RP-HPLC-UV法测定大鼠血浆中西瑞香素浓度(英文)

建立了测定大鼠血浆中西瑞香素浓度的高效液相色谱法,并对其进行方法学确证。以蛋白沉淀法和液液萃取法对大鼠血浆中西瑞香素进行提取。采用Diamonsil C_(18)柱(200mm×4.6mm,5μm),流动相为甲醇-20mmol/L醋酸铵(醋酸调pH至3.2,42:58,v/v),流速1.0mL/min。检测波长345nm,柱温40℃。西瑞香素在0.020-2.00μg/mL浓度范围内线性关系良好(r>0.9900),定量下限0.020μg/mL。日内和日间精密度(RSD)为5.0%-10.6%,准确度(RE)为±(1.2%-2.5%)。所建立的方法成功应用于腹腔注射西瑞香素在大鼠体内的药代动力学研究。     

高效液相色谱法测定大鼠血浆中的西他列汀浓度及其药动学研究(英文)

本研究建立并验证了测定大鼠血浆中磷酸西他列汀浓度的高效液相色谱法,并在大鼠体内进行药动学研究。血浆中的药物和内标(氢化可的松)由乙酸乙酯萃取。所用色谱柱为Agilent Zorbax Extend-C18柱(250mm×4.6mm,4μm),进样量为20μL,检测波长为267nm,柱温为30oC。流动相为甲醇–水(60:40,内含10mMTris和10mM三乙胺,并用1M盐酸调至pH9.0)。流速为1.0mL/min。色谱峰具有良好的特异性。在0.75–100.0μg/mL范围内具有良好的线性关系(r>0.9957)。定量下限为0.75μg/mL。日间,日内精密度均小于10%。在0.75,10.0,100.0μg/mL的相对回收率为105.3%,99.8%,99.0%,提取回收率分别为81.5%,82.4%,84.5%。样品的稳定性良好。运用该方法测定了大鼠口服磷酸西他列汀水溶液后的血药浓度,说明本方法可用于药物动力学的研究。     

RP-HPLC法测定大鼠血浆中双嘧达莫浓度

目的建立大鼠血浆中双嘧达莫的RP-HPLC检测方法。方法采用RP-HPLC法测定大鼠血浆中药物浓度,色谱柱为Diamonsil ODS C18(200mm×4.6mm,5μm),流动相为甲醇-pH6.5磷酸二氢钾缓冲液(60∶40,v/v)。结果在0.10～10.0μg/mL范围内,双嘧达莫与内标峰面积比值与浓度线性关系良好(r=0.997),最低检测限为0.05μg/mL,绝对回收率为84.3%～92.3%,准确度为99.9%～103.1%,日内、日间RSD均<10%。结论本测定方法灵敏、准确、简便,适合于双嘧达莫的药代动力学研究。     

HPLC测定大鼠血浆中紫杉醇浓度

紫杉醇（paclitaxel）是近20年来发现的疗效显著的抗癌药物之一。与其他抗癌药物不同,紫杉醇是通过诱导和促进微管蛋白聚合、装配及稳定微管作用阻止肿瘤细胞生长,对卵巢癌、乳腺癌、头颈部癌、     

高效液相色谱法测定大鼠血浆中吴茱萸次碱浓度

建立大鼠血浆中吴茱萸次碱的反相高效液相色谱法。以乙腈 水 (5 8∶4 2 )为流动相 ,地西泮为内标 ,在波长 3 4 5nm处检测。吴茱萸次碱浓度在 0 0 2～ 1 6μg/mL呈线性 ,相关系数为 0 999,高中低 3种浓度平均萃取回收率均大于 80 0 %     

大鼠血浆中牡荆素鼠李糖苷的质量浓度测定

目的建立测定大鼠血浆中牡荆素鼠李糖苷质量浓度的反相高效液相色谱（RP-HPLC）法。方法以四氢呋喃-乙腈-0.5%冰醋酸（20：2.5：77.5）为流动相,血浆样品经简单的甲醇沉淀蛋白后,采用RP-HPLC法测定。结果血浆中牡荆素鼠李糖苷质量浓度的线性范围为0.2～80.0μg/mL,平均相对回收率为89.27%～98.25%,日内和日间精密度的RSD均小于10.79%,最低检测限为50ng/mL。结论建立的RP-HPLC法简便快速、灵敏度高,适用于牡荆素鼠李糖苷的药代动力学研究。     

LC-MS/MS快速测定大鼠血浆中HHY-002浓度

目的：建立用于测定治疗慢性心律失常药物HHY-002的大鼠血浆测定法。方法：流动相为乙腈-0．1％甲酸（65：35）,流速0．200mL／min,色谱柱Phenomenex LunaC18柱（150mm×2．00mm,5μm,5micron）,进样量为10μL,柱温35℃,以地西泮为内标。结果：HHY-002在1．98～1013．50ng／mL的范围内呈良好的线性关系（r=0．998）,最低定量限达1．98ng／mL,绝对回收率高于80％,日内和日间误差小于10％,方法回收率大于（81．4±4．5）％,符合生物样品分析要求。结论：建立的LC—MS／MS方法专属性强,灵敏度高,可用于HHY-002的体内定量分析。     

高效液相离子对色谱法测定糖尿病大鼠血浆中二甲双胍浓度及其药代动力学研究(英文)

本研究建立并验证了测定糖尿病大鼠血浆中二甲双胍浓度的高效液相色谱方法,以阿替洛尔为内标,加入10%高氯酸沉淀蛋白来制备血浆样品。色谱条件:AQ-C18极限色谱柱(250mm×4.6mm,5μm),流动相(pH5.05)为乙腈-水(31:69,v/v,水相中含有0.002M十二烷基磺酸钠,0.0125M磷酸二氢钾,0.015M三乙胺),流速1.0mL/min。二甲双胍在7.5-4000ng/mL范围内具有良好的线性关系(r>0.994),最低检测限(LLOQ)为7.5ng/mL,描述精密度的相对标准偏差(RSD)范围为1.87%-15.70%,准确度为93.98%-106.89%。二甲双胍和内标的回收率分别为95.40%和95.31%。不同实验条件下质控样品的稳定性相对偏差范围在±9.00%之内。该方法成功地运用到糖尿病大鼠单剂量灌胃10mg/kg二甲双胍后的药代动力学研究中,结果显示本研究采用AQ-C18极限色谱柱建立的离子对色谱法对强极性化合物如二甲双胍具有良好的分离效果。     

反相高效液相色谱法测定大鼠血浆中奥沙利铂的质量浓度

目的建立测定大鼠血浆中奥沙利铂质量浓度的反相高效液相色谱（RP-HPLC）法。方法色谱柱为Diamonsil^TMC₁₈柱（250 mm×4.6 mm,5μm）,流动相为乙腈-水（7:93）,检测波长为210 nm,流速为0.8 mL/min,柱温为30℃进样量为20μL。结果奥沙利铂保留时间为8.51 min,标准曲线方程为Y=8035.7X-11791（r=0.9995）,线性范围为2.0¹60.0μg/mL。低、中、高3个质量浓度奥沙利铂血浆样品的方法回收率分别为（112.7±3.61）%,（102.8±3.33）%,（101.6±4.60）%,RSD分别为3.20%,3.24%,4.53%;低、中、高3个质量浓度奥沙利铂血浆样品的提取回收率分别为（79.84±2.01）%,（82.29±3.28）%,（84.89±3.35）%,RSD分别为2.51%,3.98%,3.95%。血浆样品在-40℃反复冻融3次后性质稳定,对测定无影响。测定10只大鼠血浆样品中奥沙利铂的平均质量浓度为（67.4±10.45）μg/mL。结论该方法准确、简便、灵敏度高、重复性好,可为大鼠血浆中奥沙利铂的质量浓度测定提供方法学参考。     

HPLC法测定大鼠血浆中醋酸甲地孕酮的浓度

 目的：建立测定大鼠血浆中醋酸甲地孕酮浓度的HPLC法。方法：以达那唑为内标，使用正己烷进行萃取 ，采用HPLC法测定，色谱柱为Thermo BDS Hypersil C18柱（250 mm×4.6 mm，5 μm），柱温为30 ℃，流动相为甲醇- 水（75∶25），流速为1.0 mL•min-1，检测波长为288 nm。结果：在0.032～4 μg•mL-1范围内，醋酸甲地孕酮／达那 唑峰面积比（Y）与血浆中醋酸甲地孕酮的质量浓度（X）呈良好的线性关系，Y=270X+2.71×10-2，r=0.999 1（权重 因子1/c2）。方法的定量限为0.016 μg•mL-1，检测限低于0.006 4 μg•mL-1，醋酸甲地孕酮高、中、低3个浓度的日 内、日间精密度均小于15%，方法回收率为95%~105%，提取回收率为89%~95%。将建立的方法应用于实验室自制醋酸甲地 孕酮固体分散体在大鼠体内药动学性质研究中血药浓度的测定，证实了方法的可行性。结论：本方法准确可靠，可用于 大鼠血浆中醋酸甲地孕酮浓度的测定。     

HPLC测定大鼠血浆中的山奈酚

目的建立测定大鼠血浆中山奈酚的方法。方法以黄芩素为内标,无水乙醚为萃取溶剂,采用HPLC法,Shimm-pack VP-ODS柱为分析柱,乙腈-0.5%冰醋酸(33.3:66.7)为流动相,流速为1.0 ml.min-1,检测波长为370 nm,柱温为40℃。结果山奈酚0.050~5.301μg.ml-1与峰面积比(山奈酚/内标)的线性关系良好,最低定量限为0.020μg.ml-1,4种浓度的平均回收率为96.48%~108.33%,日内RSD≤7.39%,日间RSD≤5.88%,3种浓度的平均萃取回收率为93.67%、98.37%、101.83%。结论所建方法简单、灵敏、准确可靠,可用于山奈酚的药物动力学研究。     

Quantitative determination of daumone in rat plasma by liquid chromatography-mass spectrometry

Daumone, 6-(3,5-dihydroxy-6-methyl-tetrahydro-pyran-2-yloxy)-heptanoic acid is a pheromone secreted by Caenorhabditis elegans, and has been known as a pivotal regulator of chemosensory processes in development and ageing. A quantification method using mass spectrometry was developed for the determination of daumone in rat plasma. After simple protein precipitation with acetonitrile including an internal standard, the analytes were chromatographed on a reversed-phase column and detected by liquid chromatography/tandem mass spectrometry with electrospray ionization. The accuracy and precision of the assay were in accordance with FDA regulations for validation of bioanalytical methods. This method was applied to measure the plasma daumone concentrations after a single intravenous administration of daumone in rats.     

HPLC-electrospray mass spectrometric assay for the determination of (R,R)-fenoterol in rat plasma

A fast and specific liquid chromatography-mass spectrometry method for the determination of (R,R)-fenoterol ((R,R)-Fen) in rat plasma has been developed and validated. (R,R)-Fen was extracted from 125 microl of plasma using solid phase extraction and analyzed on Atlantis HILIC Silica 3 microm column. The mobile phase was composed of acetonitrile:ammonium acetate (pH 4.1; 20mM) (85:15, v/v), at a flow rate of 0.2 ml/min. The lower limit of detection (LLOD) was 2 ng/ml . The procedure was validated and applied to the analysis of plasma samples from rats previously administered (R,R)-Fen in an intravenous bolus.     

固相萃取用于血浆中抗癫痫药物的提取

目的：探讨固相萃取法用于血浆中抗癫痫药物的提取。方法：以活化的ＯＤＳ柱（４０ｕｍ）为固相萃取柱,二氯甲冠为洗脱剂,洗脱液浓缩后以ＨＰＬＣ法测定药物的浓度。结果：苯巴比妥、苯妥英在５．００～６０．００ｕｇ／ｍｌ范围内,卡马西平在２．５０～２０．００ｕｇ／ｍｌ范围内线性关系良好,ｒ分别为０．９９８０,０．９９９９,０．９９９０;绝对回收率分别为５９．６０％～６８．７％,５８．９％～６８．０％,５３．８     

Pharmacokinetic study of six flavones in rat plasma and tissues after oral administration of 'JiangYaBiFeng' using SPE-HPLC-DAD

In this study, a high performance liquid chromatography (HPLC) coupled with diode array detection (DAD) for simultaneous determination of six flavones including baicalein, sophoricoside, rutin, baicalin, quercetin and genistein in rat plasma and tissues after oral administration of JiangYaBifeng (JYBF) tablets was developed. The investigated analytes in plasma and tissues were extracted and purified with liquid-liquid extraction and solid phase extraction (SPE). Chromatographic separation was accomplished on a DIONEX Acclaim C18 column (250 mm × 4.6 mm, 5.0 μm particle size) with a simple linear gradient elution. The calibration curves for all the flavones had good linearity in the measured range with R² higher than 0.9983. The relative errors (REs) of the intra- and inter-day accuracy at different flavones levels were all less than ±10%. The proposed method enables unambiguous identification and quantification of investigated flavones in vivo. This is the first report on determination of the major flavones in rat plasma and tissues after oral administration of JYBF tablets. The results provided a meaningful basis for evaluating the clinical application of this medicine.     

Sensitive determination of a pharmaceutical compound and its metabolites in human plasma by ultra-high performance liquid chromatography-tandem mass spectrometry with on-line solid-phase extraction

This paper describes the determination of a drug candidate and two metabolites in human plasma by column-switching LC-MS/MS after protein precipitation. Starting from a standard method with a quantitation limit of 0.5 ng/mL, a highly sensitive assay was developed, employing UHPLC separation and detection on an API 5000 mass spectrometer. The injected plasma equivalent was increased from 6 to 20 μL; conventional column trapping for compound enrichment and removal of matrix constituents was combined with high-pressure analytical separation using small particle columns to improve resolution and signal-to-noise ratio. Quantitation limits were thus lowered to between 5 and 20 pg/mL, offering the possibility to provide bioanalytical support for microdosing studies in humans. Excellent assay quality and robustness were achieved by both methods.     

固相萃取用于血浆中抗癫痫药物的提取

目的：探讨固相萃取法用于血浆中抗癫痫药物的提取。方法：以活化的ＯＤＳ 柱（４０μｍ） 为固相萃取柱,二氯甲烷为洗脱剂,洗脱液浓缩后以ＨＰＬＣ 法测定药物的浓度。结果：苯巴比妥、苯妥英在５ ．００ ～６０ ．００μｇｍｌ 范围内,卡马西平在２ ．５０ ～２０ ．００μｇｍｌ 范围内线性关系良好,ｒ 分别为０ ．９９８０ ,０ ．９９９９ ,０ ．９９９０ ;绝对回收率分别为５９ ．６０ ％～６８ ．７ ％ ,５８ ．９ ％ ～６８ ．０ ％ ,５３ ．８ ％ ～５７ ．３ ％ ;相对回收率分别为９２ ．２ ％ ～９８ ．６ ％ ,９７ ．１ ％ ～１０２ ．６ ％ ,９７ ．９ ％ ～１０２ ．０ ％ ,ＲＳＤ＜ ６ ％ ;血浓监测结果与经传统的液液萃取后所得结果非常接近。结论：固相萃取法操作简单、省时、提取干净,可用于血浆中抗癫阅药物的提取。     

Quantitative assay for bradykinin in rat plasma by liquid chromatography coupled to tandem mass spectrometry

An assay to quantify bradykinin in rat plasma has been developed and validated, using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Sar-D-Phe(8)-des-Arg(9)-bradykinin was used as internal standard. Aprotinin was added to rat plasma to inhibit the activity of proteinases. Recoveries for solid-phase extraction (SPE) on Strata X reversed phase were greater than 80%. Multiple reaction monitoring (MRM) on a triple quadrupole mass spectrometer equipped with an electrospray source (ESI), operating in the positive ion-mode, was used for detection. The assay was validated and stability was explored. Bradykinin (10-500 ng/mL) was quantified with accuracy values (% RE) below 10% and intra- and inter-day precisions (% RSD) below 12 and 16%, respectively, for all concentrations. The method was successfully applied to several plasma samples from low levels kallikrein rats (LKRs) compared with normal kallikrein rats (NKRs).     

Quantitative determination of spinosin in rat plasma by liquid chromatography-tandem mass spectrometry method

A sensitive method for the quantitative determination of spinosin in rat plasma was developed and validated using high-performance liquid chromatographic separation with tandem mass spectrometric detection. The analytes of interest were extracted from rat plasma samples by methyl tert-butyl ether (MTBE) after acidification with 1.0% acetic acid aqueous solution. Chromatographic separation was achieved on an Agilent Zorbax SB-C(18) (50 mm x 4.6 mm, 5 microm) using a isocratic mobile phase consisting of acetonitrile-water (30:70, v/v) with 1% isopropyl alcohol and 0.01% heptafluorobutyric acid. The flow rate was 0.2 ml/min. The column temperature was maintained at 25 degrees C. Detection was performed on a triple quadrupole tandem mass spectrometer by selected reaction monitoring (SRM) mode via electrospray ionization (ESI). The calibration curve was linear over the range of 1.00-400 ng/ml in rat plasma, with 1.00 ng/ml of the lower limit of quantification (LLOQ). The inter- and intra-day precisions and accuracy for all samples were satisfactory. The validated method was successfully applied for the pharmacokinetic study of spinosin in rat. After oral administration of 20mg/kg spinosin to rats, the main pharmacokinetic parameters of T(max), C(max), T(0.5) and AUC(0-T) were 5.33+/-0.58 h, 132.2+/-10.6 ng/ml, 4.89+/-0.37 h, 1.02+/-0.09 microg h/l, respectively.