微透析联用反相高效液相色谱对大鼠皮肤葛根素的药代动力学研究

采用在体微透析取样技术联用RP-HPLC的方法对大鼠皮肤中的葛根素进行了体内药代动力学的考察.将微透析探针进行体内外校正后,植入大鼠皮下,以磷酸盐缓冲溶液作为灌流液,收集的样品在以Hypersil-C18(150 mm×4.6 mm, 5 μm)为色谱柱,流动相为V(甲醇)∶ V(0.1% 醋酸)＝30∶ 70,流速1 mL/min,检测波长221 nm的HPLC色谱条件下进行分析,所得的大鼠皮下游离葛根素浓度用Kinetica软件进行处理.结果显示:微透析探针体内外校正结果显示葛根素适宜应用微透析技术,在所建立的HPLC条件下葛根素在0.1～100 mg/L范围内线性关系良好(r=0.9999, n=6),浓度为100、10和0.1 mg/L的葛根素日内RSD分别为0.38%、0.80%和1.21%(n=6);日间RSD分别为0.44%、0.73%和1.68%(n=6),所得数据经Kinetica软件拟合后,得到大鼠皮下非结合葛根素药-时曲线和主要药动学参数.所建立的微透析联用RP-HPLC方法能够在体、实时监测大鼠皮下葛根素浓度.     

反相高效液相色谱法测定兔血清中拉莫三嗪的浓度及其动物药代动力学研究

 用反相高效液相色谱法测定了兔血清中抗癫痫药拉莫三嗪的浓度 ,以苯乙酮作内标 ,甲醇 磷酸二氢钾 ( 0 0 5mol/L ,pH 3 8)为流动相 ,经YWG ODS柱 ( 2 0 0mm× 3 6mmi d )分离 ,波长为 3 0 5nm的二极管阵列检测器检测 ,结果血药浓度在 0 2 5mg/L～ 5 0mg/L范围内线性良好 (r =0 9998) ,平均回收率为 93 78% ,标准误差为2 66% ,日内和日间RSD分别为 2 4 % (n =9)和 5 3 % (n =5 )。取家兔经胃给药 ( 2 5mg/kg) ,在设定时间抽血 ,测定动物药代动力学参数 ,经 3P87药代动力学程序包拟合 ,结果为Cmax=9 3 4 μg/mL ,t1/ 2 (Ke) =8 78h。     

Competing binding of metal ions with protein studied by microdialysis

A method has been established to study the competing binding of metal ions with protein by a combined technique of microdialysis with high performance liquid chromatography (HPLC). Ni2+, Cd2+, Zn2+, Cu2+ and human serum albumin (HSA) were chosen as model metal ions and protein. The experimental results show that Ni2+ and Cu2+ share a common primary binding site on HSA, and Zn2+ and Cd2+ share a different common primary binding site from them, but there is a common multi-metal binding site for all of those four metal ions. This method show advantages of fast sampling, easily to be operated and especially to be useful when ideal spectroscopic probes are not available for the study of interaction between protein and metal ions.     

Pharmacokinetics of monoester‐diterpenoid alkaloids in myocardial infarction and normal rats after oral administration of Sini decoction by microdialysis combined with liquid chromatography–tandem mass spectrometry

Monoester‐diterpenoid alkaloids are the main bioactive components of Sini decoction, which is a well‐known traditional Chinese medicine formula for the treatment of myocardial infarction (MI) and heart failure in China. In this work, an ultra‐high‐performance liquid chromatography–mass spectrometry combined with microdialysis method was successfully established and applied for investigating for the first time comparative plasma pharmacokinetics of three monoester‐diterpenoid alkaloids (benzoylmesaconitine, benzoylaconitine and benzoylhypacoitine) in normal and MI rats after oral administration of Sini decoction. The statistical results of pharmacokinetic parameters demonstrated that benzoylmesaconitine, benzoylaconitine and benzoylhypacoitine showed lower peak concentration, longer half‐life, smaller area under the concentration–time curve, slower clearance, time to peak concentration and mean residence time in MI rats than in normal rats (p < 0.05), which indicated that monoester‐diterpenoid alkaloids exhibited lower systemic exposure and slower elimination in the MI rats. The results provided the experimental basis for understanding the metabolic fate and therapeutic effects of Sini decoction.     

大黄蒽醌衍生物的高效液相色谱法测定及在家兔体内的药代动力学研究

建立了家兔血浆中大黄蒽醌衍生物的高效液相色谱测定方法,并用于含大黄单 复方制剂中蒽醌衍生物在家兔体内的药代动力学研究。血样经处理后,在μ-Bondapak C 18柱上分析,流动相为甲醇-乙腈-0.2 mol/L磷酸二氢钠溶液(体积比为27∶35.5∶37.5),磷酸调pH 2.8,检测波长为225 nm。在此色谱条件下,各蒽醌衍生物与内标1,8-二羟基蒽醌的分离度符合要求,血浆内源性成分无干扰。两种制剂灌胃后的血药浓度-时间曲线符合二室模型, 二者的AUC无显著性差异。该法灵敏 准确,适用于含大黄的     

Determination of picroside II in dog plasma by HPLC and its application in a pharmacokinetics study

A sensitive and simple high-performance liquid chromatography method with UV detection was developed and validated for determining picroside II in dog plasma. Paeoniflorin was employed as internal standard and the sample pre-treatment procedure consists of deproteinization by addition of acetonitrile. Chromatographic separations were performed on a Shimadzu VP-ODS column (250 x 4.6 mm i.d., 5 microm). The mobile phase consisted of acetonitrile-0.1% acetic acid aqueous (v/v), 23:77, v/v, at a rate of 1 mL/min. Detection was carried out at a wavelength of 266 nm. Calibration standards ranged from 0.25 to 500 microg/mL in dog plasma and the mean correlation coefficient of 0.9981 was found for the linear calibration curves (n = 6). The limit of quantification (LOQ) was 0.25 microg/mL. Intra- and inter-assay RSD ranged from 0.70 to 7.5%. Accuracy (%bias) ranged from -6.3 to 6.0%. This method was applied to the pharmacokinetic study of picroside II in dogs. The study demonstrated the plasma picroside II concentration-time curves were fitted to the two-compartment open model and showed linear pharmacokinetics.     

Simultaneous detection of four flavonoids and two alkaloids in rat plasma by LC–MS/MS and its application to a comparative study of the pharmacokinetics between Abri Herba and Abri mollis Herba extract after oral administration

Abri Herba and Abri mollis Herba both were important members of the Leguminosae family in southwestern China. Abri mollis Herba was often used as Abri Herba due to their proximity, but there are few studies on pharmacokinetics to compare their main identical active compositions. A sensitive and selective high‐performance liquid chromatography with tandem mass spectrometry method in the positive/negative electrospray ionization switching mode was developed and validated for the simultaneous analysis of four flavonoids and two alkaloids in rat plasma. The chromatographic separation was carried out on a C18 column with a gradient mobile phase consisting of methanol and 0.5% acetic acid. The detection of the target compounds was conducted in multiple‐reaction monitoring mode with a hybrid triple quadrupole linear ion trap mass spectrometer equipped with positive/negative ion‐switching electrospray ion source. The differences in pharmacokinetics were discovered, which indicated that the substitution between them is some degree of irrationality. The validated method was successfully applied to pharmacokinetic study of the six components in male rat plasma after oral administration of Abri Herba and Abri mollis Herba extract and the results in the study would provide a useful guide for the clinical application of Abri Herba with those in Abri mollis Herba.     

Determination of GL‐V9, a derivative of wogonin,in rat plasma by UPLC–MS/MS and its application to a pharmacokinetic study after oral and pulmonary administration

GL‐V9, a derivative of wogonin, shows much more potent anticancer properties than wogonin. In this study, a selective, sensitive and rapid ultra‐high‐performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) method was developed and validated for the determination of GL‐V9 in rat plasma. Plasma samples were processed using methanol to precipitate protein. Chromatographic separation of analytes was achieved on a C₁₈ column using gradient elution within 4.5 min. The mobile phase consisted of acetonitrile and water including 0.1% (v/v) formic acid and 5 mm ammonium acetate. GL‐V9 and caffeine (internal standard) were monitored by positive electrospray triple quadrupole mass spectrometer and quantified using multiple reaction monitoring (MRM) mode with the transitions of m/z 410.20 → 126.10 (GL‐V9) and 195.10 → 138.00 (IS: caffeine), respectively. Good linearity was obtained over the range of 2–1000 ng/mL (R² > 0.99) and the extraction recovery was 101.91 ± 11.34%. The intra‐ and inter‐day precision variations were small (RSD 1.35–6.96%) and the relative error (RE) of accuracy was ?7.35–6.27%. The established and validated UPLC–MS/MS method was successfully applied to study the pharmacokinetic behavior of GL‐V9 after administration through different delivery routes. The results demonstrated that pulmonary delivery exhibited a greater advantage in terms of improving bioavailability compared with oral administration.     

Determination of ferruginol in rat plasma via high‐performance liquid chromatography and its application in pharmacokinetics study

Ferruginol, a diterpene phenol, has recently received attention for its extensive pharmacological properties, including anti‐tumor, antibacterial, cardio‐protective and gastroprotective effects. In the present study, a high‐performance liquid chromatographic (HPLC) method was developed for determination of ferruginol in rat plasma and applied for the pharmacokinetics study. The HPLC assay was performed with a VP ODS‐C₁₈ column. The mobile phaseconsisted of methanol and 1% acetic acid solution (90:10, v/v). The flow rate was 1.0 mL/min, and the wavelength was set at 270 nm. This method was linear over the studied range of 0.1–10.0 µg/mL for ferruginol. The correlation coefficient was 0.9998. The intra‐day and inter‐day precisions were better than 4 and 5%, respectively. The extraction recovery and accuracy were greater than 97 and 96%, respectively. The detection limit was 30 ng/mL. The mean maximum concentration of ferruginol in rat plasma was 3.14 µg/mL at 40 min after oral administration at a dose of 20 mg/kg. Ferruginol was absorbed quickly p.o. with t_1/2ka = 14.86 min and had a high rate of elimination with t_1/2 = 41.73 min. The pharmacokinetic process of ferruginol in rat was well described with a one‐compartment model. Copyright © 2009 John Wiley & Sons, Ltd.     

Determination of tectoridin in rat plasma by high‐performance liquid chromatography and its application to pharmacokinetic studies

A simple, specific and sensitive HPLC method with UV detection was developed and validated for the determination of tectoridin in rat plasma for the first time. Chromatographic separation was performed on a Welchrom^TM C₁₈ column (150 × 4.6 mm, i.d., 5 µm) at a flow rate of 1.0 mL min^?1, using a mixture of methanol–2% HAc aqueous solution (31:69, v/v) as the mobile phase with UV detection at 266 nm. The calibration curves for tectoridin were linear over the concentration range of 1.10–274.40 µg mL^?1 in rat plasma. The intra‐ and inter‐day accuracies (RE) were within ?3.23% and 4.11%. The intra‐ and inter‐day precisions (RSD) were not more than 2.74 and 4.72%, respectively. The present method was successfully applied to the pharmacokinetic studies of tectoridin in rats after intravenous administration of three different doses. Copyright © 2009 John Wiley & Sons, Ltd.     

Pharmacokinetics and metabolism of olerciamide A from Portulaca oleracea L. in rats by UHPLC‐UV and UHPLC‐ESI‐Q‐TOF/MS

The aim of this study was to elucidate the pharmacokinetics of olerciamide A in rats after oral and intravenous administration of Portulaca oleracea L. extract by a simple and rapid ultra high‐performance liquid chromatography method with bergapten as internal standard. The pharmacokinetic results indicated that olerciamide A was rapidly distributed with a time to peak concentration of 30 min after oral administration and presented a low oral absolute bioavailability of 4.57%. The metabolism of olerciamide A in rats was also investigated using ultra‐high‐performance liquid chromatography electrospray coupled with quadrupole–time of flight mass spectrometry to elucidate the reason for the low absolute bioavailability of olerciamide A and seven metabolites of oleraciamide A were found in rat plasma and urine.     

Pharmacokinetics of 13 active components in a rat model of middle cerebral artery occlusion after intravenous injection of Radix Salviae miltiorrhizae‐Lignum dalbergiae odoriferae prescription

As a representative formulation of Radix Salviae miltiorrhizae (Danshen)‐Lignum Dalbergiae odoriferae (Jiangxiang), Xiangdan injection is widely prescribed for cardio‐ and cerebrovascular diseases in practice. This necessitates a pharmacokinetic investigation of this formulation to make it safer and more broadly applicable. We developed and validated a sensitive, selective, and reliable high‐performance liquid chromatography with tandem mass spectrometry method for the simultaneous determination of 11 phenolic compounds including danshensu plus two diterpenoid quinones like cryptotanshinone and tanshinone IIA in rat. We applied this method for the pharmacokinetic studies of the 13 compounds in a rat model of middle cerebral artery occlusion after intravenous injection of Xiangdan injection or Danshen injection. In sham‐operated rats, the animals taking Xiangdan injection exhibited significant growth of the area under the curve for danshensu, protocatechuic aldehyde, and tanshinone IIA compared with the changes seen in the data of those administrated with Danshen injection. Such a pattern was also observed in middle cerebral artery occlusion rats, whereas increased the area under the curve values were observed for danshensu, protocatechuic aldehyde, caffeic acid, rosmarinic acid, and tanshinone IIA. These results demonstrated that synergistic interactions occurred between the components of Danshen and the active compounds of Jiangxiang both in sham‐operated and middle cerebral artery occlusion rats, increasing the bioavailability of Danshen. The results presented herein can be used to determine a reference dose for the clinical application of Xiangdan injection, and to elucidate the synergistic mechanism of Danshen and Jiangxiang.     

高效液相色谱-电化学检测法测定鼠脑微透析液中谷胱甘肽和半胱氨酸

采用高效液相色谱法,用电化学检测手段成功地测定了沙土鼠脑纹状体微透析液内的谷胱甘肽（ＧＳＨ）和半胱氨酸（Ｃｙｓ）。检测前用电化学自清洗方式能有效地提高玻璃碳电极的检测水平,使其具有较好的重现性和回收率。Ｃｙｓ和ＧＳＨ的回收率分别为９１．４％和８７．３％。电化学检测的工作电压为０．９Ｖ,用外标法定量。由微透析探针获取的纹状体透析液Ｃｙｓ和ＧＳＨ的浓度分别为６８．５９和６５．５４ｎｍｏｌ／Ｌ,验证了方法的实用性和可靠性。     

反相高效液相色谱法测定人血浆中曲尼司特浓度

报道了用反相高效液相色谱法测定人血浆中曲尼司特浓度。以甲醇－磷酸二氢钾缓冲液（６０∶４０，Ｖ／Ｖ；ｐＨ４．２）为流动相，色谱柱采用ＹＷＧ－Ｃ１８不锈钢柱，紫外３３３ｎｍ波长检测，血浆样品经甲醇沉淀蛋白后直接进样测定。血浆浓度在０．６５～４０ｍｇ／Ｌ范围内线性良好，方法的平均回收率为１００．０％±４．１％。血浆最低检测浓度为０．２ｍｇ／Ｌ。对１０名健康志愿者口服２００ｍｇ曲尼司特后不同时间的血药浓度进行了测定。     

Simultaneous determination and pharmacokinetics study of four quinones in rat plasma by ultra high performance liquid chromatography with electrospray ionization tandem mass spectrometry after the oral administration of Qianzhi capsules

A sensitive, specific, and accurate ultra high‐performance liquid chromatography with electrospray ionization tandem mass spectrometry method was developed and validated for the simultaneous quantification of purpurin, munjistin, mollugin, and alizarin from Qianzhi capsules in rat plasma. Chromatographic separation was performed on an Agilent Eclipse Plus C₁₈ RRHD column with a mobile phase consisting of methanol and 5 mM ammonium acetate/water with gradient elution. The analytes were quantified on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring mode and switching the electrospray ion source polarity with positive electrospray ionization in a single run. Samples were pretreated by liquid–liquid extraction with cyclohexane. The intra‐ and interday precision and accuracy of the assay were within acceptable ranges. Matrix effects for all of the analytes were between 90.16 and 100.21%. The average recovery ranged from 75.38 to 88.96%. This method was successfully applied to study the pharmacokinetic parameters of the four compounds in rat plasma after oral administration of Qianzhi capsules. Four quinones could be rapidly absorbed into blood (t_max, 0.80–1.93 h) and eliminated relatively slowly (t_1/2, 8.07–11.97 h). The results might be helpful for guiding the clinical application of Qianzhi capsules in the future.     

高效液相色谱法测定人血清中头孢克罗

建立了测定人血清中头孢克罗的高效液相色谱法。分析柱为Ｓｈｉｍ－ｐａｃｋＣＬＣ－ＯＤＳ（５μｍ）,６．０ｍｍ×１５０ｍｍ;流动相为磷酸盐缓冲液（ｐＨ５．８）∶乙腈＝８７．５∶１２．５,流速为１．２ｍＬ／ｍｉｎ;紫外检测波长为２５４ｎｍ。血清中头孢克罗浓度在０．１２５～２４ｍｇ／Ｌ范围内,峰高与浓度呈良好的线性关系,回归方程为Ｈ＝－３．２００＋９２．２９Ｃ（ｎ＝９,ｒ＝０．９９９９）,日内、日间变异系数分别为４．１％和５．８％,平均回收率为９９．３％。     

除草剂西玛津与过氧化氢酶的相互作用

应用荧光光谱法和微透析液相色谱法研究了水溶液中除草剂西玛津与过氧化氢酶分子间的相互作用。结果表明 ,除草剂对过氧化氢酶的荧光有较强的猝灭作用 ,且静态猝灭是引起荧光猝灭的主要原因。从荧光猝灭和以Scatchard方程拟合微透析液相色谱法的测定结果求出除草剂和CAT的结合常数及结合位点数分别为K =1.5 5× 10 4 L/mol,n =0 .94。并依据能量转移机制 ,求出了西玛津和CAT相互结合时 ,给体 受体间距离r为 0 .164nm。西玛津与过氧化氢酶的相互结合作用以静态猝灭过程为主 ,且其猝灭机制是通过能量转移产生的。西玛津可能与CAT的Tyr2 14发生结合作用     

Validated HPLC method for the quantitative determination of CoQ(10) in dog plasma and its application to a pharmacokinetic study

Coenzyme Q₁₀ (CoQ₁₀) is a naturally occurring compound located in all membranes throughout the cell. A rapid and sensitive HPLC method was developed to determine the concentration of CoQ₁₀ in dog plasma using a surrogate matrix. Chromatographic separation was carried out on a Diamonsil C₁₈ column with the UV detector set at 275 nm. Methanol–2‐propanol (40:60, v/v) was used as a mobile phase delivered at a flow rate of 1.0 mL/min. Calibrators were prepared using blank plasma–K₂HPO₄ buffer (50 mm , pH 8.0)–saline (1:3:6, v/v/v) as surrogate matrix. It was shown that the surrogate matrix had similar properties to dog plasma for CoQ₁₀ in extraction, freeze–thaw and stability. The assay was linear over the concentration range of 0.10–100 µg/mL. The intra‐ and inter‐day precisions were within 13.3% in terms of relative standard deviation (RSD%) and the accuracy was within ±7.5% in terms of relative error. This simple and reproducible HPLC method with less plasma volume (0.4 mL) and adequate sensitivity was successfully applied to pharmacokinetic studies of CoQ₁₀ in dogs and an investigation of the effect of CoQ₁₀ formulation on CoQ₁₀ baseline levels. Copyright © 2010 John Wiley & Sons, Ltd.     

Quantification of Levosulpiride in Human Plasma by High‐Performance Liquid Chromatography

Abstract

An analytical procedure was developed and validated for the quantification of levosulpiride in human plasma. After subjecting a plasma sample to a two‐step extraction procedure, an aliquot of the aqueous phase was injected onto an high‐performance liquid chromatography system equipped with a fluorescence detector. The detector response was linear for levosulpiride concentrations in the range of 2.5 to 500 ng/ml. The intra‐ and inter‐day precision was below 15.4 and 10.1%, and the accuracy was in a range from 89.7 to 109.4%. The method is applicable for use in the pharmacokinetic characterization of levosulpiride after a 75‐mg oral dose in humans.     

PEEK tube‐based online solid‐phase microextraction–high‐performance liquid chromatography for the determination of yohimbine in rat plasma and its application in pharmacokinetics study

Yohimbine is a novel compound for the treatment of erectile dysfunction derived from natural products, and pharmacokinetic study is important for its further development as a new medicine. In this work, we developed a novel PEEK tube‐based solid‐phase microextraction (SPME)–HPLC method for analysis of yohimbine in plasma and further for pharmacokinetic study. Poly (AA‐EGDMA) was synthesized inside a PEEK tube as the sorbent for microextraction of yohimbine, and parameters that could influence extraction efficiency were systematically investigated. Under optimum conditions, the PEEK tube‐based SPME method exhibits excellent enrichment efficiency towards yohimbine. By using berberine as internal standard, an online SPME‐HPLC method was developed for analysis of yohimbine in human plasma sample. The method has wide linear range (2–1000 ng/mL) with an R ² of 0.9962; the limit of detection was determined and was as low as 0.1 ng/mL using UV detection. Finally, a pharmacokinetic study of yohimbine was carried out by the online SPME‐HPLC method and the results have been compared with those of reported methods.