固相萃取结合薄层色谱法鉴别滋肾育胎丸中的何首乌、续断、巴戟天和白术

目的:结合固相萃取法建立滋肾育胎丸中的何首乌、续断、巴戟天和白术的薄层色谱鉴别方法。方法:采用C18固相萃取法,将供试品甲醇超声提取、蒸干、复溶后上样至活化后的固相小柱中,先后用水和不同浓度甲醇洗脱,洗脱速度为1 mL·min^-1。收集水洗脱液,蒸干复溶作为巴戟天鉴别供试品溶液,以三氯甲烷-甲醇-水-冰醋酸(7∶3∶1∶1)为展开剂;收集60%甲醇水洗脱液,蒸干复溶作为续断鉴别供试品溶液,以正丁醇-醋酸-水(4∶1∶5)的上层溶液为展开剂;收集80%甲醇水洗脱液,蒸干复溶作为何首乌鉴别供试品溶液,以环己烷-二氯甲烷-乙酸乙酯-甲酸(10∶1∶1∶0.1)为展开剂;收集100%甲醇洗脱液,蒸干复溶作为白术鉴别供试品溶液,以正己烷-乙酸乙酯-甲酸(40∶10∶0.5)为展开剂。结果:供试品溶液在与对照药材或对照品相应的位置上,显示相同颜色的荧光斑点,阴性样品无干扰。结论:固相萃取可以有效的将滋肾育胎丸中含量低专属性成分进行富集分析,所建立的方法具有操作简单、重复性好等特点,可用作滋肾育胎丸的薄层色谱鉴别。     

高效液相色谱法测定萝芙木中育亨宾和利血平的含量

目的建立检测萝芙木中育亨宾和利血平含量的高效液相色谱分析方法。方法采用C18色谱柱,流动相A为甲醇-水-三氟乙酸(30∶70∶0.1),流动相B为甲醇-水-三氟乙酸(55∶45∶0.1),阶段洗脱,流速为1 mL/min,检测波长为230 nm。结果育亨宾、利血平在10~200μg/mL浓度范围内呈良好的线性关系,加样回收率、精密度、稳定性、重复性良好。结论此法灵敏准确,可用于萝芙木中育亨宾、利血平的含量测定。     

HPLC法测定育亨宾树皮中育亨宾的含量

目的:建立以高效液相色谱法测定育亨宾树皮中育亨宾含量的方法。方法:色谱柱为C18,流动相为甲醇-磷酸盐缓冲液-三乙胺(30∶70∶0.5,pH5.0),流速为1.0mL.min-1,检测波长为270nm,柱温为室温。结果:育亨宾检测浓度的线性范围为10～1000μg.mL-1(r=0.9998);平均回收率为96.5%,RSD=1.23%。结论:本方法操作简便,结果可靠,可用于育亨宾的含量测定。     

高速逆流色谱分离纯化木芙蓉叶中芸香苷

目的建立高速逆流色谱(HSCCC)分离纯化木芙蓉叶中芸香苷的方法。方法采用水提取法制备芙蓉叶粗提物,应用大孔吸附树脂以60%乙醇为洗脱液对粗提物进行洗脱,达到去除杂质和获得黄酮类物质的目的,应用高速逆流色谱仪,以乙酸乙酯∶正丁醇∶水(1∶0.3∶1.5)上相为固定相,下相为流动相,组成溶剂体系对芙蓉叶提取物进行分离,从而获得高纯度的芸香苷。结果经对提取的芸香苷粗提物及HSCCC法纯化后的峰样品进行高效液相色谱(HPLC)分析,表明芸香苷粗提物的纯度较低,纯化后的峰样品纯度98.6%,达到了化学对照品的要求。结论该方法重复性好,分离效果好,可为HSCCC在其他植物黄酮类提取中的应用提供有价值的参考。     

高效液相色谱法测定水飞蓟提取物中水飞蓟宾的含量

目的建立一种用反相高效液相色谱测定水飞蓟提取物中水飞蓟宾含量的方法。方法以Shim-packVP-ODS(250mm×4.6mm,5μm)为分析柱;以甲醇-水-乙腈-磷酸(45∶50∶5∶0.5,体积比)为流动相。结果测得平均回收率为102.3%,RSD为0.75%。结论方法准确可靠、简便,适用于水飞蓟提取物中水飞蓟宾的含量测定。     

重组人红细胞生成素大规模制备中反相色谱的条件优化

目的优化重组人红细胞生成素(rhEPO)大规模制备纯化中主要影响回收率的反相色谱的条件,提高回收率。方法采用反相纯化作为三步法纯化的第一步,利用R1型反相填料采用不同分步洗脱条件进行洗脱。结果优化后经第一步反相色谱所得rhEPO蛋白纯度达70%,再通过阴离子交换和凝胶过滤色谱纯化,rhEPO终产物的纯度可达100%,体内比活达1.91×105IU/mg,rhEPO蛋白的回收率提高了1.2倍。结论优化的反相纯化条件(30%乙醇-1.5 mol/L尿素,60%乙醇-3.0 mol/L尿素)能更有效的从培养上清中大量纯化出rhEPO,回收率高。获得的产品纯度高,体内外活性好。适合大规模纯化。     

从催吐萝芙木中提取利血平和育亨宾的工艺研究

研究从催吐萝芙木中提纯利血平和育亨宾的工艺.催吐萝芙木根粉用酸性乙醇水溶液浸提,正交实验法确定浸提条件为:浸提2次、每次3 h、浸提剂含50%乙醇、0.05 mol/L硫酸.浸提液经HZ-818大孔吸附树脂吸附,用40%乙醇溶液(含1%HCI)解吸育亨宾,用80%乙醇溶液(含1%HCI)解吸利血平,育亨宾和利血平得到完全分离.两种解吸液分别经过沉淀、结晶,得到利血平和育亨宾.所得利血平纯度为99.8%,总收率为27.1%;育亨宾纯度为4.9%,总收率为9.8%.本工艺能从催吐萝芙木中提取利血平和育亨宾,有利于充分利用中药材,降低成本.     

中压制备液相色谱法分离纯化新藤黄酸

目的优化制备型液相色谱PR-HPLC分离纯化新藤黄酸的方法。方法以新藤黄酸得率为考察指标,藤黄醇提取粗品用PR-HPLC分离纯化,正交设计法优化分离条件,HPLC进行含量测定。结果最佳制备色谱分离条件为:上样量50 g,流速180 ml.min-1,流动相甲醇-水(80∶20),室温进行;所得新藤黄酸为明黄色针状结晶,纯度不小于95%。结论工艺优化合理,重现性好。     

萝芙木中育亨宾大鼠体内药物动力学

目的建立育亨宾在大鼠体内的高效液相色谱测定方法,测定萝芙木中育亨宾在静脉注射后大鼠体内的药物动力学行为。方法采用大连中汇达Hypersil-C18(250 mm×4.6 mm,5μm)色谱柱,以甲醇-体积分数为0.05%的三乙胺水溶液(体积比为75:25)为流动相,流速为1.0 mL.min-1,检测波长为280 nm。大鼠静脉注射育亨宾1 mg.kg-1后,于不同时间点眼眶采血,HPLC法测定其血药浓度,经DAS2.1.1版药动学软件处理得到药动学参数。结果大鼠血浆中育亨宾的线性范围为0.10～3.00 mg.L-1(r=0.995 2),定量下限为0.10 mg.L-1。tmax为0.083 h,ρmax为(2.095±0.302)mg.L-1,AUC0-t为(2.423±0.417)g.h.L-1,AUC0-∞为(2.634±0.412)g.h.L-1。Cl为(0.389±0.072)L.h-.1kg-1。结论建立的大鼠血浆中育亨宾含量的HPLC测定方法适合萝芙木中育亨宾大鼠体内的药物动力学研究;育亨宾静脉注射后消除较快。     

高纯度平阳霉素铜螯合物纯化工艺研究

本试验采用单分散聚合物微球填料柱色谱法纯化平阳霉素铜螯合物粗品,使其HPLC纯度高于98%。首先采用XR 3SP微球反相色谱纯化,使纯度和单步收率分别提高至50%和85%以上。然后经过POlyRP 30-300反相色谱纯化,纯度和单步收率可分别提高至98%和88%以上,总收率达75%以上。该工艺简单可靠,适于工业化生产。     

Synthesis and identification of pilocarpic acid diesters, prodrugs of pilocarpine.

A series of new pilocarpic acid diesters were synthesized to obtain prodrugs for pilocarpine with varying physico-chemical properties. Thermospray liquid chromatography-mass spectrometry (TSP-LC-MS), liquid chromatography with UV-detection (LC-UV) and NMR-spectroscopy were used for the identification of the synthetic products and for evaluation of their purity including typical impurities (pilocarpic acid monoester, pilocarpine). TSP-LC-MS-analysis was performed in the reversed-phase mode using acetonitrile (60%)-0.2 M ammonium acetate (40%) as mobile phase. In LC-UV-analysis chromatographic separation was carried out on a reversed-phase column and the mobile phase consisted of methanol (71%) and 0.02 M potassium dihydrogen phosphate, pH 4.5 (29%). Electron ionization-mass spectrometry (EI-MS) was used for elucidation of structures. Elemental compositions of the substances were verified with high resolution-mass spectrometry (HR-MS). The complete establishment of structures presented was based on 1H-, and COSY-NMR-spectroscopy joined to TSP-LC-MS-analysis.     

吡柔比星的分离与纯化

目的确定用硅胶柱层析纯化吡柔比星粗品的工艺条件。方法采用薄层层析-硅胶柱层析法考察不同展开剂的层析效果,最优洗脱剂为二氯甲烷-甲醇;以吡柔比星的回收率和平均含量为评价指标,考察了流速、进样量与层析柱再生条件对层析粗分和精分的影响。结果硅胶柱层析粗分吡柔比星的最佳工艺条件:以梯度洗脱方式洗脱,洗脱剂流速5.0mL.min-1,上样量16.6mg.mL-1柱体积,0.5g.L-1氢氧化钠溶液离线再生硅胶。吡柔比星质量分数经粗分后由原来的35.7%提高到90%左右,回收率为96.2%。硅胶柱层析精分吡柔比星的最佳工艺条件为洗脱剂二氯甲烷-甲醇比例为90∶10,流速3.5mL.min-1,上样量16.6mg.mL-1柱体积,甲醇再生3倍柱体积。吡柔比星质量分数经精分后由粗分的90%提高到99.6%左右,回收率为97.5%。结论硅胶柱层析纯化吡柔比星所得产品符合质量要求。     

Liquid chromatography in pharmaceutical analysis IV: determination of antispasmodic mixtures.

Parameters associated with the separation of antianxiety-antispasmodic agents were investigated using high-pressure liquid chromatography. Eight widely prescribed drugs were studied. The compounds were chromatographed on reversed-phase octadecyltrichlorosilane (C18) or diphenyldichlorosilane (phenyl) columns, using mixtures of absolute methanol and distilled water buffered with ammonium dihydrogen phosphate, ammonium acid phosphate, or ammonium carbonate. A mixture of phenobarbital-propantheline bromide was selected to demonstrate the utility of the separation and quantification method. The mixture was chromatographed on a phenyl column, using absolute methanol-aqueous 1 percent ammonium dihydrogen phosphate (60:40) (pH 5.85) at a flow rate of 1.4 ml/min. Each determination can be achieved in approximately 15 min with an accuracy of 1-2 percent.     

Characterization of drug purity by liquid chromatography

Applications of liquid chromatography (LC) in the characterization of drug purity are reviewed. Examples are given of the development of methods for the separation of impurities in acids, amines and neutral drug substances. A variety of methods and techniques are discussed including normal-phase and reversed-phase LC, gradient elution, and multidimensional separation systems.     

Statistical optimization of a reversed-phase liquid chromatographic method for the analysis of amiloride and hydrochlorothiazide in tablets

A method has been developed for the separation of hydrochlorothiazide and amiloride by high-performance liquid chromatographic (HPLC) method on a C18 column with detection at 280 nm. The optimal conditions of separation were determined with the aid of 'window diagram' technique of Laub and Purnell. The effect of simultaneously varying the pH, proportion aqueous acetic acid and methanol in the mobile phase were studied to optimize the separation. A response surface diagram was used to optimize the experimental conditions for the separation. The mobile phase composition that provides an acceptable resolution hydrochlorothiazide and amiloride in a short elution time is water:methanol (60:40) and pH 3.2 (pH adjusted to 3.2 with CH3COOH). A method is applied for the quantitative analysis of Moduretic tablets (Merck Sharp & Dokme International). The powdered tablets are extracted with methanol, containing caffeine as the internal standard, and assayed by comparison of peak areas after liquid chromatography.     

反相高效液相梯度洗脱法测定人血浆中丙泊酚浓度

目的：建立高效液相色谱法测定人血浆中丙泊酚血药浓度的方法,为保证临床用药的有效性和安全性提供依据。方法以卡马西平为内标,色谱柱为Hanbon Science ＆．Technology C18（4．6 mm ×250 mm,5μm）,流动相以乙腈／甲醇混合液（乙腈—甲醇按80∶20（V／V）的比例配制,作为有机相）—水（水相加1％的三氟乙酸调pH值到4）进行二元梯度洗脱,0～7 min （50∶50,V／V）,7～9 min（50∶50～70∶30,V／V）,9～22 min（70∶30,V／V）;流流速为1．0 mL· min－1,检测波长为274 nm,进样量为20μL,柱温为40℃。结果丙泊酚的血药浓度在0．098～25 mg· L－1内线性良好（r＝0．9998）,定量限为0．098 mg· L－1;高（12．50 mg· L－1）、中（1．56 mg· L－1）、低（0．19 mg· L－1）3个浓度的平均方法回收率均在85％～115％之间,相对标准偏差（ RSD）均＜10％;日内、日间RSD均＜10％。结论该方法灵敏、准确、简单,可用于丙泊酚的血药浓度监测和临床药动学研究。     

不同比例川芎当归共煎液指纹图谱考察

目的:考察不同比例川芎、当归共煎液指纹峰的变化情况。方法:采用高效液相色谱法,以ZorbaxSB-C18(250mm×4.6mm,5μm)为色谱柱,以甲醇-0.5%醋酸水溶液为流动相进行梯度洗脱,检测波长为313nm。结果:不同比例川芎、当归共煎液中未发现新峰生成;川芎、当归指纹峰具有加合性,但并非简单加合,某些成分的煎出量受配比的影响,川芎∶当归=7∶3时阿魏酸煎出量最大。结论:本研究可为抗心血管药物的开发提供思路。     

高速逆流色谱法分离纯化长春花中长春新碱

目的采用高速逆流色谱(HSCCC)分离纯化长春花中长春新碱,建立相关工艺条件,为工业生产提供参考数据。方法采用高速逆流色谱仪,氯仿-甲醇-0.3mol.L-1 HCl(4∶3∶2)溶剂体系,上相为固定相,下相为流动相,流动相流速为2.0mL.min-1,紫外检测波长为280nm,主机转速为800r.min-1,恒温循环温度为28℃。结果 HPLC检测HSCCC纯化后长春新碱质量分数为79.7%。结论高速逆流色谱纯化长春花中长春新碱容量大、分离纯度和效率高,避免了有机溶剂的大量使用,适用于工业生产高纯度产品。     

High performance liquid chromatographic enantiomer separation of beta-blocking agents with a new isothiocyanate type chiral derivatizing agent

The applicability of (1R,2R)-1,3-diacetoxy-1-(4-nitrophenyl)-2-propyl isothiocyanate [(R,R)-DANI] as a recently developed chiral derivatizing agent for the enantioseparation of a series of beta-blockers is described. The thiourea diastereomers formed were analysed by reversed-phase high-performance liquid chromatography, mixtures of water and methanol or acetonitrile being used for elution. Conditions of derivatizations (temperature, reagent excess and reaction time) were optimized. The effects of organic modifiers on the retention and separation were investigated; the diastereomers could readily be baseline-separated with methanol-containing mobile phases. The reagent allowed the detection of all four stereoisomers of labetalol, which has two chiral centres.