分散液相微萃取-HPLC法用于大鼠血浆中对香豆酸测定

目的建立了分散液相微萃取与高效液相色谱联用技术测定大鼠血浆中对香豆酸浓度。方法微萃取条件为:20μL 1-己基-3-甲基咪唑六氟磷酸盐离子液体(1-hextyl-3-methylimidazoliumhexafluorophosphate,[C6MIM][PF6])作萃取剂,80μL乙腈作分散剂,0.5 mol.L-1硫酸作酸化剂,萃取时间为1 min。结果在优化的萃取条件下,血浆中对香豆酸质量浓度在0.012～2.400 mg.L-1内,线性关系良好(相关系数r=0.997 5),日内和日间精密度(RSD%)<9%(n=6),相对回收率为96.7%～101.6%,提取回收率为59.0%～70.4%。结论该方法适于血浆中对香豆酸浓度的测定。     

HPLC-MS/MS方法检测人血浆中氨溴索的浓度

目的建立液相色谱-电喷雾串联质谱法测定人血浆中氨溴索的浓度。方法血浆样品经乙醚萃取后,以乙腈-0.1%甲酸（50∶50,v/v）为流动相,采用Inertsil C18柱（150 mm×2.1 mm,5μm）分离,流速为0.2 mL.min-1,通过电喷雾离子化串联质谱,以多反应监测（MRM）方式进行检测。结果氨溴索的线性范围为0.81～413.0 ng.mL-1,最低定量浓度为0.81 ng.mL-1,平均相对回收率在80%～120%,日内和日间变异均〈15%。结论本法简单、快速、灵敏、重现性好,是一种有效的检测人血浆中氨溴索浓度的方法。     

反相高效液相色谱法测定人血浆中氯氮平浓度

目的 :以反相高效液相色谱法测定人血浆中氯氮平的浓度。方法 :血浆样品经碱化后用乙醚萃取 ,醚层用100μl0 1mol/mlKH2PO4 反萃取 ,40μl进样。色谱柱为DikmaDiamonsilC18 柱 ,保护柱为WatersNova -PakC18 柱 ,流动相为0 5%三乙胺溶液 -乙腈 (74∶26,V/V) ,流速为1ml/min ,检测波长为215nm。结果 :测定方法在0 1～20μg/ml范围内线性关系良好 ,萃取回收率在80 3%～84 8%之间 ;日内、日间精密度在3 5%～8 3 %之间 ,最低检测浓度为50ng/ml。结论 :本测定方法快速、灵敏、准确 ,适用于临床治疗药物浓度监测。     

毒鼠强人血浆蛋白结合率的研究

目的 建立人标准血浆中毒鼠强浓度的GC/NPD测定法,测定毒鼠强人血浆蛋白结合率.方法 用平衡透析法测定血浆蛋白结合率,用GC/NPD测定法测定血浆侧毒鼠强浓度.结果 正常人血浆中毒鼠强浓度在100ng/ml、300ng/ml、500ng/ml时的蛋白结合率在12.62%～15.41%之间.结论 毒鼠强与人血浆蛋白率较低,属低度结合,对临床现象的解释有重要意义.     

HPLC法测定人血浆中氯氮平浓度

目的建立测定人血浆中氯氮平浓度的HPLC法.方法血浆经碱化后,用OASIS固相萃取小柱提取血浆中氯氮平,采用反相高效液相色谱法二极管阵列检测器检测.色谱柱为Dikma Diamonsil C18柱(200 mm×4.6 mm,5μm),保护柱为WatersNova-Pack C18柱,流动相为0.5%三乙胺溶液(磷酸调pH=3.0)-乙腈(7228,V/V),流速为1 mL·min-1,检测波长(λ)215 nm.结果本方法在0.10～2.00mg·L-1范围内线性良好(r=0.998 8,n=6).高、中、低浓度加样回收率均在92.1%～105.8%之间,日内、日间RSD在4.1%～8.6%之间,最低检测浓度为5μg·L-1.结论本测定方法适用于临床氯氮平血药浓度监测.     

固相萃取反相高效液相色谱法测定人血浆中多奈哌齐浓度

目的:建立固相萃取反相高效液相色谱法测定人血浆中多奈哌齐浓度的方法。方法:血浆经碱化后,用OASIS固相萃取小柱提取血浆中多奈哌齐,紫外检测器检测,检测波长316nm。色谱柱:Sh im-Pack C18柱(150mm×4.6mm,5μm),保护柱:Sh im-Pack C18柱,流动相:0.5%三乙胺溶液(磷酸调pH3.0)-乙腈(65∶35),流速为1.0mL.m in-1。结果:标准曲线线性范围为5~100mg.L-1(r=0.999 3),萃取回收率在81.9%~90.7%之间。日内、日间RSD在5.4%~10.2%之间,最低检测浓度为2mg.L-1。结论:本测定方法快速、灵敏、准确,适用于临床治疗药物浓度检测的要求。     

高效液相色谱-电喷雾质谱法测定人血浆克拉霉素浓度

目的 建立测定人血浆中克拉霉素浓度的高效液相色谱-电喷雾质谱(HPLC-MS/ESI)法。方法 血浆样本经处理后,直接在C₁₈色谱柱上用含甲酸的流动相洗脱,洗脱物通过ESI接口,以正离子模式电离,质谱仪以选择离子检测方式检测定量。结果 血浆克拉霉素浓度在18.0～4 000.0 ng·mL^-1之间线性关系良好,日内变异和日间变异在5.0%之内,回收率94.9%～104.3%。最低检测限18.0 ng·mL^-1。结论 该方法操作简便、快速、准确,适用于克拉霉素的人体药动学研究.     

液相微萃取-非水后萃取-高效液相色谱法测定大鼠体内厚朴酚与和厚朴酚的浓度

目的:建立液相微萃取-非水后萃取(LPME/NBE)-高效液相色谱法同时测定大鼠体内厚朴酚及和厚朴酚浓度的方法.方法:利用自制的液相微萃取装置,以正丁醇为萃取溶剂,以600 r·min-1转速萃取10 min,对大鼠血浆、肝脏和肾脏生物样品中的厚朴酚与和厚朴酚进行分离、萃取、纯化.色谱条件:ODS为色谱柱,甲醇-水(82∶18)为流动相,294 nm为检测波长.结果:厚朴酚与和厚朴酚线性范围分别为0.15～30.0 mg·L-1和0.10～30.0 mg·L-1,r＞0.900;RSD＜7.5%;平均回收率分别为93.7%～114%和90.5%～109%,血浆中检出限分别为30 μg·L-1和20μg·-1,肝脏中分别为25μg·L-1和15 μg·L-1,肾脏中均为10μg·L-1.结论:首次提出液相微萃取-非水后萃取方法,并将其成功应用于中药厚朴酚与和厚朴酚在大鼠体内的浓度测定.液相微萃取-非水后萃取能有效去除生物样品中干扰厚朴酚与和厚朴酚测定的内源性杂质,提高选择性.     

人血浆中西酞普兰的HPLC-荧光法测定

建立了HPLC-荧光法测定人血浆中西酞普兰的浓度.血浆样品经液-液萃取后测定,用Zorbax SB C8色谱柱,磷酸二氢铵缓冲液(pH 3.5)-乙腈(65:35)为流动相,激发波长240 nm,发射波长302 nm.血浆中西酞普兰线性范围为1～100ng/ml,最低定量浓度为1ng/ml,日内、日间RSD均小于4.0%,方法平均回收率为93.2%.     

反相高效液相色谱法测定狗血浆中番茄红素的浓度

目的建立用反相高效液相色谱法测定狗血浆中番茄红素浓度的方法.方法流动相为甲醇:乙腈:二氯甲烷(40:30:30, V/V),检测波长472 nm,柱温为室温,流速为1.0 mL·min-1.结果标准曲线的线性范围在0.012 4～0.496 μg·mL-1, r=0.999 2.平均萃取回收率为97.6%±4.2%,日内和日间的RSD分别在1.52%～4.95%、2.31%～7.38%之间.结论 所建立的方法灵敏、快速、重现性好、选择性强.     

Simultaneous determination of ranitidine and metronidazole in human plasma using high performance liquid chromatography with diode array detection

The development and validation of a simple method for the simultaneous determination of ranitidine and metronidazole in human plasma is described. Plasma samples (250 microL) were deproteinized by precipitation with 60% perchloric acid, centrifuged and the supernatant directly injected into the HPLC. Separation was achieved in isocratic mode with a Shimpak C(18) column and a mobile phase consisting of 10mM potassium dihydrogen phosphate pH 3.5:acetonitrile (90:10, v/v) with UV detection at 315 nm. The method showed good selectivity and sensitivity. Good and consistent recovery for metronidazole and ranitidine was obtained: 96.22+/-3.52 and 95.00+/-4.50% for ranitidine (25-1000 ng/mL) and metronidazole (60-10,000 ng/mL), respectively (n=3). With this one-step sample preparation method, both ranitidine and metronidazole could be quantified simultaneously in human plasma with good precision (R.S.D.<15%) and accuracy (bias values below 15%). The limit of quantification for ranitidine and metronidazole were 20 and 40 ng/mL plasma, respectively.     

Validated HPLC method for the determination of ranitidine in plasma

A validated HPLC method for the determination of ranitidine in human plasma is presented. Sulfanilamide as internal standard (IS) was used. Plasma samples were purified by solid phase extraction (SPE) using a copolymeric [poly(divinyl-benzene-co-N-vinylpyrrolidone)] column ("Oasis Waters"). Mobile phase consisting of dibasic potasium phosphate 0.08 M/acetonitrile/methanol/triethylamine 0.05% (89.5:3:7:0.05) pH5 was used at a flow rate of 0.9 ml/min on a C18 column (Nova-Pack, 3,9 x 300 mm, Waters). The eluate was monitored using an UVNis detector set at 300 nm. Ratio of peak area of ranitidine to sulfanilamide was used for the quantitation of plasma samples. FDA criteria for bioanalytical validation was used to validate the method. Linearity was assessed between 100-1600 ng/ml, the limit of quantitation was 100 ng/ml and recovery was greater than 94%. Accuracy, precision and selectivity met the current recommendations for bioanalytical method validation. The method was successfully used in a bioavailability study of a ranitidine tablet in healthy volunteers.     

Determination of ranitidine in plasma by high-performance liquid chromatography

A high-performance liquid chromatographic method has been developed for the determination of ranitidine in plasma. Ranitidine was extracted with acetonitrile by adding it to the plasma and then salting it out with potassium carbonate. The chromatographic column was 5-microns ODS silica, the mobile phase being acetonitrile-7 mM triethylammonium ion in phosphoric acid (pH 3.00) (30:70, v/v). The ranitidine peak was monitored at a wavelength of 315 nm, the retention time for ranitidine being 4.6 min. A limit of detection of 3 ng ml-1 was obtained for a 100-microliters injection of ranitidine. The method was found to be reproducible with a relative standard deviation (RSD) between 0.8-5.3% (n = 5) over the concentration range 25-80 ng ml-1 in plasma. The ranitidine concentration was determined in 18 different patients' plasmas. Ranitidine and its metabolites ranitidine S-oxide, ranitidine N-oxide and desmethyl-ranitidine, were also studied for chromatographic resolution from each other. It was shown that a group of common drugs did not interfere with ranitidine determination.     

The use of polymeric solid phase extraction and HPLC analysis for the determination of ranitidine in routine plasma samples obtained from paediatric patients

A sensitive HPLC method for the determination of ranitidine in small-volume (0.5 mL) paediatric plasma samples is described. Plasma samples were extracted using a simple, rapid solid phase extraction (SPE) technique developed using disposable copolymer packed SPE cartridges. Chromatographic separation was achieved by reverse-phase HPLC with isocratic elution using a microBondapak C18 column and a phosphate buffer (10 mM, pH 3.75)-acetonitrile (87:13 v/v) mobile phase with UV detection at 313 nm. The HPLC system exhibited linearity in the range 8-800 ng mL(-1). Intraday % CV and % bias values were in the range 1.28-8.09% (% bias -4.33 to -0.87) and interday % CV and % bias values were in the range 0.73-15.28% (% bias -1.80 to + 1.65). The limits of detection and quantitation obtained were 2 ng mL(-1) and 8 ng mL(-1), respectively, and ranitidine extraction recoveries from plasma ranged from 92.30 to 103.88%. In this study, the developed HPLC and SPE methodologies have been successfully applied to the determination of ranitidine concentrations in 68 paediatric plasma samples. The sampled population was drawn from patients already receiving the study drug therapeutically. Patients recruited had received ranitidine by two main routes - oral and intravenous. The plasma concentrations of ranitidine encountered in paediatric samples following oral or intravenous administration of a range of prescribed doses are presented graphically. These profiles are based on analysis of the first 68 plasma samples obtained from the first 35 patients recruited to the study receiving ranitidine by the oral or intravenous route.     

高效液相色谱法测定复方雷尼替丁胶囊中盐酸雷尼替丁的含量

目的建立测定复方雷尼替丁胶囊中盐酸雷尼替丁含量的方法。方法色谱柱：Kromasil C18（5μm,4.6mm×250 mm）;柱温：35℃;流动相：0.6%磷酸溶液（用50%氢氧化钠溶液调节pH值至7.10±0.05）-乙腈（83：17）;检测波长：230 nm;流速：1.0 mL min-1,进样量：10μL。结果盐酸雷尼替丁在0.010 85～0.54241 mg mL-1线性良好（r^2=0.999 7）,最小检出量为1.182 63 ng,平均回收率为99.8%（n=6）,RSD为1.0%。结论建立的方法简便、灵敏、专属性好、结果准确,适用于复方雷尼替丁胶囊中盐酸雷尼替丁含量的测定。     

Development of an HPLC method for the determination of ranitidine and cimetidine in human plasma following SPE

A selective, sensitive and accurate high-performance liquid chromatographic method has been developed, validated and applied for the determination of ranitidine and cimetidine in plasma samples. The effects of mobile phase composition, buffer concentration, mobile phase pH and concentration of organic modifiers on retention of investigated drugs were investigated. Sample preparation was carried out by adding an internal standard, famotidine, and the clean-up procedure was accomplished using solid-phase extraction (SPE). This method uses ultraviolet detection, the separation used a Lichrocart Lichrospher 60 RP-select B column and the mobile phase consisted of 0.2% triethylamine (TEA), 0.04 mol l(-1) KH2PO4 at pH 6.8 and 14% acetonitrile. The recovery, selectivity, linearity, precision and accuracy of the method were evaluated from spiked human plasma. The method has been implemented to monitor ranitidine levels in clinical samples.     

RP-HPLC法测定人血浆中富马酸喹硫平的浓度

目的:建立测定人体内富马酸喹硫平浓度的方法。方法:血样以重蒸乙醚处理后采用反相高效液相色谱法测定。色谱柱为Dikma Diamonsil C18,流动相为甲醇-5%三乙胺(50∶50),流速为1.0 ml/min,检测波长为250 nm。结果:富马酸喹硫平血药浓度在30¹ 500 ng/ml范围内线性关系良好,日内、日间RSD均<5%。结论:本试验所用方法线性关系良好,且具有专一性强、准确、灵敏度高的优点,符合生物样本分析的要求。可准确地测定人血浆样品中富马酸喹硫平的浓度。     

高效液相色谱法测定人血浆中阿立哌唑浓度

目的:建立以高效液相色谱法测定人血浆中阿立哌唑浓度的方法。方法:血浆样品经液-液提取后,采用高效液相色谱法检测。色谱柱为C18,流动相为0．03mol·L-1醋酸铵-乙腈(34:66),流速为0．8mL·min-1,柱温为40℃,检测波长为257nm,灵敏度为0．01 AUFS。结果:阿立哌唑检测浓度线性范围为5．0～600．0ng·mL-1(r=0．999 5),提取回收率均大于90％。结论:该方法灵敏、准确、快速,适用于人血浆中阿立哌唑浓度的检测。     

HPLC法测定人血浆中利福喷丁浓度

目的建立测定人血浆中利福喷丁的HPLC法。方法色谱柱为Kromasil C18（150mm×4．6mm,5μm）,流动相为甲醇：30mmol·L^-1 KH2PO4（磷酸调pH4．0）=69,5：30．5（v／v）．流速：1mL·min^-1,检测波长：335nm。结果利福喷丁在0．31-20μg·mL^-1。检测浓度范围内呈良好线型关系（r=0．9997）。高、中、低3种浓度的日内RSD〈5％,日间RSD〈10％,平均回收率分别为97．31％,103．52％．95．82％。结论该方法操作简便、灵敏、快速．适用于临床利福喷丁的血药浓度监测及其药动学研究。     

SPE-HPLC法检测病毒灭活血浆中的残留亚甲蓝

目的建立SPE-HPLC法检测病毒灭活血浆中亚甲蓝浓度的方法并对病毒灭活血浆中的残留亚甲蓝进行质控检测。方法血浆样品经Waters Oasis HLB固相萃取柱提取,采用迪马DiamonsialC18色谱柱(250mm×4.6mm,5μm),以甲醇-0.1%乙酸(45∶55,V/V)为流动相,流速为1.0mL·min-1,柱温30℃,检测波长651nm。结果亚甲蓝在0.05～5μmol.L-1范围内线性良好(r=0.9998),最低检出浓度为0.05μmol·L-1。日内、日间RSD均小于8%。结论SPE-HPLC法具有灵敏度高、特异性强的特点,用此法检测的病毒灭活血浆样品中的残留亚甲蓝均达到要求。