液相色谱-串联质谱法测定人血浆中的利培酮

利培酮为苯并异噁唑类化合物,通过阻断5-HT2受体和多巴胺D2受体发挥抗精神病作用.同其他抗精神病药物相比,利培酮所引起的椎体外系副作用更少,药效更为明显 .     

亲水作用色谱-串联质谱法测定气雾剂给药后人血浆中的沙丁胺醇北大核心CSCD

目的:建立灵敏、快速的亲水作用色谱-串联质谱法(HILIC-MS/MS)测定给予沙丁胺醇气雾剂后人血浆中的沙丁胺醇。方法:以d9-沙丁胺醇为内标,血浆样品经乙腈蛋白沉淀后,采用Luna HILIC(100 mm×3.0 mm,3μm)色谱柱分离。流动相为乙腈(含0.1%甲酸)-5 mmol.L-1醋酸铵溶液(氨水调节pH为8.5)(93∶7,v/v),等度洗脱,进样体积2.0μL,样品分析时间为3.5 min。采用电喷雾电离(ESI)源正离子模式、多反应监测(MRM),用于定量分析的离子反应分别为m/z 240→(148+166)(沙丁胺醇)和m/z 249→(149+167)(d9-沙丁胺醇)。结果:沙丁胺醇定量方法的线性范围为11.7～2340 pg.mL-1,定量下限达11.7 pg.mL-1,日内、日间精密度(RSD)均小于14.1%,准确度(RE)在-3.8%～-0.8%之间。本法成功应用于健康受试者给予硫酸沙丁胺醇气雾剂200μg后的药动学研究。结论:采用稳定同位素内标的HILIC-MS/MS法快速、灵敏和准确,适用于测定人血浆样品中的沙丁胺醇,可测定给药后24 h的血浆浓度。     

液相色谱-串联质谱法测定人毛发中的美沙酮

目的:建立人毛发中美沙酮的液相色谱-串联质谱检测方法。方法:人毛发经清洗研磨后,在pH 9.2的硼酸缓冲液中超声90 min,固相萃取法提取,用LC-MS/MS检测。结果:人毛发中美沙酮的检出限为0.01 ng.mg-1,线性范围为0.1-100 ng.mg-1(r=0.9988),回收率96.7%-118.6%,日内及日间精密度<10%。用此法测定人毛发阳性对照品,结果满意。结论:本方法灵敏度高,适用于人毛发中美沙酮的测定。     

高效液相色谱-串联质谱法测定血清中的雌激素

目的 建立同位素内标-高效液相色谱-串联质谱方法测定人血清中3种雌激素的含量。方法 采用高效液相色谱-串联质谱仪,色谱柱为Phenomenex Gemini^?3μm C₁₈ 110?(50 mm×3 mm),柱温40℃。流动相为0.1%氨水溶液:甲醇,流速为0.6 ml/min。在电喷雾负离子模式下,采用多反应监测模式对血清中雌激素含量进行定量。结果 3种雌激素在相应的浓度范围内相关系数r均大于0.997,加标回收率为91.76%～105.90%,精密度为0.25%～3.04%,无携带污染。结论 该方法准确、可靠、灵敏度好、专属性强,适用于血清中雌激素的检测。     

液相色谱串联质谱法测定毛发中的甲睾酮浓度

目的：建立测定毛发中甲睾酮浓度的液相色谱-串联质谱（LC—MS／MS）法。方法：头发样品经氢氧化钠溶液消解、戊烷液-液提取后，采用Agilent ZORBAXSB—C18柱（150mm&#215;2．1mm，5μm）分离，流动相为甲醇-20mmol&#183;L^-1乙酸铵和0．1％甲酸溶液（73：27），流速为0．2mL&#183;min^-1样品在三重四极杆串联质谱中经电喷雾正离子源离子化后，以多反应离子监测方式（MRM）测定。结果：甲睾酮浓度在10～1000pg&#183;mg^-1范围内线性良好（r=0．9999），检测限为5pg&#183;mg^-1,回收率为80．6％～112．9％，日内、日间变异（RSD）均≤15％。结论：方法简便、快速、灵敏、准确，专属性强，适用于毛发中甲睾酮浓度的测定。     

用液相色谱-串联质谱法测定血浆总同型半胱氨酸

目的：建立血浆中总同型半胱氨酸(tHcy)的液相色谱-串联质谱(LC-MS／MS)测定法。方法：血浆tHcy及内标氘代高胱氨酸经巯基乙醇还原后，用10％三氯乙酸沉淀血浆蛋白，经液相色谱分离后，采用电喷雾离子化-串联质谱进行多反应监测(MRM)检测，离子选择通道分别为m／z：136．2／46．9(Hcy)，140．2／48．9(Hcy-d4，氘化Hcy)。Hcy、Hcy-d4的保留时间均为6.3min。结果：Hcy在2．5～640μmol／L范围内线性关系良好。本法的最低检测浓度为O．3μmol／L(S／N≥5)。批内、批间精密度分别为1.97％～8.79％、2．55％～4．14％。方法回收率为100．3％～109．1％。结论：该方法的预处理快速、简便，检测专一灵敏，试剂成本低廉，可适应开展大规模临床Hcy研究的需要。     

高效液相色谱-串联质谱法测定人血浆中的缬沙坦

目的:建立灵敏、快速的高效液相色谱-串联质谱测定人血浆中缬沙坦浓度的方法,并用于人体药代动力学研究。方法:200μL的血浆样品经乙醚提取处理后,以乙腈-水-甲酸(70∶30∶0.2)为流动相,Diamonsil-C18柱分离,通过电喷雾离子源四极杆串联质谱,以多反应监测(MRM)的方式进行检测,选择监测离子反应为m/z436.3→291.2(缬沙坦)和423.1→207.1(氯沙坦)。结果:缬沙坦在2.120~5 300 ng.mL-1浓度范围内线性关系良好,定量限为2.120 ng.mL-1,日内、日间精密度(RSD)均≤9.0%,准确度在0.9%以内,缬沙坦和内标氯沙坦的回收率分别为83.1%和79.4%。结论:该法选择性强、灵敏度高,适用于缬沙坦的临床药动力学研究。     

液相色谱-串联质谱法测定柏子仁中黄曲霉毒素残留量

目的建立检测柏子仁中黄曲霉毒素B1,B2,G1,G2残留量的液相色谱-串联三重四极杆质谱法。方法将样品提取液经免疫亲和柱净化,甲醇洗脱,以甲醇/乙腈-0.1 mmol乙酸铵为流动相,经反相色谱柱Agilent Zorbax Extend-C18柱(250 mm×4.6 mm,5μm),用电喷雾正离子二级离子监测扫描模式(MRM)检测。结果黄曲霉毒素G2和B2质量浓度在0.03～3.2 ng/mL范围内、黄曲霉毒素B1和G1在0.1～10 ng/mL范围内与峰面积积分值线性关系良好。检测限(LOD)为0.1 ng/mL,定量限(LOQ)为0.2 ng/mL。3个质量浓度(0.25,1.0,5.0 ng/mL)的加标平均回收率为84.1%～96.8%,RSD为7.7%～13.4%。结论所用方法专属性强、灵敏度高、简便快速、结果准确,可定性定量测定柏子仁中残留的黄曲霉毒素。     

Determination of metformin and its prodrugs in human and rat blood by hydrophilic interaction liquid chromatography

Simple and specific hydrophilic interaction liquid chromatography (HILIC) method with ultraviolet (UV) detection was developed for the simultaneous determination of highly water-soluble metformin and its more lipophilic prodrugs in human and rat blood samples. The sample preparation was accomplished by precipitating proteins with acetonitrile, which enabled the direct injection of supernatants to the HPLC. Chromatographic separation was performed on an analytical normal phase silica column using a mixture of 0.01 M ammonium acetate pH 5.0 and acetonitrile (40:60, v/v) as a mobile phase at flow rate of 1 ml/min and at the wavelength of 235 nm. The method was validated in terms of specificity, linearity, accuracy, precision, recovery, and analyte stability. The UV-HILIC method was suitable for detecting both metformin and one of its more lipophilic prodrugs simultaneously in human and rat blood samples.     

Analysis of cephalosporins by hydrophilic interaction chromatography

A simple hydrophilic interaction chromatography (HILIC) method was developed to analyze seven cephalosporins. These seven cephalosporins could be separated well on the Click β-CD column and Atlantis HILIC Silica column. The effects of buffer concentration and pH on the retention under HILIC mode were studied. Except cefepime hydrochloride (4), the retention of other six cephalosporins increased with increasing buffer concentration, while decreased with increasing pH. Different separation selectivities could be observed on the Click β-CD column and Atlantis HILIC Silica column, and changing pH also resulted in the changing of separation selectivity. The separations of cephalosporins by HILIC and reversed-phase high performance liquid chromatography (RP-HPLC) were compared, and the two separation modes had good orthogonality. In addition, cefotaxime sodium (1) and its degradation were separated well on the Click β-CD column, which indicated that the Click β-CD column by HILIC can be used for studying the stability of cephalosporins.     

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.     

液相色谱串联质谱法测定人血浆中替考拉宁的血药浓度

目的:建立测定人血浆中替考拉宁浓度的液相色谱串联质谱(LC-MS/MS)测定方法。方法:选用Welch Ultimate XB-C₁₈(2.1 mm×50 mm,5.0 μm)色谱柱,以达托霉素为内标,以含0.1%甲酸的水溶液和乙腈为流动相进行梯度洗脱,柱温为40 ℃。利用乙腈进行沉淀蛋白样品前处理,采用电喷雾离子化多反应监测正离子模式,测定替考拉宁A_2-1、A_2-2、A_2-3、A_2-4和A_2-5 5种主要成分的总浓度。结果:替考拉宁的总浓度线性范围为1~60 μg·mL^-1,定量下限为1 μg·mL^-1,日内和日间精密度RSD均小于15%,提取回收率80.04%~96.02%,内标归一化基质效应为90.12%~104.04%。研究测定了16例给予替考拉宁后患者的血药谷浓度,浓度范围从定量下限至31.2 μg·mL^-1,达标率56.25%,患者个体差异大且初始达标率低。结论:该研究建立了一种快速、简便、稳定的LC-MS/MS测定人血浆中替考拉宁血药浓度的方法,具有较高准确度、精密度和回收率,为临床给药方案的优化奠定基础。     

亲水作用色谱串联质谱法测定人血浆中的拉米夫定

目的:建立简便、快速的亲水作用色谱串联质谱法(HILIC-MS/MS)测定人血浆中的拉米夫定。方法:以13C1,15N2(拉米夫定为内标,血浆样品经乙腈沉淀蛋白后,采用Luna HILIC(100 mm×3.0 mm,3μm)柱分离。流动相为乙腈-5 mmol·L-1醋酸铵-甲酸(95∶5∶0.01,v/v/v),进样体积2μL,样品分析时间3 min。采用ESI源正离子模式、多反应监测(MRM),用于定量分析的离子反应分别为m/z230(112(拉米夫定)和m/z233(115(内标)。结果:测定人血浆中拉米夫定的线性范围为8.0～2000 ng·mL-1,定量下限为8.0 ng·mL-1,日内、日间精密度(RSD)均小于9.0%,准确度(RE)在-7.1%~2.7%之间。本法成功应用于健康受试者口服2种拉米夫定片后的生物等效性研究。结论:采用稳定同位素内标的HILIC-MS/MS法更为简便、快捷和准确,适用于人血浆样品中拉米夫定的测定。     

液相色谱串联质谱法分析血浆中美金刚浓度及其应用

目的：建立一种新的液相色谱质谱法（LC—MS／MS）,测定人血浆中关金刚的浓度。方法：血浆样品经萃取后,以C18柱进行反相分离;采用电喷雾电离源,以多反应监测（MRM）方式进行正离子检测。结果：血浆中关金刚测定线性范围为0．2-30μg·L^-1,最低定量限（LIDQ）为0．2μg·L^-1,r均大于0．99。日内和日间精密度均小于15％,绝对回收率大于79％。结论：该改良法选择性强、灵敏度高、重现性好,能快速、准确测定人血浆中关金刚浓度并成功应用于评估美金刚生物等效性的研究。     

Quantification of three beta‐lactam antibiotics in breast milk and human plasma by hydrophilic interaction liquid chromatography/positive‐ion electrospray ionization mass spectrometry

The use of cephalosporins during breast feeding raises several issues, including the risk of drug exposure through breast milk for the infant. In this paper, a hydrophilic interaction liquid chromatography/positive ion electrospray mass spectrometric assay (HILIC/ESI‐MS) was developed for the quantitation of cefuroxime, cefoxitin, and cefazolin in breast milk and human plasma. The assay was based on the use of small sample size, 25 μL of biological samples, following acetonitrile precipitation of proteins and filtration that enabled injection into the HILIC/ESI‐MS system. All analytes and the internal standard, alfuzosin, were separated by using a ZIC®‐HILIC analytical column (150.0 × 2.1 mm i.d., particle size 3.5 µm, 200 Å) with isocratic elution. The mobile phase was composed of a 6% 12.5 mM ammonium acetate water solution in acetonitrile and pumped at a flow rate of 0.25 mL min^‐1. The assay was linear over a concentration range of 0.2 to 5 µg mL^‐1 and 0.4 to 20 µg mL^‐1 for all the analytes in breast milk and in human plasma, respectively. Intermediate precision was found to be less than 4.2% over the tested concentration ranges. A run time of less than 12 min for each sample made it possible to analyze a large number of biological samples per day. The method is the first reported application of HILIC in the analysis of antibiotics in breast milk and human plasma and it can be used to support a wide range of clinical studies. Copyright © 2016 John Wiley & Sons, Ltd.     

LC-MS/MS法测定人血浆中替加色罗的浓度及其

目的:建立灵敏、快速的LC-MS/MS法测定人血浆中替加色罗,并用于制剂生物等效性研究。方法:血浆样品经有机溶剂提取后,以乙腈-5 mmol.L-1乙酸铵-甲酸(70∶30∶0.01)为流动相,采用Zorbax XDB-C18柱(150 mm×4.6 mm,5μm)分离,通过电喷雾离子化四极杆串联质谱,以多反应监测(MRM)方式进行检测。用于定量分析的离子反应分别为m/z302→173(替加色罗)和m/z256→167(内标苯海拉明)。结果:替加色罗测定方法的线性范围为0.010~10 ng.mL-1,定量下限为0.010 ng.mL-1,日内、日间精密度(RSD)均<7.3%,准确度(RE)在±1.4%之内。应用此法研究比较了22例健康受试者单剂量口服替加色罗参比制剂和受试制剂6 mg后的主要药动学参数,受试制剂和参比制剂Tmax分别为(0.86±0.22)和(1.01±0.24)h,Cmax分别为(2.21±0.69)和(2.05±0.64)ng.mL-1,t1/2α分别为(1.18±0.44)和(1.24±0.56)h,t1/2β分别为(10.10±3.07)和(8.81±2.35)h,AUC0~36 h分别为(6.35±2.48)和(6.47±1.99)ng.h.mL-1,AUC0~∞分别为(6.69±2.59)和(6.70±2.03)ng.h.mL-1。马来酸替加色罗分散片的相对生物利用度平均为(98.2±22.1)%。结论:该法灵敏、快速、准确,适用于替加色罗制剂的人体生物等效性评价。     

固相萃取亲水作用色谱法测定人体血浆中表阿霉素

目的:建立固相萃取-亲水作用色谱法(SPE—HILIC)测定人体血浆中的表阿霉素。方法:血浆样品中加入表柔红霉素作为内标,经 Oasis HLB 固相萃取(SPE)小柱萃取后进样测定,色谱柱为 Kromasil KR100—5SIL 硅胶色谱柱(250 mm×4.6mm,5μm),乙腈-甲酸铵缓冲液(40 mmol·mL~(-1),pH 2.9)(90:10)为流动相,检测波长为254 nm。结果:血浆中表阿霉素的线性范围为0.05～2.5μg·mL~(-1)(r~2=0.9991);最低检测限为0.05μg·mL~(-1);样品的回收率高于89.4%;日内及日间精密度RSD 小于7.0%。结论:方法简便,准确可靠,流动相与质谱检测器兼容,适用于血浆表阿霉素浓度测定及药代动力学研究。     

亲水作用色谱法（HILIC）的方法开发策略及其在铂类抗癌药分析中的应用

亲水作用色谱法解决了大多数极性化合物的色谱分离问题,其已成为了包括铂类抗癌药在内的许多极性化合物的色谱分离的首选。亲水作用色谱法在极性化合物的分离与检测、药代动力学研究等的应用均日益广泛。如何高效的完成亲水作用色谱的方法开发是药物分析科学家和药代动力学科学家都面临的重要问题,然而目前还没有专门的文献对此进行系统化的整理和研究。本文以此为切入点,围绕亲水作用色谱法方法开发中的固定相选择、流动相筛选、pH值与洗脱的优化等方面,综述了亲水作用色谱（HILIC）的方法开发策略,以及其在铂类抗癌药分析中的应用。