Development and validation of amisulpride in human plasma by HPLC coupled with tandem mass spectrometry and its application to a pharmacokinetic study

In this study, authors developed a simple, sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantification of Amisulpride in human plasma using Amisulpride-d(5) as an internal standard (IS). Chromatographic separation was performed on Zorbax Bonus-RP C18, 4.6 × 75 mm, 3.5 μm column with an isocratic mobile phase composed of 0.2% formic acid:methanol (35:65 v/v), at a flow-rate of 0.5 mL/min. Amisulpride, Amisulpride-d(5) was detected at m/z 370.1→242.1 and 375.1→242.1. The drug and the IS were extracted by a liquid-liquid extraction method. The method was validated over a linear concentration range of 2.0-2500.0 ng/mL for Amisulpride with a correlation coefficient of (r(2)) ≥ 0.9982. This method demonstrated intra- and inter-day precision within 0.9 to 1.7 and 1.5 to 2.8 % and intra- and inter-day accuracy within 98.3 to 101.5 and 96.0 to 101.0 % for Amisulpride. Amisulpride was found to be stable at 3 freeze-thaw cycles, bench top and auto sampler stability studies. The developed method was successfully applied to a pharmacokinetic study.     

HPLC法测人血浆对乙酰氨基酚浓度及其药代动力学研究

目的建立HPLC法测人血浆中对乙酰氨基酚浓度,并进行药代动力学研究.方法血浆样品采用液-液萃取法萃取,UltimateTM XB-C18(250mm×4.6mm,5μm)色谱柱分离;流动相为乙腈:水(10∶90,V/V),流速:1.0mL·min-1;检测波长237nm.结果对乙酰氨基酚血药质量浓度在0.025~25μg·mL-1内的线性关系良好(r=0.9989),血浆中低、中、高3种浓度(0.05,5,20μg·mL-1)的相对回收率在92.9%~92.1%之间;日内RSD≤5.77%,日间RSD≤4.24%.结论本法操作简便、准确、灵敏,适用于乙酰氨基酚的药动学研究.     

HPLC法测人血浆中丹皮酚浓度及其药代动力学研究

目的：建立HPLC丹皮酚胶囊血浓度测定方法，评价丹皮酚胶囊的药代动力学特点。方法：24名健康志愿者单剂量口服160mg丹皮酚胶囊，按设定时间采集肘静脉血经乙睛萃取处理，以XB—C18（250mm×4．6mm，5μm）色谱柱为固定相，四氢呋喃-甲醇-水-磷酸（6：60：34：0．1）为流动相测定丹皮酚血浆浓度。采用DAS2．0计算丹皮酚主要药动学参数。结果：丹皮酚线性范围10—500mg／mL，最低检出浓度为10ng／mL。主要药代动力学参数：Cmax为（116±46）ng／mL，tmax为（1．02±0．13）h，t1/2为1．03h。结论：建立的HPLC方法特异性强、灵敏度高，可用于丹皮酚血药浓度测定和人体药动学研究。     

头孢克洛血药浓度测定及其药代动力学研究

目的建立高效液相色谱法测定人血浆中头孢克洛浓度的方法。方法以头孢拉定为内标,采用DIS-COVERYC18色谱柱(4.6mm×250mm,5μm),以醋酸盐缓冲液-乙腈(83∶17)为流动相,流速:1mL/min;检测波长264nm。结果头孢克洛的线性范围为0.2～30μg/mL,回归方程为Y=0.0913X-0.0094,r=0.9996,最低检测浓度为0.1μg/mL,日内和日间RSD均小于7.5%。结论此法准确简便,适用于头孢克洛药代动力学的研究。     

Method development and validation of almotriptan in human plasma by HPLC tandem mass spectrometry: application to pharmacokinetic study

A simple, sensitive and selective method has been developed for quantification of Almotriptan (AL) in human plasma using Almotriptan-d(6) (ALD6) as an internal standard. Almotriptan and Almotriptan-d(6) were detected with proton adducts at m/z 336.1→201.1 and 342.2→207.2 in multiple reaction monitoring (MRM) positive mode, respectively. The method was linear over a concentration range of 0.5-150.0 ng/mL. The limit of detection (LOD) and limit of quantification (LOQ) for Almotriptan were 0.2 pg/mL and 0.5 ng/mL, respectively. Liquid-liquid extraction was used followed by MS/MS (ion spray). The method was shown to be precise with an average within-run and between-run variation of 0.68 to 2.78% and 0.57 to 0.86%, respectively. The average within-run and between-run accuracy of the method throughout its linear range was 98.94 to 102.64% and 99.43 to 101.44%, respectively. The mean recovery of drug and internal standard from human plasma was 92.12 ± 4.32% and 89.62 ± 6.32%. It can be applied for clinical and pharmacokinetic studies.     

血浆中吡格列酮的测定及其在人体内的药动学研究

目的 建立测定血浆中吡格列酮的方法.方法 采用Diamosil C18(150 mm×4.6 mm,5 μm)色谱柱,以乙腈-pH5.8磷酸盐缓冲液(55:45)为流动相,安定为内标,检测波长225 nm.结果 吡格列酮在0.02-2.00 μg·ml-1范围内,线性良好(r=0.9968),平均回收率为105.3%,日内、日间的RSD为4.23%～18.40%.结论 所建方法操作简便、快速、重复性好、准确可靠,适用于临床血药浓度的监测及药动学研究.     

Validated HPLC method for determination of amlodipine in human plasma and its application to pharmacokinetic studies

A rapid, simple and sensitive high-performance liquid chromatography (HPLC) method has been developed for quantification of amlodipine in plasma. The assay enables the measurement of amlodipine for therapeutic drug monitoring with a minimum detectable limit of 0.2 ng ml(-1). The method involves simple, one-step extraction procedure and analytical recovery was about 97%. The separation was performed on an analytical 125 x 4.6 mm i.d. Nucleosil C8 column. The wavelength was set at 239 nm. The mobile phase was a mixture of 0.01 M sodium dihydrogen phosphate buffer and acetonitrile (63:37, v/v) adjusted to pH 3.5 at a flow rate of 1.5 ml min(-1). The calibration curve was linear over the concentration range 0.5-16 ng ml(-1). The coefficients of variation for inter-day and intra-day assay were found to be less than 10%.     

A new and simple HPLC method for determination of etamsylate in human plasma and its application to pharmacokinetic study in healthy adult male volunteers

A new and simple HPLC assay method was developed and validated for the determination of etamsylate in human plasma. After protein precipitation with 6% perchloric acid, satisfactory separation was achieved on a HyPURITY C₁₈ column (250 mm × 4.6 mm, 5 μm) using a mobile phase comprising 20 mM sodium dihydrogen phosphate-2 hydrate (pH was adjusted to 3.5 by phosphoric acid) and acetonitrile at a ratio of 95:5 v/v. The elution was isocratic at ambient temperature with a flow rate of 0.75 ml/min. Allopurinol was used as internal standard. The calibration curve was linear over the range from 0.25 to 20 μg/ml (r² = 0.999). The limit of quantification for etamsylate in plasma was 0.25 μg/ml. The within day coefficient of variance (%CV) ranged from 3.9% to 10.2%, whereas the between-day %CV ranged from 3.1% to 8.7%. The assay method has been successfully used to estimate the pharmacokinetics of etamsylate after oral administration of a 500 mg tablet under fasting conditions to 24 healthy Egyptian human male volunteers. Various pharmacokinetic parameters including AUC_0–t, AUC_0–∞, C_max, T_max, t_1/2, MRT, Cl/F, and V_d/F were determined from plasma concentration–time profile of etamsylate.     

HPLC法测定人血清美他多辛浓度及药代动力学研究

目的:建立人血清美他多辛浓度的HPLC测定法,进行美他多辛健康志愿者药代动力学研究。方法:志愿者含药血清经去蛋白处理后,以甲醇-5 mmol.L-1乙酸铵(14 86)为流动相,采用Diamon-sil C18柱(250 mm×4.6 mm,5μm)分离,流速1.0 ml.min-1,检测波长286 nm。结果:本法线性范围0.2~12μg.ml-1,最低定量限0.2μg.ml-1。血清中美他多辛绝对回收率73.31%~82.00%,方法回收率91.08%~95.69%,日内RSD≤6.3%,日间RSD≤4.3%。结论:本法简单、快速、灵敏、重现性好,是一种有效的检测人血清美他多辛浓度的方法,适用于人体美他多辛药代动力学研究。     

人血浆中盐酸氨溴索的HPLC测定及其在药动学研究中的应用

［摘要］　目的：建立人血浆中盐酸氨溴索浓度HPFC测定法。方法：采用液-液萃取高效液相色谱紫外检测法，以盐酸尼卡地平为内标，色谱柱为Diamonsil C18柱（150 mm×4.6 mm，5 μm），流动相为甲醇-乙腈-0.01 mol•L-1磷酸盐缓冲液（45∶27∶28），流速为1 mL•min-1，检测波长245 nm。结果：盐酸氨溴索血浆浓度测定方法线性范围为10~480 ng•mL-1，r=0.999 9，定量下限为10 ng•mL-1，方法回收率＞94%。日内和日间RSD＜5%。结论：本试验所建立起来的人血浆中盐酸氨溴索浓度测定法灵敏、准确，可靠，适用于盐酸氨溴索的人体药动学研究。     

HPLC determination of indalpine and its major metabolite in human plasma and a pharmacokinetic application

A high-performance liquid chromatographic method using a fluorometric detector was developed for the determination of plasma concentrations of the antidepressant indalpine (I) and its major metabolite 4-[2-(3-indolyl)ethyl]-2-piperidinone (PK). the same procedure was used to measure, in urine, levels of I and of the metabolite, either conjugated or unconjugated. The sensitivity of the assay is 5 ng ml-1 of plasma or urine for both I and PK. Mean recoveries from plasma for PK, I, and the internal standard (quinine sulfate) were 86.4, 86.8, and 88.5 per cent, respectively. Mean recovery from urine for I was 82.5%. This method was used to establish the pharmacokinetic profiles of I and PK following a single oral administration of I (100 mg) in 8 healthy volunteers. Peak plasma concentrations for I and PK were obtained in an average time of 2.1 and 2.6 h (tmax), respectively. The mean absorption t1/2 of I was 0.8 h, the mean Vda 878 l and the mean clearance 58 l h-1. The mean t1/2 for I and PK was 10.4 and 11.9 h, respectively. In a 12 h urine collection 3 per cent of I was excreted unchanged. No conjugated or unconjugated metabolite was found in urine samples. This method was also used to determine plasma levels 10 h post-dose (50 mg t.i.d.) during chronic oral administration in 20 hospitalized patients. Mean plasma concentrations of I and PK post-dose were 116 ng ml-1 and 43 ng ml-1, respectively.     

固相萃取HPLC测定人血浆中头孢西酮及在药动学研究中的应用

目的:建立反相高效液相色谱办法测定人血浆中头孢西酮浓度,并用于注射用头孢西酮钠人体药代动力学研究。方法:采用高效液相色谱紫外检测法,血浆中加入内标后经固相萃取,色谱柱为 Apollo C_(18)(5μm,250 mm×4.6 mm)。流动相为乙腈-0.02 mol·L~(-1)醋酸铵溶液(18∶82)(pH 5.0),流速1 mL·min~(-1),检测波长为278 nm。结果:本方法线性检测范围为0.5～250μg·mL~(-1),线性关系良好(r=0.9996);最低检测浓度为0.5μg·mL~(-1);方法绝对回收率为67.2%～84.0%,相对回收率为91.1%～102.1%;日内、日间 RSD 均小于8%。结论:本方法灵敏度高,操作简便,可用于人血浆中头孢西酮的浓度测定及临床药代动力学研究。     

Development and validation of UPLC method for determination of primaquine phosphate and its impurities

With the objective of reducing analysis time and maintaining good efficiency, there has been substantial focus on high-speed chromatographic separations. Recently, commercially available ultra-performance liquid chromatography (UPLC) has proven to be one of the most promising developments in the area of fast chromatographic separations. In this work, a new isocratic reverse phase chromatographic method was developed using UPLC for primaquine phosphate bulk drug. The newly developed method is applicable for assay and related substance determination of the active pharmaceutical ingredient. The chromatographic separation of primaquine and impurities was achieved on a Waters Acquity BEH C18, 50 x 2.1mm, 1.7 microm column within a short runtime of 5 min. The method was validated according to the regulatory guidelines with respect to specificity, precision, accuracy, linearity and robustness. Forced degradation studies were also performed for primaquine phosphate bulk drug samples to demonstrate the stability indicating power of the UPLC method. Comparison of system performance with conventional HPLC was made with respect to analysis time, efficiency and sensitivity.     

Determination of nifedipine in human plasma by solid-phase extraction and high-performance liquid chromatography: validation and application to pharmacokinetic studies

Nifedipine, a dihydropyridine calcium channel antagonist, is widely used in the treatment of hypertension and other cardiovascular disorders. A simple, rapid, sensitive, precise and accurate HPLC method, using solid-phase extraction, for the quantitation of nifedipine in human plasma was developed and validated. The calibration graphs were linear in the 5–400 ng/ml concentration range (r>0.999). Recovery for nifedipine was greater than 93.9% and for internal standard nitrendipine was 96.1%. Intra-day and inter-day precision ranged from 1.4 to 4.2 and 3.9 to 5.6%, respectively. Intra-day and inter-day accuracy was ranged from 94.5 to 98.0 and 93.1 to 98.0%, respectively. The method was not interfered with by other plasma components and was applied for the determination of nifedipine in pharmacokinetic study after single oral administration of 10 mg nifedipine to 18 healthy male subjects.     

固相萃取-高效液相色谱法测定人血浆中吡格列酮浓度及其药动学

目的:建立测定人血浆中吡格列酮浓度的高效液相色谱方法。方法:采用Hypersil C_(18)色谱柱,以0.1 mol·L~(-1)醋酸铵-乙腈(305:195)为流动相,罗通定为内标,流速1 mL·min~(-1),检测波长269 nm,血浆样品经固相萃取后直接进样。结果:该法线性范围为50～1600 μg·L~(-1),回归方程为A_s/A_i=0.0127+2.6242×10~(-3) C,r=0.9997(n=5),最低检测浓度为25 μg·L~(-1),提取回收率大于80％,日内日间RSD均小于10％。结论:本法简便灵敏,适用于盐酸吡格列酮血药浓度测定和人体药动学研究。     

高效液相色谱法测定人体地尔硫及其主要代谢物的药物动力学

用高效液相色谱法（ＨＰＬＣ）测定人血清中地尔硫（ＤＺ）及去乙酰地尔硫（Ｍ１）浓度。以ＳｐｈｅｒｉｓｏｒｂＯＤＳ，５μｍ为层析柱，流动相：甲醇∶乙腈∶水（６０∶１０∶３０），检测波长２３７ｎｍ，以盐酸普罗帕酮为内标。检测范围：ＤＺ为５．４５～２７２．５ｎｇ·ｍｌ－１，Ｍ１为５．８５～２９２．５ｎｇ·ｍｌ－１。最低检测浓度：ＤＺ为２．８７ｎｇ·ｍｌ－１，Ｍ１为１．９９ｎｇ·ｍｌ－１。平均回收率ＤＺ为１０１．８８％，Ｍ１为１０１．７２％，ＲＳＤ均在１２％以内。并对４名受试者口服９０ｍｇ盐酸地尔硫片后，其药时曲线经微机用ＰＫＢＰ－Ｎ１程序拟合，ＤＺ为一房室开放模型，Ｍ１为二房室开放模型，求得ＤＺ和Ｍ１的Ｔ１／２分别为５．６±１．５ｈ和１４±７ｈ。     

Development and validation of a HPLC method for the determination of sertraline and three non-chiral related impurities

In this study, a screening on reversed-phase stationary phases (including C₈, C₁₈, CN, PEG and amide) was carried out in order to obtain an efficient HPLC method for the determination of sertraline and three of its more closely related synthetical and non-chiral impurities, without using ion-pair reagents. The best results in terms of both retention time and resolution of the target analytes were obtained with a Zorbax Bonus-RP column, which contains a polar amide group embedded in a C₁₄ alkyl chain.     

RP-HPLC法测定人血浆中拉莫三嗪及药动学

目的建立RP-HPLC法测定人血浆中的拉莫三嗪,并将此方法应用于拉莫三嗪片在健康受试者体内的药动学研究。方法血浆样品经乙酸乙酯提取后,采用Kromasil C18柱分离,流动相为乙腈-20 mmol.L-1 KH2PO4(体积比为25∶75),流速为1.0 mL.min-1,检测波长为306 nm。测定了11名健康受试者口服拉莫三嗪片50 mg后不同时刻血浆中拉莫三嗪的浓度,采用DAS 2.0软件以非房室模型计算药动学参数。结果拉莫三嗪的线性范围为10.0～2 000.0μg.L-1;日内和日间精密度均小于8.0%,准确度(relative error,RE)在±2.6%以内,提取回收率大于59.1%。拉莫三嗪的主要药动学参数:t1/2为(39.1±8.2)h,tmax为(3.3±1.8)h,ρmax为(469.6±152.4)μg.L-1,AUC0-t为(22 424.6±6 952.6)μg.h.L-1,AUC0-∞为(25 573.2±7 196.4)μg.h.L-1。结论该方法适用于拉莫三嗪人体药动学研究。     

HPLC determination of Nimodipine in plasma with an improved sample refining method and its application in pharmacokinetic studies

In order to prepare samples for HPLC analysis with maximum drug recovery and impurity elimination, a revised method for the extraction and purification of a target substance from plasma was developed and applied in a pharmacokinetic study with Nimodipine as a model drug. After protein precipitation of a plasma sample using pure methanol and evaporation of the supernatant to dryness, methanol of various concentrations from 10% to 100% were used to dissolve the remaining residues with the goal of maximizing drug recovery and impurity elimination. Through rigorous screening with HPLC peaks from residual impurity and recovered drug as the criteria, a methanol concentration of 30% was chosen. The standard curve was linear (r²≥ 0.999) over the range of 2–160 ng/mL with a limit of quantification (LOQ) of 2 ng/mL. Intra- and inter-day precision values were below 15%, and the accuracy ranged from –1.70% to 5.88% at all three quality control (QC) levels. The wavelength of maximum absorption was 238 nm, and a smaller LOQ value of 2 ng/mL was achieved compared with the reported method. The revised method was successfully applied in a pharmacokinetic study of Nimodipine in rats and sample preparations of lidocaine hydrochloride.     

RP-HPLC法测定人血浆中厄多司坦浓度及其药代动力学研究

目的:建立厄多司坦血浓度反相高效液相色谱测定方法,进行其人体药代动力学研究。方法:采用单剂两周期交叉试验设计,20名健康男性志愿者随机单剂量口服厄多司坦分散片或胶囊600 mg,按设定时间采集肘静脉血,以 Luna C_(18)(2)(150mm×4.6 mm,5 μm)为固定相,甲醇-醋酸钠(冰醋酸调 pH 至6.8)(5:95)为流动相,流速0.7 mL·min~(-1),检测波长236 nm,测定厄多司坦血浓度,计算其药代动力学参数,评价两制剂的生物等效性。结果:本方法最低定量限为0.01μg·mL~(-1),在0.01～3μg·mL~(-1)(r=0.9991)范围内线性关系良好,高、中、低浓度日内、日间 RSD 均小于5%。厄多司坦分散片和胶囊主要药代动力学参数 t_(1/2)分别为(2.478±1.206)h 和(2.603±1.282)h,T_(max)分别为(1.163±0.424)h 和(1.413±0.424)h,C_(max)分别为(1.419±0.385)μg·mL~(-1)和(1.594±0.558)μg·mL~(-1),AUC_(0～12)分别为(3.416±1.037)μg·mL~(-1)·h 和(3.433±0.910)μg·mL~(-1)·h,AUC_(0～∞)分别为(3.492±1.044)μg·mL~(-1)·h 和(3.523±0.912)μg·mL~(-1)·h。结论:测定方法准确、灵敏、快速,适用于厄多司坦人体药代动力学研究。厄多司坦两制剂主要药代动力学参数无显著性差异,为生物等效制剂。