HPLC测定人血浆中西替利嗪的浓度

目的建立测定人血浆中西替利嗪浓度的高效液相色谱法。方法HypersilBDS-C18(150mm×5.0mm,5μm)色谱柱上,乙腈-0.05mol·L-1磷酸二氢铵(37∶70)为流动相,检测波长231nm,柱温30℃。结果西替利嗪在25～1.0×104μg·L-1范围内线性关系良好(r=0.9988),最低检测浓度为25μg·L-1。低、中、高3种浓度的回收率为99.4%～104.8%,日内、日间相对标准差为2.9%～9.6%。结论所用方法检测西替利嗪的血药浓度,结果准确可靠,可满足临床药物动力学研究的要求。     

HPLC法测定人血浆中西替利嗪的浓度

目的建立人血浆中西替利嗪浓度的HPLC测定法。方法色谱柱为Hypersil ODS C18(150mm×5.0mm,5μm);流动相为0.02mol.L-1磷酸二氢钾溶液(pH3.50)∶乙腈=77∶23;流速1.0mL.min-1;柱温30℃,检测波长230nm。结果西替利嗪血浆浓度在20~1000μg.L-1浓度范围内,线性关系良好(r=0.9998)。方法回收率99.54%~102.27%,日内、日间RSD均<10%。结论本方法简单、灵敏、重现性好,适用于临床血药浓度监测,以及人体药动学的研究。     

HPLC法同时测定盐酸西替利嗪和5种防腐剂的含量

目的 建立同时测定盐酸西替利嗪和5种防腐剂含量的方法。方法 采用高效液相色谱法,色谱柱为Inertsil ODS-3（4.6 mm×250 mm,5 μm）,流动相为乙腈:0.1%冰醋酸（40:60,V/V）,流速为1.5 mL?min^-1,检测波长为238 nm,柱温为30 ℃,进样量为20 μL。结果 盐酸西替利嗪和5种防腐剂的浓度与峰面积呈良好的线性关系（r≥0.999 7）;其精密度、稳定性试验的RSD＜2.0%;平均加样回收率分别为99.9%、99.8%、99.8%、99.7%、99.5%、100.2%,RSD分别为0.5%、0.5%、0.4%、0.5%、1.1%、1.2%。结论 本方法简便、高效、准确、重复性好,可用于盐酸西替利嗪和不同防腐剂的含量测定和质量控制。     

高效液相色谱法测定西替利嗪尿药浓度

目的：建立测定西替利嗪尿药浓度的反相离子对高效液相色谱法（ＲＰ－ＨＰＬＣ）。方法：以Ｎｏｖａ－ｐａｋＣ１８、４μｍ色谱柱为分离柱，流动相为乙腈∶磷酸盐缓冲液（０．０２ｍｏｌ·Ｌ－１、ＮａＨ２ＰＯ４）∶三乙胺（５０∶５０∶０．１５，Ｖ／Ｖ），ｐＨ值３．０７，内含十二烷基硫酸钠（ＳＤＳ）０．０１０５ｍｏｌ·Ｌ－１，去氯羟嗪为内标，ＵＶ检测波长２２９ｎｍ。结果：方法线性范围为０．２～２０μｇ·ｍｌ－１（ｒ＝０．９９９９），平均回收率为９６．４％±２．３％，日内和日间精密度均小于５％。结论：采用ＲＰ－ＨＰＬＣ法测定盐酸西替利嗪片的正常人尿药浓度，简便，迅速，准确     

Enantioselective determination of sibutramine and its active metabolites in human plasma

Although racemic sibutramine has been widely used for the treatment of obesity, its enantioselective detection method has not been elucidated in human plasma. In this report we introduce a validated analytical method for the determination of sibutramine and its two active metabolites, desmethylsibutramines using LC–MS/MS. R- and S-isomers of those compounds in human plasma were extracted using diethyl ether–hexane (4:1, v/v) followed by an addition of NaOH solution. After removing the organic layer, the residue was reconstituted in the mobile phase 10 mM ammonium acetate solution adjusted to pH 4.0 with acetic acid–acetonitrile (94:6, v/v). Both isomers in the extract were separated on a Chiralcel AGP chiral stationary-phase column and were quantified in a tandem mass spectrometry. The assay method was in accordance with FDA regulations for the validation of bioanalytical methods. This method was successfully used to profile the plasma concentrations of the stereoisomers of sibutramine and its two active metabolites with time in healthy volunteers.     

盐酸西替利嗪巴布膏剂的药动学和生物利用度研究

目的研究盐酸西替利嗪巴布膏剂在大鼠体内的药代动力学特征,并与口服给药的药动学进行比较,考察生物利用度。方法选用SD大鼠,分为巴布膏剂组和口服给药组,采用高效液相色谱法测定血药浓度;并剥取剩余的西替利嗪巴布膏剂,测定剩余药量。结果西替利嗪巴布膏剂组的T_(max)为2.35 h,C_(max)为2.68μg/mL,AUC为21.01μg·h/mL,MRT为23.26 h;西替利嗪口服组的T_(max)为1.11 h,C_(max)为11.65μg/mL,AUC为35.62μg·h/mL,MRT为12.13 h。扣除巴布膏剂中的剩余药量,按照实际进入皮肤的剂量计算得西替利嗪巴布膏剂相当于片剂的生物利用度为30.81%;不扣除巴布膏剂中的剩余药量,西替利嗪巴布膏剂相当于片剂的生物利用度为14.75%。结论西替利嗪巴布膏剂的血药浓度平稳而持久,具有缓释的特征。     

国产盐酸西替利嗪咀嚼片人体药动学研究

目的 研究国产盐酸西替利嗪咀嚼片在健康人体药动学.方法18名健康男性志愿者单剂量口服10 mg盐酸西替利嗪咀嚼片后取静脉血,采用LC-MS测定血浆中西替利嗪浓度,并用PKS药动学软件统计处理.结果 盐酸西替利嗪咀嚼片药-时曲线符合二室模型,其Cmax,Tmax,T1/2,AUC0-36,AUC0-∞分别为(25.21±24.35) μg·L-1,(1.6±0.3) h,(9.4±3.6) h,(1594.3±362.6) μg·L-1·h,(1688.0±498.6) μg·L-1·h.结论 本研究可为临床用药提供参考.     

高效液相色谱法测定人尿液中草酸的含量

目的:建立高效液相色谱(HPLC)法测定人尿草酸的含量。方法:采用Agilent SB C18(250mm×4.6mm,5μm)色谱柱;Agilent zorbax extend-C18(4.6mm×12.5mm,5μm)为预柱;以0.018mol.L-1磷酸二氢钾(含0.010mol.L-1四丁基硫酸氢铵和2.69×10-5mol.L-1EDTA,pH2.4~2.5)为流动相;检测波长210nm;流速1.5mL.min-1;柱温25℃;进样量20μL;以50g.L-1磺基水杨酸沉淀尿蛋白。结果:最低检测限为0.4mg.L-1。线性范围为3.125~100mg.L-1,平均回收率为97.35%,日内及日间精密度RSD<10.06%。结论:该方法简便,灵敏度高,重复性好,可用于尿路草酸钙结石成因的研究。     

HPLC测定人血浆中的盐酸西替利嗪

目的 建立测定人血浆中盐酸西替利嗪浓度的方法.方法 色谱柱为Hypersil BDS C18柱,流动相为乙腈-0.05 mol·L-1磷酸二氢铵溶液(37:70);流速1.5 ml·min-1;检测波长230 nm;盐酸羟嗪为内标.结果 线性范围为0.025～1.500 μg·ml-1,回归方程为Y＝2.482×10-2X 1.586×10-3(r＝0.9986),回收率97.7%,日内和日间RSD分别为5.5%、6.6%.结论 所建方法简便、准确、快速,适用于临床上盐酸西替利嗪药物动力学的研究.     

HPLC法测定人血浆中奥美拉唑的浓度

目的：了解人血浆中奥美拉唑浓度。方法：使用内标法,对10名健康志愿者的奥美拉唑血药浓度进行HPLC法检测。结果：分析结果显示,HPLC法的方法回收率为98.32%-104.96%,线性范围10~2000μg/L;10名志愿者的tmax=（2.09±0.75）h,t1/2=（1.02±0.58）h,ρmax=（774.56±340.46）μg/L,MRT0→1=（3.02±0.83）h,AUC0→12=（1751.42±1230.68）μg·h/L,AUC0→∞=（1784.50±1298.44）μg·h/L。结论：HPLC法测定奥美拉唑浓度,其灵敏度高,专属性强,准确,简便。     

高效液相色谱法测定人尿中盐酸多西环素

目的:建立测定人尿中多西环素含量的反相高效液相色谱方法。方法:以盐酸土霉素为内标,色谱分离采用Hyper-sil-ODS分析柱(4.6mm×250mm10μm),柱温30℃,以N,N-二甲基甲酰胺-0.05mol·L-1草酸铵缓冲液(40∶60,pH7.5)为流动相;流速:1.0mL·min-1,紫外检测波长:269nm。结果:尿中多西环素测定方法的线性范围为5~310mg·L-1,方法回收率平均为99.63%;日内精密度RSD小于0.49%、日间精密度RSD小于0.63%,最低检测限为0.03mg·L-1。多西环素尿样在检测过程中保持稳定。结论:本方法操作简便,灵敏度高,专一性强,可用于多西环素临床药动学研究。     

高效液相色谱法测定阿仑膦酸钠的尿药浓度

目的建立阿仑膦酸钠尿药浓度的高效液相色谱-荧光检测法。方法采用Shiseido Capcell Pak C18(150 mm×4.6 mm,5μm)色谱柱;梯度洗脱,流动相0.01 mol.L-1磷酸氢二钠(A)-乙腈(B);流速为1.0 mL.min-1;柱温35℃;内标物:帕米膦酸钠;荧光检测:激发波长为260 nm,发射波长为310 nm。结果阿仑膦酸钠峰与内标物峰面积比值在20～1 000μg.L-1呈良好的线性关系(r=0.996 1)。方法准确度与日内及日间RSD均符合要求,定量下限为20μg.L-1,提取回收率稳定,成功应用于4名男性健康受试者体内阿仑膦酸钠尿药浓度的测定。结论该法为临床研究阿仑膦酸钠体内尿药浓度提供了方法。     

RP-HPLC测定盐酸西替利嗪的含量及相关物质

目的　采用RP -HPLC法测定盐酸西替利嗪的含量及其有关物质。方法　采用ODS色谱柱 (5 μm ,4 .6mm× 15 0mm) ,流动相为乙腈 -水 -冰醋酸 (36∶6 4∶0 .1) ,流速 1.0ml·min-1,检测波长 2 2 9nm ,柱温为室温 (2 5℃ )。结果　盐酸西替利嗪测定的线性范围为 12 .5～ 15 0 μg·ml-1(r=0 .9998,n =6 ) ;日内RSD =0 .36 % (n =5 ) ,日间RSD =0 .6 4 % (n =5 ) ;最低检出限 0 .3ng。结论　所用方法简便、准确 ,专属性好     

高效液相色谱法测定人体血浆及尿液中多西环素的药物浓度

目的:建市人体血浆及尿液中多西环素药物浓度的高效液相色谱分析方法。方法:美他环素作为内标,待测血浆和尿液分别以高氯酸和甲醇沉淀蛋白,离心后取上清液进行高效液相色谱紫外检测,色谱柱为 Kromasil C_(18)柱(5μm,4.6mm×250mm);流动相为乙腈-水(30:70,含5 mmol·L~(-1)的柠檬酸),流速1.0 mL·min~(-1),检测波长350 nm。结果:本测定方法血浆和尿液的线性范围分别为O.1～6.13 mg·L~(-1)和0.085～13.62 mg·L~(-1),相关系数分别为0.9995和0.9991,血浆及尿液的相对回收率分别为95.2%～101.4%,95.6%～105.5%;日内日间 RSD 均小于10%,最低检测限分别为0.026 mg·L~(-1)及0.034mg·L~(-1)。结论:高效液相色谱法测定血浆与尿液中药物浓度简单、快速、可靠,可用于多西环素的药代动力学及生物利用度研究。     

13厂家盐酸西替利嗪片溶出度考察

目的比较国内不同厂家生产盐酸西替利嗪片溶出度的差异。方法采用转蓝法、紫外-可见分光光度法测定盐酸西替利嗪的含量,计算累积溶出量。结果其中11家企业的产品符合《中国药典》相关规定,2家生产企业2批次产品不符合《中国药典》相关规定。各厂家产品溶出行为差异较大。结论不同厂家生产的盐酸西替利嗪片的溶出行为存在较大差异,临床用药时应加以注意。     

HPLC法检查酒石酸布托啡诺原料药中右旋异构体

目的：采用HPLC法对酒石酸布托啡诺进行手性拆分并检测其中右旋异构体含量。方法：采用AstecCyclobondⅡ手性柱（β-CD polymer,250 mm×4.6 mm,5μm）,以2%的三乙胺醋酸盐溶液（pH 6.80）-甲醇（48∶52）为流动相。柱温：25℃,流速：1.0 mL/min,检测波长：280 nm。结果：本法对左、右旋酒石酸布托啡诺异构体的分离度为1.9,检测限为5 ng。结论：该方法操作简便,分离效果好,可用于酒石酸布托啡诺的质量控制。     

Development of chiral liquid chromatography-tandem mass spectrometry isotope dilution methods for the determination of unconjugated and total S-equol in human plasma and urine

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods for the determination of unconjugated and total (conjugated plus unconjugated) S-equol in human plasma and urine were developed and validated. The separation of R and S enantiomers was achieved with a Chiracel OJ-H column operated in a normal phase mode using ethanol/hexane mobile phase components. Ionization of S-equol by negative ion electrospray generated the [M-H](-) ion whose response was augmented by post-column addition of ammonium hydroxide. A triple stage quadrupole mass spectrometer was used to measure the ion current generated from the dissociative transitions m/z 241→m/z 121 (S-equol) and m/z 245→m/z 123 (equol-d(4)). The determination of total S-equol included an additional deconjugation step involving incubation of the sample with sulfatase and glucuronidase. Average recovery for both unconjugated and total S-equol was 85% with no observable matrix effects. Linearity was established for unconjugated S-equol from 0.025ng/mL to 10ng/mL (plasma) and 0.20ng/mL to 200ng/mL (urine). The average coefficient of variation and accuracy per occasion was within ±15% of the theoretical concentration of S-equol. The method was used to measure the pharmacokinetics of S-equol in human plasma after an oral administration of a single 20mg dose of S-equol to three normal healthy volunteers.     

Analysis of hesperetin enantiomers in human urine after ingestion of blood orange juice by using nano-liquid chromatography

Hesperetin (HT) is a flavanone abundantly found in citrus fruits. It has been reported that HT possesses significant antioxidant, anticancer, anti-inflammatory and analgesic activities. This explains the necessity of developing new methods more powerful and sensitive for analyzing HT in biological fluids. Taking into account the chiral nature of HT, the study of the stereospecific kinetics of in vitro and in vivo metabolism and tissue distribution could be a useful tool for further understanding stereoselective biotransformations in human body. A simple nano-liquid chromatographic method for the determination of the enantiomeric composition of hesperetin in human urine was developed. Chiral separation was achieved using a 100 μm I.D. capillary, packed with phenyl-carbamate-propyl-β-cyclodextrin stationary phase, employing a mobile phase composed by a mixture of triethylammonium acetate buffer (1%, v/v, pH 4.5) and water/methanol (30:70, v/v) at room temperature. The detection was done by using on-column UV detector at 205 nm. Calibration curves were linear in the studied concentration range from 0.25 to 25 μg/mL (r² > 0.999). Precision assay was <4.5% and was within 3% at the limit of quantification (0.5 μg/mL). The recovery of 7-ethoxycoumarin (IS), R- and S-hesperetin was greater than 82.48%, utilizing a liquid–liquid extraction procedure. The developed method was successfully applied to the determination of hesperetin enantiomers in urine samples obtained from a male volunteer, after the ingestion of 1 L of a commercial blood orange juice.