反相高效液相色谱法同时测定 3 种抗癫痫药物的血药浓 度简

目的建立同时测定咪达唑仑（MID）、硝西泮（NZP）、地西泮（DZP）3种药物血药浓度的反相高效液相色谱法。方法血浆样品以乙酸乙酯＋石油醚提取,色谱柱为SymmetryshieldRP柱（250mm×4．6mm,5．0μm）,流动相为甲醇一磷酸盐缓冲液（55：45,v／v,pH2．15）,流速为0．9mL／min,检测波长为230nm,柱温为26℃,进样量为20μL。结果MID,NZP,DZP质量浓度均在20—4000ng／mL范围内与峰面积呈良好线性关系（r≥0．9996,n=5）;最低检测浓度（S／N=3）,NZP,DZP为10ng／mL,MID为20ng／mL;回收率均在98．2％～103．9％之间,日内及日间RSD均〈5％。结论该法能够同时测定3种药物的血药浓度,且简便、准确,适用于临床血药浓度监测。     

人血浆中特非那定的LC—MS测定法

目的：建立人血浆中特非那定的高效液相色谱．电喷雾质谱联用（HPLC／ESI—MS）测定方法。方法：以氟桂利嗪为内标,血浆以2mol／LNaOH溶液碱化后,经乙酸乙酯提取;选用PhenomenexC18色谱柱（250mm×4．6mm,5／zm）,柱温25℃;流动相为甲醇．水80：20（含1％的冰醋酸和0．5％的三乙胺）,流速1mL／min;采用四级杆电喷雾质谱检测器进行定量,选择性正离子检测（SIM）,特非那定m／z472,氟桂利嗪m／z203。结果：本法线性范围为0．1～20ng／mL,最低检测限为0．05ng／mL（S／N〉3）,血浆中特非那定的日内和日间精密度为2．54％～9．28％,回收率为88．62％～91．67％。结论：本法专属性强、灵敏度高,适用于临床试验中特非那定的药代动力学研究。     

HPLC—MS／MS法测定人体血浆中格列美脲及其活性代谢物

目的：建立同时测定格列美脲（Glimepiride,G）及其活性代谢物羟基格列美脲（Hydroxyl—glimepiride,M1）血浆浓度的高效液相色谱一串联质谱（HPLC—MS／MS）法。方法：血浆样品酸化后,经乙酸乙酯萃取后浓集进样,色谱柱为PhenomenexGeminiC18（50mill×3．0mm,5μm）,甲醇：水：甲酸（80：20：0．1）为流动相,流速为0．2mL／min,柱温为室温,采用电喷雾（ESI）离子源,多反应离子检测模式,以格列齐特（Gliclazide,IS）为内标。G、M1和IS的检测离子对分别为质荷比（仇／z）491．4—352．4、507．4—352．4、324．4—127．2。结果：G和M1的线性范围分别为1．25～400ng／mL和0．313～100ng／mL,最低定量限分别为1．25和0．313ng／mL;批内批间精密度均小于10％,方法回收率均在96．4％～102．5％之间。结论：该方法简便快速、特异性高,可用于同时测定G和M．血药浓度及其人体药代动力学的研究。     

RP-HPLC法测定法罗培南血药浓度和尿药浓度

目的：建立测定法罗培南血药浓度和尿药浓度的RP-HPLC法。方法：采用Diamond C18色谱柱（250mm×4．6mm,5μm）,血样与尿样测定流动相分别为0．02mol／L磷酸二氢钾缓冲液（pH=2．95）：乙腈=75：25和81：19,流速均为1．0mL／min,检测波长均为318nm。结果：血浆样品的线性范围为0．03125～2．5μg／mL（r=0．9999）、2．5～120μg／mL（r=0．9997）,最低检测浓度为31．25ng／mL,绝对回收率为68．9％～77．8％;尿液样品的线性范围为0．5～20μg／mL（r=0．9999）、20～1600μg／mL（r=0．9997）,最低检测浓度为0．5μg／mL,绝对回收率为96．5％～106．0％。日内及日间精密度均〈11．0％。结论：本方法简单、快速,灵敏度和准确度较高,能满足法罗培南钠注射液临床药动学研究的需要。     

大鼠血浆五味子甲素、乙素、醇甲和酯甲高效液相色谱串联质谱测定方法的建立

目的：建立大鼠血浆中五味子甲素、乙素、醇甲和酯甲的高效液相色谱串联质谱（LC-MS／MS）分析方法，为五味子提取物中四种主要成分的药代动力学研究提供理想的测定手段。方法：采用高效液相色谱串联线性离子阱质谱同时检测五味子提取物中四种主要成分，采用电喷雾（ESI）离子源，多反应监测模式（MRM）进行检测。色谱柱为ZORBAX SB-C18（2．1×100mm，3．5μm），流动相为甲醇-水（75：25，0．1％甲酸），流速为0．2mL／min。双环醇（BY）为内标（IS），血浆样品经甲醇沉淀、高速离心后进样分析，各成分监测离子分别为：甲素m／z 439→393，乙素m／z 423→377，醇甲m／z 455→409，酯甲m／z 559→437，BYm／z 413→399。结果：五味子甲素、醇甲、乙素、酯甲在0．01～2．0μg／mL的浓度范围内线性关系良好，，最低定量限为0．01～0．02／mL，R2〉0．99。五味子四种成分的日内和日间精密度RSD均〈15％，绝对回收率〉80％。结论：本研究成功建立了大鼠血浆中五味子甲素、乙素、醇甲和酯甲的高效液相色谱串联质谱定量方法，该方法专属性强、快速、灵敏、可靠，可满足五味子提取物中四种主要成分药代动力学研究的要求。     

LC-MS／MS法测定人血浆中的帕珠沙星

目的：建立一种快速灵敏检测帕珠沙星血药物浓度的液相色谱－串联质谱方法。方法色谱柱为 Agilent Eclipse XDB－C18（4．6mm ×150mm,5μm）;流动相为乙腈：水（0．005mol· L－1甲酸铵,0．1％甲酸）＝（30∶70）;流速：0．5mL· min －1;质谱条件为电喷雾电离源（ESI）;采用选择反应监测（SRM）对帕珠沙星（m／z 319．0→301．1）和环丙沙星（m／z 332．2→314．2）进行测定。样本处理采用乙腈沉淀蛋白法,吸取上清液1μL进样。结果帕珠沙星的线性范围为0．02～20μg· mL －1,提取回收率为82％～92％,日内和日间精密度均小于15％。结论本方法简便、灵敏,该方法经考察符合血浆样品的测定要求,可以应用于临床药物浓度的测定和药代动力学的研究。     

高效液相色谱串联质谱联用法测定人血浆中格拉司琼的浓度

目的建立液相色谱-电喷雾串联质谱法测定人血浆中格拉司琼的浓度。方法血浆样品经乙腈直接沉淀蛋白后，采用Thermo Hypurity C18柱（2．1mm×150mm，5μm）分离，流速为0．2mL·min^-1，以乙腈-0．1％甲酸=50：50为流动相等度洗脱。通过电喷雾离子源进入质谱，以二级质谱多反应监测（MRM）方式进行检测。结果格拉司琼的线性范围为0．11～109ng·mL^-1，最低检测浓度为0．11ng·mL^-1，平均相对回收率在91．30％～105．46％，日内和日间变异均〈15％。结论本法简单、快速、灵敏、重现性好，可用于格拉司琼的人体药物动力举和硅物隼静桂研雾．     

UPLC／Q—TOFMS法测定大鼠灌胃人工麝香后血浆中麝香酮的浓度

目的：建立检测麝香酮在灌胃人工麝香大鼠体内的血药浓度的分析方法,用于人工麝香的药动学研究。方法：血浆样品以乙醚液-液萃取法进行前处理,采用超高效液相色谱与串联四级杆飞行时间质谱仪联用技术（UPLC／Q—TOFMS）进行分析。ACQUITY UPLC BEH C18色谱柱（2．1mm×100mm,1．7μm）,柱温35℃,流动相为乙腈：水（均含0．1％的甲酸85：15）,流速0．2mL／min。使用APCI离子源,正离子模式下采集数据。结果：麝香酮血浆浓度在0．0756～3．78μg／mL范围内线性关系良好（R=0．9980）,最低检测限为0．05μg／mL。高、中、低三种浓度的准确度和精密度均符合生物样品检测要求（RE〈10％,RSD〈15％）,平均绝对回收率均〉81％。结论：本实验所建立的方法稳定可靠,专属性强,灵敏度较高,测定方便快速,适用于人工麝香中麝香酮的临床前血药浓度分析。     

用RP—HPLC法测定人体内辛伐他汀血药浓度

目的：建立RP-HPLC法测定人血浆中辛伐他汀的含量。方法：采用Kromasil C18色谱柱（250mm×4．6mm，5μm）；流动相为甲醇-乙腈-水（80：5：15）；流速为1．0mL／min；检测波长238nm；柱温为30℃。结果：辛伐他汀的血药浓度在2．0～150．0ng／mL范围内线性良好，定量下限为2．0ng／mL。回归方程为A=1．589×10^8c-218．0，r=0．9999（n=7），低、中、高浓度（6．5、23．0、65．0ng／mL）的相对回收率分别为（100．28±3．35）％、（97．57±3．21）％和（100．25±2．82）％，日内和日间RSD为2．40％～4．32％（n=5）。结论：本法快速、高效、灵敏，适用于辛伐他汀的血药浓度测定。     

超高效液相色谱串联质谱法测定人血浆中阿托伐他汀浓度

目的建立测定人血浆中阿托伐他汀浓度的超高效液相色谱串联质谱（UPLC—MS／MS）法。方法色谱柱为Acquity UPLC BEH C18柱（50mm×2．1mm。1．7μm）,柱温40℃,流动相为乙腈-含0．01％甲酸的水溶液（70：30）,流速为0．3mL／min,进样量10μL,电喷雾离子化（ESI^＋）正离子MRM扫描。检测离子为m／z 559．2[M＋H]^＋→440．5[M＋H]^＋（阿托伐他汀）,m／z315．5[M＋H]^＋→109．3[M＋H]^＋（黄体酮）,血浆样品采用乙酸乙酯提取,50℃水浴氮气挥干,用黄体酮作为内标定量分析。结果阿托伐他汀血药浓度在0．01。50ng／mL线性关系良好（r=0．9989）。回收率在95％-106％之间,萃取回收率在72％-79％之间（RSD〈4％,n=5）,日内、日间精密度的RSD均小于5％（n=5）,最低检测质量浓度为0．002ng／mL。结论该方法准确、灵敏、简便,适用于阿托伐他汀的血药浓度测定。     

A simple and sensitive liquid chromatography–mass spectrometry routine method for determination of clarithromycin in human plasma: Application to a clinical pharmacokinetic study

A simple and sensitive liquid chromatography-mass spectrometry (LC-MS) method was developed for the routine determination of Clarithromycin in human plasma, with Alprazolam as the internal standard. The separation was carried out on an Phenomenex (C18 100?×?4.6?mm, i.d 5 µm) column, with Acetonitrile and 10?mM Ammonium acetate Buffer solution (95:5) as the mobile phase under isocratic conditions at a flow rate of 0.2?mL/min. Detection of the analytes was achieved using positive ion electrospray tandem mass spectrometry in selected reaction monitoring (SRM) mode. The standard curves were linear (r²?>?0.99) over the concentration range of 300–5000?ng/mL. The intra- and inter-day precision (RSD) values were below 15%. The method is validated for the estimation of Clarithromycin in human plasma over a concentration range of 300–5000?ng/mL. Expected recoveries were observed. Limit of detection of the methods is 3.0?ng/?mL and limit of quantitation is 10.0?ng/mL that shows that the developed method has adequate sensitivity. The method was successfully applied to the pharmacokinetic study of Clarithromycin tablets in healthy male volunteers after oral administration.     

Rapid determination of tramadol in human plasma by headspace solid-phase microextraction and capillary gas chromatography-mass spectrometry

A simple, rapid and sensitive method for determination of tramadol in plasma samples was developed using headspace solid-phase microextraction (HS-SPME) and gas chromatography with mass spectrometry (GC-MS). The optimum conditions for the SPME procedure were: headspace extraction on a 65-microm polydimethylsiloxane/divinylbenzene (PDMS/DVB) fiber; 0.5 mL of plasma modified with 0.5 mL of sodium hydroxide (0.1 M); extraction temperature of 100 degrees C, with stirring at 2000 rpm for 30 min. The calibration curve showed linearity in the range of 1-400 ng mL(-1) with regression coefficient corresponding to 0.9986 and coefficient of the variation of the points of the calibration curve lower than 10%. The detection limit for tramadol in plasma was 0.2 ng mL(-1). The proposed method was successfully applied to determination of tramadol in human plasma samples from 10 healthy volunteers after a single oral administration.     

Gradient elution liquid chromatography mass spectrometry determination of acetylcorynoline in rat plasma and its application to a pharmacokinetic study

Abstract

1.?Acetylcorynoline is the major alkaloid component derived from Corydalis bungeana herbs. A sensitive and selective liquid chromatography mass spectrometry method for determination of acetylcorynoline in rat plasma was developed over the range of 5–1000?ng/mL to characterize the pharmacokinetic properties.

2.?Chromatographic separation was achieved on a C18 (2.1?mm×?150?mm, 5?μm) column with acetonitrile 0.1% formic acid in water as mobile phase with gradient elution. The flow rate was set at 0.4?mL/min. After addition of carbamazepine as internal standard (IS), protein precipitation by acetonitrile–methanol (9:1, v/v) was used as sample preparation. An electrospray ionization source was applied and operated in positive ion mode; selective ion monitoring mode was used for quantification using target ions m/z 410 for acetylcorynoline and m/z 237 for the IS.

3.?Mean recoveries of acetylcorynoline in rat plasma were in the range of 72.3–87.6%. Matrix effects for acetylcorynoline were measured to be between 88.7% and 93.5%. Coefficient of variation of intra-day and inter-day precision were both <13%. The accuracy of the method ranged from 95.8% to 112.1%. The analyte was stable under auto-sampler, room temperature, freeze-thaw and long-term (20 days), the bias in concentration was within ±15% of their nominal values.

4.?The LC-MS method for the determination of acetylcorynoline in rat plasma utilizing 100?µL of plasma with an LLOQ of 5.0?ng/mL developed and validated, it was sensitive, selective and simple. This method was successfully applied in pharmacokinetic study of acetylcorynoline after intravenous administration of single dosage 3?mg/kg in rats.     

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。     

高效液相色谱法测定人血浆中酮咯酸浓度

目的建立测定人血浆中酮咯酸浓度的高效液相色谱法。方法 24例健康志愿者口服单剂量(20 mg)酮咯酸氨丁三醇胶囊试验制剂。以奥硝唑为内标,待测血浆用甲醇沉淀法进行处理,色谱柱采用PlatisilODS柱(250 mm×4.6 mm,5μm),流动相为0.05 mol/L磷酸二氢钠溶液-乙腈(70∶30)(2 mol/L氢氧化钠调节pH值至5.14±0.01),波长为330 nm,进行HPLC分析。结果试验制剂的主要药动学参数:t1/2为(6.08±1.82)h,Cmax为(1 351.7±310.9)ng/mL,Tmax为(1.86±0.58)h,AUC0-t为(7 735±1 718)ng.h/mL,AUC0-∞为(8183±1 939)ng.h/mL,无内源性物质干扰测定,酮咯酸在24.8～2 480.0 ng/mL范围内与内标物峰面积之比线性关系良好,定量下限为24.8 ng/mL。提取回收率为83.9%～94.6%。结论该方法操作简便、准确,灵敏度高,专属性强,适用于人血浆中酮咯酸浓度的测定。     

Determination of hydrochlorothiazide in human plasma by liquid chromatography/tandem mass spectrometry

In this study, a fast and sensitive liquid chromatography/tandem mass spectrometry method for determination of hydrochlorothiazide in human plasma was developed and validated. The analyte and irbesartan, used as the internal standard, were precipitated and extracted from plasma using methanol. Analysis was performed on a Phenomenex Kromasil C₈ column with water and methanol (27:73, v/v) as the mobile phase. Linearity was assessed from 0.78 to 200 ng/mL in plasma. The analytical method proved to be applicable in a pharmacokinetic study after oral administration of 12 mg hydrochlorothiazide tablets to 20 healthy volunteers.     

Hydrophilic interaction chromatography-tandem mass spectrometry of donepezil in human plasma: Application to a pharmacokinetic study of donepezil in volunteers

A selective, sensitive and rapid hydrophilic interaction liquid chromatography with electrospray ionization tandem mass spectrometry was developed for the determination of donepezil in human plasma. Donepezil was twice extracted from human plasma using methyl tert-butyl ether at basic pH. The analytes were separated on an Atlantis HILIC Silica column with the mobile phase of acetonitrile: ammonium formate (50 mM, pH 4.0) (85:15, v/v) and detected by tandem mass spectrometry in the selective reaction monitoring mode. The calibration curve was linear (r = 0.9994) over the concentration range of 0.10-50.0 ng/mL and the lower limit of quantification was 0.1 ng/mL using 200 muL plasma sample. The coefficient of variation and relative error for intra-and inter-assay at four QC levels were 2.7 to 10.5% and -10.0 to 0.0%, respectively. There was no matrix effect for donepezil and cisapride. The present method was successfully applied to the pharmacokinetic study of donepezil after oral dose of donepezil hydrochloride (10 mg tablet) to male healthy volunteers.     

Liquid chromatography-tandem mass spectrometry method for the determination of trimetazidine in human plasma

A sensitive liquid chromatography-tandem mass spectrometry (LC/MS/MS) method was developed and validated for the determination of trimetazidine (CAS 13171-25-0) in human plasma, using pseudoephedrine as internal standard (IS). Plasma samples were simply pretreated with methanol for deproteinization. The chromatographic separation was performed on a C18 column with a mobile phase of 3 mmol/L ammonium acetate solution-methanol (15:85, v/v) at a flow rate of 0.3 mL/min. The chromatographic separation was achieved in less than 3.2 min. The linearity was established over the concentration range of 1-100 ng/mL. Both intra- and inter-batch standard deviations were less than 9.5%. The method was successfully applied to study the relative bioavailability of trimetazidine hydrochloride tablets in healthy Chinese volunteers and the pharmacokinetic parameters of the reference and test tablets were compared.     

Quantitative determination of doxorubicin in the exosomes of A549/MCF-7 cancer cells and human plasma using ultra performance liquid chromatography-tandem mass spectrometry

In cancer therapy, exosomes efflux enhances resistance of cancer cells toward anticancer agents through mediating the transport of anticancer drugs outside the cells. In this study, a rapid, simple and highly sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the determination of Doxorubicin (DOX) in exosomes of cancer cells and human plasma using Ketotifen as an internal standard (IS). Plasma samples spiked with DOX and two cancer cell lines (A549 & MCF-7) were incubated with different concentrations of DOX and IS. The analytes were then extracted with methanol after protein precipitation and the chromatographic separation was carried out using a C18 column, with a mixture of acetonitrile–water- formic acid (85:15:0.1%, v/v/v) as mobile phase. Multiple reaction monitoring (MRM) was utilized to monitor the protonated precursor to product ion transitions of m/z 544.25?>?397.16 and m/z 310.08?>?96.97 for the quantification of DOX and IS, respectively. The method was linear over ranges of 1–1000?ng/mL for DOX in plasma and 2–1000?ng/mL for DOX in exosome samples. The lower limit of quantification of this method was 1?ng/mL, 2?ng/mL and 2?ng/mL in human plasma, A549 & MCF-7 cells respectively. Intra- and inter day precision of all quality control concentrations were less than 10.33% and the accuracy values ranged from ?4.82 to 12.60%. The optimized UPLC-MS/MS method proved to be fast, specific, simple and highly sensitive and was successfully applied for the estimation of DOX in the exosomes of cancer cells and plasma.     

高效液相色谱梯度洗脱法测定克拉霉素的含量

目的建立高效液相色谱梯度洗脱法测定克拉霉素的含量。方法采用kromasil-C18(4.6mm×150mm,5μm)柱,乙腈-0.476%磷酸二氢钾溶液(pH值4.4)为流动相梯度洗脱,检测波长为205nm,流速1.1mL/min。结果克拉霉素在0.49～2.48mg/mL内呈良好的线性相关性,相关系数r为0.9999,检测限为0.8ng/mL,定量限为2.4ng/mL。结论该方法准确可靠,灵敏度高,可用于克拉霉素的含量测定及质量控制。