非水液相微萃取-高效液相色谱法测定大黄中5种游离蒽醌类化合物

目的利用非水液相微萃取-高效液相色谱法(HPLC)同时测定大黄中游离蒽醌类化合物的含量。方法利用自制的液相微萃取装置,以聚偏氟乙烯中空纤维为溶剂载体,正己醇为萃取溶剂,供相为甲醇,接受相为1mmol/LNaOH,搅拌速度为1500r/min,萃取时间为50min。萃取结束,接受相在434nm处进行HPLC分析。结果在优化的液相微萃取条件下,芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚的线性范围、回收率、精密度和检测限分别为:0.015～13.2、0.075～11.2、0.085～12.8、0.096～14.4、0.117～17.6μg/ml;103.8%～106.1%;2.5%～5.2%和2.9～20.0ng/ml。结论本法选择性高,有机溶剂消耗少,能快速、准确地测定大黄药材中游离蒽醌类化合物。     

固相萃取-高效液相色谱法测定银线莲中的核苷酸

目的 建立一种固相萃取-高效液相色谱联合检测银线莲中核苷酸的分析方法.方法 采用固相萃取柱对银线莲中的核苷酸成分进行净化、富集,用高效液相色谱法测定含量.色谱柱为:Agilent TC-C18(4.6 mm×250 mm,5μm);流动相:A-0.008％甲酸/10 mmol·L-1甲酸铵,B-甲醇,梯度洗脱;流速:0...     

固相萃取高效液相色谱法测定氯氮平血药浓度

目的建立高效液相色谱测定人血清中氯氮平(抗精神病药)浓度的方法。方法血清样品上SPE柱,甲醇为洗脱剂,65℃氮气吹干后复溶进样测定。用Hypersil C18色谱柱,流动相为甲醇-水=75∶25(加入0.3%三乙胺,用冰醋酸调节pH=8.0),流速为0.8 mL.min-1,检测波长243 nm。结果氯氮平血清样品线性范围为0.05～10.00 mg.L-1,低、中、高3种不同浓度(0.22,1.08,3.22 mg.L-1)方法回收率分别为102.0%,99.7%,99.4%,日内、日间RSD均小于5%。结论本方法准确性好,灵敏度高,操作简便,可以满足氯氮平血药浓度监测和药物中毒分析的要求。     

固相萃取-高效液相色谱法测定甲氨蝶呤的血药浓度

目的：建立固相萃取高效液相色谱法测定人血浆中甲氨蝶呤的药物浓度。方法：采用Supelclean萃取小柱提取血浆中甲氨蝶呤，以高效液相法测定药物浓度。结果：甲氨蝶呤的保留时间约为11．5min，与血中内源性杂质分离良好。方法的绝对回收率为92．64％，线性范围为0．104～104mg·L-1，当S／N≥3时，甲氨蝶呤最低检出限度可这50μg·L-1结论：本法准确、灵敏、回收率高，可满足血药浓度监测的需求。     

固相萃取-高效液相色谱法测定血浆舒必利浓度

目的 建立舒必利血药浓度的测定方法。方法 采用固相萃取 高效液相色谱法。样品先经Cleanert ODS SPE固相萃取小柱处理。色谱柱：Hypersil CN（4.6 mm×150 mm，5 μm）；柱温：40 ℃；流动相为5 mmol·L-1磷酸二氢钾溶液 乙腈(55：45)；流速：1.2 mL·min^-1；荧光检测波长λex为244 nm，λem为345 nm。结果 线性范围6.76～1 689.60 ng·mL^-1，线性关系良好，最小可测定浓度为6.76 ng·mL-1。绝对回收率平均为80.2%～85.4%，相对回收率98.3%～104.3％，日内及日间RSD均＜5%。结论 采用固相萃取-高效液相色谱法能快速可靠地测定舒必利血药浓度。     

固相萃取-高效液相色谱法测定3种抗癫痫药物血药浓度

目的:建立快速固相萃取-高效液相色谱法测定3种抗癫痫药血药浓度.方法:采用Accubond SPE C18小柱处理血样, Nova-pak C18柱(4 μm,3.9 mm×150 mm)为分析柱;0.01 mol·L-1磷酸盐缓冲液(pH 3.5)-甲醇(50∶50)为流动相;检测波长210 nm;流速1 mL·min-1,柱温25℃.结果:3组分抽提完全,杂质去除较为完全.在该色谱条件下,3种药物分离良好,苯巴比妥、苯妥英钠、卡马西平线性范围为5～40,5～40,2.5～20 mg·L-1,平均回收率分别为100.0%,99.9%,100.2%.日内日间误差RSD均小于5.01%.应用于55例患者监测,并与液液萃取法比较,差异无显著性(P＞0.05).结论:本法快速简便,结果准确,更适合临床常规监测分析.     

在线固相萃取-高效液相色谱法测定血清中的茶碱

目的:建立一个准确、简单、快速的分析人血清中茶碱含量的方法,并应用于临床治疗药物监测。方法:利用双三元液相色谱系统(Ulimate3000系列),经C₁₈MG S-3(20 mm×4.0mm,A3JW)固相萃取柱进行在线固相萃取,使用Acclaim C₁₈(150 mm×4.6 mm,5μm)进行分离,梯度洗脱,检测波长273 nm,柱温30℃。结果:利用在线固相萃取液相色谱法测定血清中茶碱的方法检出限为0.10 mg·L^-1,定量限为0.35 mg·L^-1;线性范围为0.35~20 mg·L^-1,R^2=0.999 8;当加标浓度范围为0.35~10 mg·L^-1时,回收率范围为97.28%~103.1%,RSD为0.52%~1.72%。结论:本方法简便、快速,可用于血清中茶碱的临床血药浓度监测及药动学研究。     

固相萃取-高效液相色谱法测定葛根中葛根素含量

目的 研究固相萃取富集除杂,高效液相色谱法测定葛根样品中的葛根素.方法 葛根样品提取液用Sep- Park-C18固相萃取小柱富集除杂,以Agilent Eclipse C18柱(150 mm×4.6mm,5μm)色谱柱为固定相,甲醇-水(25:75)为流动相,流速为0.5mL/min等梯度洗脱,用紫外检测器检测,根据该色谱图的峰面积定量.结果 平均加样回收率为96.08％,RSD为1.51％(n=6).结论 该方法用于测定葛根中的葛根素,结果令人满意.     

固相萃取-高效液相色谱法测定人血浆中硝苯地平浓度

目的　本文采用固相萃取-高效液相色谱法测定人体血浆中硝苯地平浓度。方法　选择固相萃取柱(Bond Elut,C2 )提取样品,在DiamonsilODS(4.6mm×2 5 0mm ,5 μm)分离柱上,以尼莫地平为内标,乙腈-水(5 2∶4 8,V/V)为流动相,紫外吸收波长2 38nm处检测样品。结果　硝苯地平血浆样品在2～6 0 μg·L-1范围内线性关系良好(r=0 .9998,n =6 ) ,方法回收率接近10 0 % ,日内、日间RSD均<5 %。结论　与以往的分析方法相比,本方法提高了检测硝苯地平血浆浓度的灵敏度     

固相萃取高效液相色谱法测定盐酸氯丙嗪血药浓度

目的:建立测定人血清中盐酸氯丙嗪浓度的高效液相色谱法。方法:以C8柱(Hypersil C8,4.6 mm×200 mm,10μm)为色谱柱,流动相为乙腈-水-三乙胺-冰醋酸(700∶300∶5∶2),流速为0.8 mL.min-1,检测波长254 nm。血清样品上SPE柱,以流动相为洗脱剂,收集洗脱液直接进样测定。结果:盐酸氯丙嗪血清样品线性范围为0.2～13μg.mL-1,低、中、高3种不同质量浓度(0.240 0,0.960 0,3.870 0μg.mL-1)方法学回收率分别为101.82%,94.61%,96.15%,日内、日间RSD均小于7.5%。结论:本方法准确性好、灵敏度高、操作简便,可满足临床盐酸氯丙嗪血药浓度监测需要。     

Distribution and pharmacokinetics of five Rhubarb anthraquinones in rabbits and rats

The main active components of Rhubarb are anthraquinones (AQs), most of which are glycosides and others are free. The concentrations of AQs derivatives (rhein, aloe-emodin, emodin, chrysophanol and physcion) in plasma and homogenate were assayed with a high performance liquid chromatography (HPLC) method. The pharmacokinetic parameters and distribution of Rhubarb AQs in rabbits or rats were studied after administrationof different formulas. Elimination of AQs was fit to a two-compartment model in rats and rabbits. There were no significant difference in the main pharmacokinetic parameters between rhein and AQs in rats. AQs were distributed progressively in the kidney, liver, blood, and heart. The AQs were mainly composed of rhein in vivo and was excreted by the kidney. For formulas that contained Rhubarb, rhein could be used as a probe for in vivo pharmacokinetic studies.     

液相微萃取-后萃取光化学荧光高效液相色谱法测定生物样品中甲氨蝶呤的含量

目的:采用液相微萃取-后萃取光化学荧光高效液相色谱法测定生物样品中甲氨蝶呤的含量。方法:应用自制的液相微萃取装置,在0.05mol.L-1盐酸酸性介质中,甲氨蝶呤以分子状态首先被聚醚砜中空纤维孔壁中的正丁醇萃取,继而被25μL0.05mol.L-1氨水溶液后萃取。后萃取液用等体积的0.2mol.L-1盐酸酸化后,经紫外光照射45min,发生光化学反应,取样20μL进行高效液相色谱分析,在λex=275nm,λem=370nm荧光检测,建立了液相微萃取-后萃取-光化学荧光高效液相色谱法测定生物样品中甲氨蝶呤含量的方法。结果:该方法在血浆和尿液中的浓缩倍数可达20~24倍,线性范围分别为0.05~50mg.L-1和0.01~50mg.L-1,检出限分别为10μg.L-1和5μg.L-1,RSD<11%。通过液相微萃取-后萃取,能有效地去除生物样品中干扰甲氨蝶呤测定的内源性杂质,提高了选择性。结论:该方法将液相微萃取和光化学荧光-高效液相色谱法相结合,为生物样品中甲氨蝶呤的检测提供了一种有效的分离分析方法。     

液相微萃取光化学荧光高效液相色谱法测定生物样品中枸橼酸氯米芬顺反异构体的含量

目的:建立液相微萃取光化学荧光高效液相色谱法测定生物样品中枸橼酸氯米芬顺反异构体的含量。方法:应用自制的液相微萃取装置,对生物样品中的枸橼酸氯米芬顺反异构体萃取后,分离采用 Sinochrom ODS—BP C_(18)色谱柱(200 mm×4.6mm,5μm);流动相为甲醇-水(70∶30,每1L 水含0.25 mL 磷酸、50μL三乙胺);流速为0.8 mL·min~(-1);荧光检测器的激发波长和发射波长分别为255 nm 和378 nm;柱温20℃。结果:利用该方法可以将血浆、尿液和肝脏组织匀浆中的枸橼酸氯米芬顺反异构体同时提取分离,浓缩倍数可达5～7倍;枸橼酸氯米芬在血浆、尿液和肝脏中的线性范围分别为0.05～20μg·mL~(-1),0.02～20μg·mL~(-1),0.04～40μg·g~(-1);检出限分别为20 ng·mL~(-1),10μg·mL~(-1),20 ng·g~(-1);精密度试验的 RSD 小于11.7%。结论:本文将液相微萃取技术应用于生物样品中枸橼酸氯米芬顺反异构体的提取分离,利用高效液相色谱法荧光检测,获得了较好的效果。     

比色法测定逐瘀扶正胶囊中蒽醌类成分的含量

目的：建立测定逐瘀扶正胶囊中蒽 醌类成分含量的方法。方法：利用蒽醌类成分在碱性条件下显红色的特性，将经典的混合碱比色法加以改进后用于测定其含量。结果：改进后的比色法操作简便快捷。结果准确（平均加样回收率为99.86%)，重复性好（RSD=1.58%,n=5)。可有效控制逐瘀扶正胶囊的质量。结论：所建立的混合碱比色法可作为逐瘀扶正胶囊中蒽醌类成分的定量方法，对于其它含有蒽醌类成分中药制剂的定量分析亦具有一定的方法学意义。     

反相高效液相色谱法测定脑脉通有效部位中蒽醌类成分的含量

目的:建立脑脉通有效部位中的蒽醌类成分的反相高效液相色谱法含量测定方法。方法:采用大连依利特ODS色谱柱(250 mm×4．6 mm,5μm),流动相为甲醇-0．1％磷酸溶液(80:20,V/V),流速:1．0 mL·min~(-1);检测波长:254 nm。结果:芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚总含量为4％以上,平均回收率为100．54％,RSD:1．58％。结论:所建立的方法简便、准确,可作为有效部位的质量控制方法。     

紫外分光光度法测定巴戟天配方颗粒中总蒽醌含量

目的测定巴戟天配方颗粒中总蒽醌含量。方法用紫外分光光度法测定其含量,测定波长为425 nm。结果检测浓度在0.208 8～3.132 0 mg/ml范围内与吸收度线性关系良好(r=0.994)。结论研究的方法可有效地控制巴戟天配方颗粒的质量。     

Analysis of rhubarb anthraquinones and bianthrones by microemulsion electrokinetic chromatography

In this work a method of microemulsion electrokinetic chromatography (MEEKC) has been developed for the analysis of nine anthraquinones and bianthrones in rhubarb. This study employed di-n-butyl tartrate as oil substance to make up the microemulsion. The composition of the microemulsion was 0.5% (w/w) di-n-butyl tartrate, 0.6% (w/w) SDS, 1.2% (w/w) 1-butanol and 97.7% (w/w) 10 mM sodium borate buffer, pH of the buffer being 9.2. Acetonitrile was added to the emulsion to improve the separation. The volume ratio between the emulsion solution and acetonitrile of an optimized separation was 70:30. With the optimized conditions all of the nine analytes were baseline-separated in peaks of good shapes within 20 min. After validation the method was used to analyze the components in a rhubarb sample. A solid-phase extraction procedure was employed. Five anthraquinones and two bianthrones had been detected in the sample and their amounts were determined. The method should be able to be used for the quantitative analysis of the main active components of rhubarb crude drugs.     

Determination of five components in Ixeris sonchifolia by high performance liquid chromatography

A high performance liquid chromatography (HPLC) method was developed for the simultaneous quantification of five major active ingredients (markers) in Ixeris sonchifolia (Bge.) Hance, namely chlorogenic acid, caffeic acid, luteolin-7-O-beta-D-glucuronide, luteolin-7-O-beta-D-glucoside and luteolin. Samples were extracted with 70% methanol. The chromatographic separation was performed on a Hypersil ODS(2) column (250 mm x 4.6 mm i.d.; 5 microm) with a gradient of acetonitrile and 0.5% (v/v) aqueous acetic acid, at a flow rate of 1.0 ml/min, detected at 335 nm. Five regression equations showed good linear relationships (r(2)>0.999) between the peak area of each marker and concentration. The assay was reproducible with overall intra- and inter-day variation of less than 3.2%. The recoveries, measured at three concentration levels, varied from 94.1% to 100.7%. This assay was successfully applied to the determination of the 5 bioactive compounds in 18 samples. The results indicated that the developed assay method was rapid, accurate, reliable and could be readily utilized as a quality control method for I. sonchifolia (Bge.) Hance.     

Simultaneous determination of five anthraquinones in medicinal plants and pharmaceutical preparations by HPLC with fluorescence detection

A reversed-phase high-performance liquid chromatography (RP-HPLC) method with fluorescence detection for simultaneous determination of five anthraquinones in Rhubarb collected from nine different locations in China, Polygonum cuspidatum, Polygoni multiflori and three pharmaceutical preparations is proposed and validated. Chromatography was carried out at 25 °C on a Hypersil C18 column with the isocratic mobile phase of methanol–0.1% aqueous formic acid (85:15, v/v) at a flow rate of 1.0 ml/min. The excitation and emission wavelengths were set at 440 and 540 nm, respectively. A comprehensive validation of the method included tests of sensitivity, linearity, precision and accuracy. The linear regressions were acquired with r > 0.999. Satisfactory intra- and inter-day precisions were achieved with R.S.D.s less than 3.95% and the average recovery factors obtained were in the range of 93.2–103.8%.