liquid chromatographic organic polymer monoliths for the separation of small-molecule drugs

The various advantages of organic polymer monoliths, including relatively simple preparation processes,abundant monomer availability, and a wide application range of pH, have attracted the attention of chromatographers. Organic polymer monoliths prepared by traditional methods only have macropores and mesopores, and micropores of less than 50 nm are not commonly available. These typical monoliths are suitable for the separation of biological macromolecules such as proteins and nucleic acids, but their ability to separate small molecular compounds is poor. In recent years, researchers have successfully modified polymer monoliths to achieve uniform compact pore structures. In particular, microporous materials with pores of 50 nm or less that can provide a large enough surface area are the key to the separation of small molecules. In this review, preparation methods of polymer monoliths for high-performance liquid chromatography, including ultra-high cross-linking technology, post-surface modification, and the addition of nanomaterials, are discussed. Modified monolithic columns have been used successfully to separate small molecules with obvious improvements in column efficiency.

Quantification of six bioactive compounds in Zhenqi Fuzheng preparation by high-performance liquid chromatography coupled with diode array detector and evaporative light scattering detector

A simple and accurate high-performance liquid chromatography（HPLC）coupled with diode array detector（DAD）and evaporative light scattering detector（ELSD）was established for the determination of six bioactive compounds in Zhenqi Fuzheng preparation（ZFP）.The monitoring wavelengths were 254,275 and 328 nm.Under the optimum conditions,good separation was achieved,and the assay was fully validated in respect of precision,repeatability and accuracy.The proposed method was successfully applied to quantify the six ingredients in 31 batches of ZFP samples and evaluate the variation by hierarchical cluster analysis（HCA）,which demonstrated significant variations on the content of these compounds in the samples from different manufacturers with different preparation procedures.The developed HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of this preparation.

Studies on the Renaturation with Simultaneous Purification of Recombinant Human Proinsulin with Unit of Simultaneous Renaturation and Purification of Protein in Semi-preparative Scale

The renaturation and purification of recombinant human proinsulin (rh-proinsulin) expressed in E. coli with the unit of simultaneous renaturation and purification of protein (USRPP) in semi-preparative scale was studied. The result shows that rh-proinsulin extracted with 8.0 mol/L urea can be renatured and purified simultaneously in 45 minutes with the USRPP (1050 mm ID). The purity of rh-proinsulin was found to be more than 90% and the mass recovery to be more than 80%. The renaturation effect of rh-proinsulin with the USRPP was tested by enzyme cleavage for obtaining insulin. In addition, the result was further confirmed with RPLC, SDS-PAGE electrophoresis, and MALDI-TOF, respectively.

Determination of Camylofin Dihydrochloride and Nimesulide in Pharmaceutical Preparation by Gas Chromatography

This research paper describes simple analytical method for determination of Camylofin dihydrochloride and Nimesulide in tablet formulation by Gas chromatography method. Benzoic acid was used as internal standard. Validation was carried out in compliance with the International Conference on Harmonization guidelines. The method utilized GC (Agilent Technologies 6890 N Network GC system with FID detector), and RTX-5 capillary column (5% diphenyl-95% dimethyl polysiloxane), 30 m × 0.53 mm, 1.5 μm as stationary phase. Helium was used as the carrier gas at a flow rate of 1.5 mL?min–1. The proposed method was validated for linearity, LOD, LOQ, accuracy, precision, ruggedness and solution stability. It can be conveniently adopted for routine quality control analysis.

Determination of diclofenac in pharmaceutical preparations by voltammetry and gas chromatography methods

Rapid, sensitive and specific methods were developed for the determination of diclofenac in pharmaceutical preparations by linear sweep voltammetry （LSV） and gas chromatography （GC） with mass spectrometry （MS） detection. The linearity was established over the concentration range of 5- 35 μg/mL for LSV and 0.25-5 μg/mL for GC-MS method. The intra- and inter-day relative standard deviation （RSD） was less than 4.39% and 4.62% for LSV and GC-MS, respectively. Limits of quantification （LOQ） were determined as 4.8 and 0.15 μg/mL for LSV and GC-MS, respectively. No interference was found from tablet excipients at the selected assay conditions. The methods were applied for the quality control of commercial diclofenac dosage forms to quantify the drug and to check the formulation content uniformity.

Adsorbents for Expanded Bed Adsorption： Preparation and Functionalization

Expanded bed adsorption(EBA),a promising and practical separation technique,has been widely studied in the past two decades.The development of adsorbents for EBA process is a challenging course,with the special design and preparation according to the target molecules and specific expanded bed systems.Many types of supporting matrices for expanded bed adsorbents have been developed,and their preparation methods are being consummated gradually.These matrices are activated and then coupled with ligands to form functionalized adsorbents,including ion-exchange adsorbents,affinity adsorbents,mixed mode adsorbents,hydrophobic charge induction chromatography adsorbents and others.In this review,the preparation of the matrices for EBA process is summa-rized,and the coupling of ligands to the matrices to prepare functionalized adsorbents is discussed as well.

Pharmacokinetics and bioequivalence of ranitidine and bismuth derived from two compound preparations

AIM： To evaluate the bioequivalence of ranitidine and bismuth derived from two compound preparations. METHODS： The bioavailability was measured in 20 healthy male Chinese volunteers following a single oral dose （equivalent to 200 mg of ranitidine and 220 mg of bismuth） of the test or reference products in the fasting state. Then blood samples were collected for 24 h. Plasma concentrations of ranitidine and bismuth were analyzed by high-performance liquid chromatography and inductively coupled plasma-mass spectrometry （ICPMS）, respectively. The non-compartmental method was used for pharmacokinetic analysis. Log-transformed Cmax, AUC（0-t） and AUC（0-∞） were tested for bioequivalence using ANOVA and Schuirmann two-one sided t-test. Tmax was analyzed by Wilcoxon＇s test. RESULTS： Various pharmacokinetic parameters of ranitidine derived from the two compound preparations, including Cmax, AUC（0-t）, AUC（0-∞）, Tmax and T1/2, were nearly consistent with previous observations. These parameters derived from test and reference drug were as follows： Cmax（0.67 ± 0.21 vs 0.68 ± 0.22 mg/L）, AUC（0-t）（3.1 ± 0.6 vs 3.0 ± 0.7 mg/L per hour）, AUC（0-∞）（3.3 ± 0.6 vs 3.2 ± 0.8 mg/L per hour）, Tmax （2.3 ± 0.9 VS 2.1 ± 0.9 h） and T1/2 （2.8 ± 0.3 vs 3.1 ± 0.4 h）. In addition, double-peak absorption profiles of ranitidine were found in some Chinese volunteers. For bismuth, those parameters derived from test and reference drug were as follows： Cmax （11.80 ± 7.36 vs 11.40 ± 6.55 μg/L）, AUC（0-t） （46.65 ± 16.97 vs 47.03 ± 21.49 μg/L per hour）, Tmax （0.50 ± 0.20 vs 0.50 ± 0.20 h） and T1/2 （10.2 ± 2.3 vs 13.0 ± 6.9 h）. Ninety percent of confidence intervals for the test/reference ratio of Cmax, AUC（0-t） and AUC（0-∞） derived from both ranitidine and bismuth were found within the bioequivalence acceptable range of 80%-125%. No significant difference was found in Tmax derived from both ranitidine and bismuth. CONCLUSION： The two compound preparations are bioequivalent and may be prescribed interchangeably.

全自动固相萃取—高效液相色谱法测定现制饮料中7种合成着色剂

目的:建立一种全自动固相萃取—高效液相色谱双波长法测定现制饮料中7种合成着色剂的方法。方法:样品经纯水提取2次,离心后,合并上清液,调节pH至3~4。利用全自动固相萃取装置活化Poly-sery色素专用固相萃取小柱,并上样、淋洗、洗脱。洗脱液氮吹浓缩至200μL,用50%流动相A+50%流动相B混合液定容至1.0 mL。色谱柱采用C 18柱,流动相为20 mmol/L乙酸铵—甲醇,梯度洗脱,流速1.0 mL/min,进样量10μL,检测波长254,628 nm。结果:各物质在0.5~50.0μg/mL范围内相关系数均大于0.9999,方法检出限为0.023~0.179 mg/kg,定量限为0.078~0.598 mg/kg,加标回收率为92.4%~96.8%,相对标准偏差(RSD)为1.2%~4.2%。结论:该方法试剂使用量小,操作步骤简便,自动化程度高,回收率高,重复性好,适合于批量样品测定。

宠物用丹参咀嚼片制备工艺研究及质量标准建立

旨在确定宠物用丹参咀嚼片的制备工艺并建立其质量标准。本试验通过湿法制粒辅料种类、制粒工艺、压片工艺进行考察,确定其制备工艺,采用薄层色谱法、高效液相色谱法(HPLC)对宠物用丹参咀嚼片中丹参酮ⅡA、丹酚酸B分别开展定性和定量研究,建立其质量标准。结果:将丹参喷雾粉加辅料混合均匀,混合时间5 min,转速100 r/min,以10%淀粉浆为黏合剂制成颗粒,干燥温度40~50℃,0.3%硬脂酸镁作为润滑剂,压片速率10~35 r/min;薄层鉴别显示丹参酮ⅡA斑点清晰,分离度良好且专属性强;高效液相色谱显示丹酚酸B在8.0~256.0μg/mL范围内线性关系良好(R^(2)=0.999 9),系统适用性、专属性、精密度、稳定性、重复性、系统耐用性良好,加样回收率为95.0%~105.0%;对6批样品中丹酚酸B进行含量测定,设定含量限度为11.484 1 mg/g。综上,本试验确定的宠物用丹参咀嚼片制备工艺稳定可行,建立的薄层鉴别方法和高效液相色谱含量测定方法可用于宠物用丹参咀嚼片的质量控制。

清汤药膳羊肉工艺优化及杀菌后品质控制

甘味预制菜清汤药膳羊肉不仅具有独特风味,而且能起到预防疾病和增强免疫力的作用。以甘味预制菜清汤药膳羊肉为研究对象,利用响应面法优化清汤药膳羊肉的工艺参数,同时比较不同杀菌条件对清汤药膳羊肉品质的影响,以优化杀菌工艺。结果表明:以200 g羊肉为基准,在黄芪添加量为0.8%、党参添加量为0.6%、食盐添加量为3.5%、熬制时间为45 min条件下,清汤药膳羊肉的口感、滋味最佳。通过比较以最佳工艺生产的清汤药膳羊肉在4种杀菌条件处理后的理化和感官指标,筛选出最适杀菌条件为108℃/30 min。利用气相色谱仪检出经过最适杀菌条件处理的清汤药膳羊肉中存在14种脂肪酸,其中含多种对人体有益的饱和脂肪酸,如顺亚油酸、花生四烯酸、硬脂酸等,该数据对预制菜工业化生产具有一定指导意义。

低渗储层渗吸剂界面性能与组分分离

对石油磺酸盐WPS-D和十八烷基二甲基甜菜碱复配的渗吸剂进行界面张力测试,得出0.3%浓度8∶2比例下的渗吸剂达到超低界面张力(<1.0×10^(-3)mN/m),0.4%和0.5%浓度比例为9∶1的体系接近超低界面张力。采用制备色谱与质谱相结合,对石油磺酸盐WPS-D进行组分分离,并结合界面张力测试结果,考察不同组分的界面活性。结果表明,石油磺酸盐分离出5种结构组成差异较大的不同组分;以重烷基苯磺酸盐为主,C_(17)~C_(19)烷基苯磺酸盐达到超低界面张力并维持,C_(18)~C_(24)的烷基苯磺酸盐接近超低界面张力后反弹,得出提供界面活性的主要组分是C_(17)~C_(19)磺酸盐。

制备高效液相色谱用于纽莫康定杂质B1和B5的纯化

目的开展低含量成分的富集和纯化研究,从Glarea lozoyensis菌体的发酵产物(即纽莫康定B0粗品)中制备出杂质B1和B5,并对其进行结构鉴定。方法建立了基于制备高效液相色谱(prep-HPLC)技术的两步纯化法,第一步使用亲水(HILIC)模式固定相对目标杂质进行富集,第二步采用反相(RPLC)模式固定相对杂质进行制备。得到的两个杂质采用质谱(MS)、核磁(NMR)进行鉴定,确定化合物结构。结果两个杂质为B1和B5,相对分子质量均为1049 Da,色谱纯度分别为97.83%和98.13%。经核磁鉴定,B1为B0的高酪氨酸类似物,B5为B0的鸟氨酸类似物。结论本研究发展的基于prep-HPLC的两步纯化方法为低含量成分制备提供参考,第一步分离实现目标成分富集,第二步利用正交性的色谱柱提高分离选择性,成功制备出杂质B1与B5,有助于B0杂质谱的建立和进一步的质量控制研究。

基于低温制备色层法的溶解尾气中氪-85纯化装置与方法

^(85)Kr是一种非常有价值的放射性气态核素。目前,我国^(85)Kr气体基本依赖进口满足市场需求。纯化作为^(85)Kr产业提取-纯化一体化生产系统的重要技术,是获得高纯度^(85)Kr的重要手段,因此在进行纯化热实验以及正式的工业生产前需要进行冷实验确定纯化过程的重要参数。搭建了基于低温制备色层法的纯化冷实验台架,研究了色层柱填料和温度对氪气纯化效果的影响,结果表明:椰壳活性炭具有比5A分子筛更好的纯化效果;采用椰壳活性炭填充色层柱,色层柱温度降低至-40℃时,氪气可以和氮气、氧气等杂质完全分离,氪气的纯化效果较好,产品气中氪-84的峰显著高于氮气和氧气,氪气的纯度大幅提高。

利用AutoPrep离子色谱技术实现复杂基体样品中重稀土元素的分离和制备

研究了AutoPrep自动前处理装置和离子色谱联用技术分析复杂基体样品中重稀土元素的分离和半制备的方法。选用两个双层四通阀和一个六通阀替代传统方法中的四个双层四通阀,选用一个GS50四元梯度泵实现对螯合浓缩柱不同条件的淋洗,将吸附或交换在螯合柱和浓缩柱上的碱金属、碱土金属、过渡金属、重金属和重稀土元素选择性地分段洗脱,并将基体消除后的样品在线进样到CS5A离子色谱交换柱中进行最终分离,过柱衍生后进入紫外-可见光检测器进行光度检测,并根据确定的保留时间段进行在线样品收集,完成制备工作。对基体简单且被测组分含量较高的样品,采用直接进样法测定4种重稀土元素(Tb、Y、Tm和Lu)的检测限(S/N=3)为8.3-21.0μg/L,线性相关系数均大于0.9992,分离度均大于4,以峰面积计算的方法精密度(RSD,n=9)小于3%,回收率为89.0%-110.8%;对复杂基体样品或被测组分含量较低的样品,采用基体消除在线浓缩处理,方法的检测限(S/N=3)为0.67-1.43μg/L,线性相关系数均大于0.9994,分离度均大于3,方法的精密度(RSD,n=9)小于3%,回收率为91.2%-106.0%。对简单基体和复杂基体两种样品的分离和制备方法均具备灵敏度高、选择性好、自动化程度高、节省时间等特点,用于岩石等实际样品的检测,结果满意。

黑果分散片的制备、质量及体外活性研究

为了探索黑果分散片的制备工艺、质量及其体外活性,通过单因素与正交试验相结合的方式考察了黑果分散片的制备工艺,采用UV法考察了黑果分散片总三萜的含量及其体外活性,采用超高效液相色谱法测定了黑果分散片中没食子酸、儿茶素、表儿茶素的含量.结果发现黑果分散片的优选处方为微晶纤维素作稀释剂、交联聚乙烯吡咯烷酮作崩解剂、75%乙醇为黏合剂、微粉硅胶作润滑剂.分散片中总三萜的含量测定方法显色前后稳定性符合测定要求、仪器精密度和重复性良好,总三萜的加样回收率为99.48%;体外活性试验结果表明,黑果分散片具有一定的清除DPPH和抑制脂质体的能力;超高效液相色谱法测定没食子酸、儿茶素、表儿茶素的加样回收率分别为99.83%、100.69%和99.61%.优选的制剂工艺制备的分散片,其崩解时限符合要求,总三萜的含量为1.42 mg/片,没食子酸、儿茶素、表儿茶素的含量分别为111.15μg/g、940.19μg/g、598.25μg/g.试验结果为黑果分散片的应用提供了理论依据.

头孢克肟制剂中有关物质结构鉴定与定量分析

目的旨在建立头孢克肟制剂中有关物质结构鉴定与定量分析方法。方法采用HPLC-Q-TOF-MS/MS法对主要杂质结构进行分析。建立HPLC法,采用C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-四丁基氢氧化铵溶液为流动相进行梯度洗脱,检测波长为254 nm,柱温为40℃,对头孢克肟制剂进行有关物质检测。结果初步鉴定了头孢克肟制剂中9个杂质,分别为杂质A、B、D、E、F、G、H、头孢克肟亚砜和头孢克肟叔丁酯。在建立的HPLC色谱条件下,头孢克肟及各杂质均能达到基线分离,各有关物质的质量浓度在0.1934~20.03μg·mL^(-1)内线性关系良好,r>0.9990;回收率为98.4%~101.8%(RSD<2.0%)。结论HPLC-Q-TOF-MS/MS技术能初步鉴定头孢克肟制剂的杂质结构,该方法能检测不同厂家头孢克肟片剂、颗粒剂、胶囊剂及干混悬剂中的有关物质。