Determination of aconitine alkaloids in aconitum roots

A reliable method for extraction of the crude drug, Aconitum roots, and purification of the extract for its eventual separation and quantitation of the aconitine alkaloids by HPLC technique has been developed. The method comprises extraction of aconite powder first with ammoniacal ether and then with methanol (x 3 each), chromatography of the resultant extract over neutral alumina and elution with ethyl acetate-methanol (7:3) prior to HPLC. The method has been found to be reproducible and suitable for routine analysis of aconite and its pharmaceutical preparations.     

复方羊蛇颗粒处方药材提取工艺的优化研究

目的 优化医院制剂复方羊蛇颗粒处方药材的提取工艺。方法 建立处方中有效成分去乙酰车叶草酸甲酯、阿魏酸的高效液相定量方法,并以去乙酰车叶草酸甲酯、阿魏酸的含量以及提取物干浸膏得率为指标,采用星点设计—效应面法对复方羊蛇颗粒处方的浸膏提取工艺进行优化。结果 建立的复方羊蛇颗粒中有效成分的高效液相定量方法符合方法学验证要求。通过星点设计—效应面法优化的最佳提取工艺为：提取溶媒的用量为处方药材量的12倍,醇沉浓度为73%,提取时间为每次60分钟。结论 本研究成功建立了复方羊蛇颗粒有效成分的高效液相色谱定量方法,经优化后的提取工艺操作简单易行、重复性良好。     

白芷香豆素类化合物的成分分析及CO_2超临界萃取工艺的研究

目的:建立以高效液相色谱(HPLC)法分析白芷香豆素类化合物的成分及CO2超临界流体法萃取白芷中香豆素类成分的的方法。方法:采用正交试验设计考察萃取压力、萃取温度、萃取时间及药材粒度对总香豆素收率的影响,用HPLC法检测萃取物中欧前胡素和异欧前胡素的含量。结果:欧前胡素、异欧前胡素的检测浓度线性范围分别为29.4～235.2、10～80μg/ml,平均回收率分别为(100.46±1.42)%、(99.94±1.18)%(n=3),相对标准差分别为1.76%、1.62%;以无水乙醇作为改性剂,白芷在萃取压力35MPa、萃取温度45℃、药材粒度60目、萃取时间3h时总香豆素收率最高。结论:萃取压力、药材粒度、萃取时间对总香豆素萃取收率均有显著影响;所建立的分析方法精密度高、简便快速,适于欧前胡素和异欧前胡素的含量测定。     

乌头中主要生物碱的高效液相色谱测定

用高效液相色谱分离、测定乌头及附子中一些主要生物碱。生药的乙醚提取物在十八烷基键合相柱上,用甲醇—水—氯仿—三乙胺(70:30:2:0.1)作流动相,中乌头碱、乌头碱及次乌头碱与其它杂质能很好分离。以β-甲基萘作内标化合物,用峰高比测定各生物碱含量。曾测定了不同品种、不同产地及不同加工炮制方法的一些样品,其生物碱组成及比例相差较大。     

Column-Switching High-Performance Liquid Chromatographic (HPLC) Determination of Hydrochlorothiazide in Rat,Dog, and Human Plasma

A fully automated HPLC assay for hydrochlorothiazide in plasma has been developed using a column-switching technique. The method involves direct injection of plasma to the extraction column for sample cleanup followed by switching onto the analytical column. Good precision, accuracy, and linearity were obtained over a range of 25 to 2000 ng/ml in rat, dog, and human plasma. The column-switching method has also been validated by comparison with a conventional HPLC method requiring a cumbersome plasma extraction procedure. Since the method is simple, rapid, and reproducible, it is useful for determination of hydrochlorothiazide levels in animal and human plasma.     

Quantification of active principles and pigments in leaf extracts of Isatis tinctoria by HPLC/UV/MS

An HPLC method has been developed and validated for the quantification of the pharmacologically active principles tryptanthrin (1), 1,3-dihydro-3-[(4-hydroxy-3,5-dimethoxyphenyl)methylene]-2 H-indol-2-one (indolinone) (3), indirubin (4), alpha-linolenic acid (2), and indigo (5), an isomer of indirubin, in extracts from the traditional anti-inflammatory plant Isatis tinctoria (woad). The chromatographic separation was performed on a C-18 column with a linear gradient of acetonitrile in water containing 0.1% formic acid. The method combines UV and electrospray MS detection in the positive ion mode for the detection of the alkaloids, with a switch to the negative mode for the analysis of alpha-linolenic acid. The method was applied to the analysis of woad extracts obtained by supercritical fluid (SFE) CO2 extraction, and by pressurized liquid extraction (PLE) with dichloromethane and methanol, respectively. While the highest concentration of alpha-linolenic acid was found in the SFE extract (7.43%), the concentrations of tryptanthrin , indolinone, indirubin and indigo were the highest in the dichloromethane extract (0.30, 0.035, 2.48 and 0.84%, respectively). Compound 3 was not detected in the methanolic extract and only traces of compounds 1, 4 and 5 and low amount of alpha-linolenic acid (0.39%) were present in this extract.     

海娜花抗真菌有效成分的提取工艺优化与抑菌效力研究

目的 研究海娜花抗真菌有效成分的提取工艺优化与抑菌效力。方法 分别采用回流提取法与超声提取法提取海娜花。采用高效液相色谱作定量分析,流动相A为磷酸二氢铵30 mmol/L、B为甲醇（用磷酸调pH=3）,梯度洗脱,温度为30℃,进样量为20μL,色谱柱为Agilent Zorbax SB-C18柱（250 mm×4.6 mm,5μm）。考察溶剂、温度、时间对有效成分提取效果的影响,确定最佳提取工艺。采用美国临床实验室标准化委员会（NCCLS）产孢丝状真菌抗真菌药敏试验方案M38-A,以白色念珠菌为模型菌,对海娜花提取物的抗真菌效力进行初步研究。结果与结论 回流提取法的最佳工艺条件为10倍量70%乙醇溶液,100℃下回流提取1.0 h,共提取3次。超声提取法的最佳工艺条件为10倍量70%乙醇溶液,50℃下超声提取1.5 h,共提取3次。海娜花提取物对白色念珠菌的最低抑菌浓度（MIC）为320～640μg/mL。     

反相高效液相色谱法测定参附注射液及市售黑附片中乌头碱的含量

目的　测定参附注射液及市售黑附片中乌头碱的含量。方法　用RP HPLC ,流动相为乙腈 醋酸铵缓冲液 (pH10 5 ) ,紫外检测波长 2 3 0nm ,流速 0 8ml·min-1。结果　对照品的线性范围 1 3 9～ 2 2 2 4 μg(r =0 9997) ;市售黑附片中的乌头碱比参附注射液中的高 3倍。结论　方法简便、快速、准确、重现性好 ,可以作为样品的检验方法     

3种乌头类中药在大鼠体内外的神经毒性

目的 考察几种常见乌头类中药的神经毒性.方法 采用体内、外给药的方法,大鼠连续90 d经口给予生川乌和生草乌,检测其对大鼠神经行为、血液、血液生化及脑组织的影响;采用大鼠胚胎海马神经元体外培养的方法,研究生川乌、生草乌和生附子对神经细胞的体外毒性.结果 经口给予生川乌和生草乌后,大鼠的神经行为、血液与血液生化指标无变化,脑组织检查未见异常;体外试验结果表明,生川乌、生草乌及生附子对大鼠海马神经元具有一定毒性,呈剂量效应关系,毒性强度为生草乌＞生川乌＞生附子.结论 乌头类中药在大鼠整体动物试验中未见神经毒性作用,但在海马神经元体外培养模型中,表现出神经毒性,这可能与血脑屏障及体内代谢有关.     

仙鹤草中鹤草酚的提取及分离纯化研究

目的：从中药仙鹤草中提取鹤草酚,并对其初步分离纯化进行研究。方法采用超声波法对仙鹤草活性成分鹤草酚进行粗提,并依次用两相溶剂萃取法与酸碱分离法进行纯化。采用薄层层析法和紫外分光光度法对其纯度进行定性分析;用高效液相色谱对其进行含量测定。结果分离纯化的鹤草酚纯度达到65．09％。结论研究出了一条以仙鹤草为原料,经超声波提取,再经乙酸乙酯、碳酸钠、及氢氧化钠酸碱分离,纯化鹤草酚的新工艺路线。     

生桃仁中苦杏仁甙提取工艺研究

目的 　研究生桃仁中苦杏仁甙的提取工艺条件。方法 　通过四因素三水平的正交试验 ,采用 HPLC法测定苦杏仁甙 ,经方差分析优化筛选苦杏仁甙的提取工艺条件并考察其稳定性。结果 　不同提取方法对生桃仁中苦杏仁甙的提取率有极显著影响 (P<0 .0 1) ,采用 90℃以上热水直接继续加热沸提 ,提取物中苦杏仁甙含量是冷水浸泡后再加热沸提的 2倍。水用量、提取次数及提取时间对苦杏仁甙提取率无显著影响 (P>0 .0 5 )。 结论 　生桃仁中苦杏仁甙的优化提取工艺条件为直接加 10倍量沸水提取 1次 ,提取时间为 2 h,工艺稳定可行     

Determination of saponosides from Panax ginseng C.A. Meyer. Comparison of official methods and HPLC

Official methods for determination of ginsenosides of the French and Helvetic Pharmacopoeias have been compared with HPLC method. Sample preparation schemes used are those of monographs with conventional solvent extraction and solid phase extraction with a polar and a non-polar sorbents, respectively kieselguhr and C 18 octadecyl. Liquid-solid sample clean-up with C 18 cartridge is the most effective procedure. Prior HPLC method, an hydrolysis step of malonylginsenosides is necessary. Very selective extraction resulting in highly purified solution authorizes reliable and rapid colorimetric determination from ginseng saponosides.     

穿心莲药材指纹图谱提取方法研究

目的以建立指纹图谱为目标,研究不同提取方法对穿心莲药材化学成分提取效果的影响。方法采用不同提取方式、提取时间、提取溶剂及提取温度对穿心莲药材化学成分进行提取,并对提取物进行高效液相色谱法检测比较。结果以75%甲醇水溶液作为提取溶剂,80℃水浴回流2h的提取条件下获得的指纹图谱信息较为丰富,响应值较高。结论此方法简便、重现性好,可作为构建穿心莲药材HPLC指纹图谱的供试品溶液制备方法。     

翻白草中槲皮素的提取工艺优化及含量测定

目的:优选翻白草中槲皮素的最佳提取工艺,建立以高效液相色谱法测定其含量的方法。方法:采用超声波提取法,以槲皮素的提取含量为评价指标,以提取时间、料液比、乙醇浓度和提取次数为考察因素,通过正交试验筛选出最佳提取工艺;采用高效液相色谱法测定槲皮素的含量。结果:翻白草中槲皮素的最佳提取工艺为每次提取40min、料液比1∶15、80%的乙醇、提取3次;槲皮素检测浓度在0.004～0.020mg·mL-1(r=0.999 6)范围内与其峰面积积分值呈良好的线性关系,平均加样回收率为100.17%,RSD=2.27%(n=6)。结论:本方法简便、迅速,可为翻白草的进一步开发利用和有效性研究提供理论依据。     

正交实验优选苹果皮中根皮苷提取工艺

目的建立苹果皮中根皮苷的提取工艺。方法以红富士苹果为试材,乙醇作为提取溶剂,分别使用浸提法、回流法、超声波法和微波法提取苹果皮中的根皮苷,并利用高效液相色谱测定根皮苷的含量。采用L16（43）正交表确定最佳提取工艺。结果回流法提取效果较好,40%乙醇溶液、料液比1∶30、回流1 min为优化提取方案。结论本工艺条件对苹果皮中根皮苷的提取率高且稳定,为苹果的药物综合利用提供了依据。     

正交试验优选黄连提取工艺

目的:优选黄连最佳提取工艺.方法:采用正交试验法,以黄连提取的收膏率和盐酸小檗碱提取量为考察指标,利用HPLC法测定盐酸小檗碱的含量.结果:最佳提取工艺为黄连粗粉加8倍量60%的乙醇回流提取3次,每次1 h.结论:优选得到的工艺简便易行,稳定性好.     

Automatic determination of diltiazem and desacetyldiltiazem in human plasma using liquid-solid extraction on disposable cartridges coupled to HPLC--Part I: Optimization of the HPLC system and method validation.

A sensitive and automated method for the analysis of diltiazem and desacetyldiltiazem in plasma has been developed using liquid-solid extraction (LSE) on disposable extraction cartridges (DECs) in combination with HPLC. After isolation from plasma, the analytes are separated on a highly deactivated octyl silica column with a mobile phase of methanol-0.05 M phosphate buffer (pH 7.4) (62:38, v/v). The analytes are monitored photometrically at 238 nm. The complete preparation of the plasma sample as well as the injection of the final extract on to the analytical column are performed automatically by means of a sample processor equipped with a robotic arm to which is attached a needle dispensing the different liquids. The internal standard solution is first added to the plasma sample. The DEC is then conditioned successively with methanol and phosphate buffer (pH 7.4). A 1.0-ml volume of sample containing the internal standard solution is applied on an extraction cartridge filled with cyanopropyl silica (50 mg). After the DEC has been washed with the same buffer, the analytes are eluted with 0.16 ml of methanol. A 0.14-ml volume of buffer is then passed through the DEC and 0.25 ml of the final extract is injected onto the HPLC column. The absolute recoveries of the drugs are about 90% and the limit of detection for diltiazem is 0.8 ng ml-1. Relative standard deviations of 2.6% (within-day) and 3.7% (between-day) have been obtained for this compound at a plasma concentration of 50 ng ml-1.     

关于《药典》2000版天麻中天麻素测定提取方法的商榷

目的比较现行版药典载天麻中天麻素测定提取方法和加热回流法及超声法,以选择最佳提取方法。方法按药典方法提取,并与加热回流法及超声法比较。结果用水加热回流提取法得率最高。结论水加热回流提取法最佳。     

参花妇康胶囊中丹参醇提取工艺研究

目的：利用高效液相色谱法优化参花妇康胶囊中丹参醇提取工艺。方法以干膏率及干膏中含丹参酮ⅡA 的量为指标,通过正交试验筛选最佳提取工艺。结果最佳工艺条件为乙醇浓度70%,溶剂量6倍,提取时间3 h,提取次数2次。结论醇提法可用于丹参中有效成分的提取。     

Microdialysis sampling of the isothiazolone, PD-161374, and its thiol and disulfide metabolites

A method based on microdialysis sampling combined with high-performance liquid chromatography (HPLC) has been developed for monitoring the anti-HIV agent PD-161374 (isothiazolone) and its thiol and disulfide metabolites in blood. It was demonstrated that unlike blood withdraw and extraction, microdialysis sampling can preserve the distribution among the isothiazolone and its metabolites in blood. The use of a narrow-bore HPLC system, combined with the relatively high probe extraction efficiency (approximately 50%) from the flexible probe design in this work, allows the direct and quantitative determination of the drug and its major metabolites at submicromolar level.