琥珀酸美托洛尔缓释微丸的制备及大鼠体内药代动力学

采用空白丸芯上药制备含药丸芯,以乙基纤维素为缓释材料,采用流化床包衣法制备了琥珀酸美托洛尔缓释微丸。以美国药典为标准对比,分析了自制品和市售品在pH 6.8 PBS、0.1 mol/L盐酸(pH 1.0)和水3种介质中的释放曲线,计算出相似因子(f2),并进一步对比分析了自制品和市售品在大鼠体内药代动力学行为差异。结果显示,采用60～88目空白丸芯,上药增重为100%,缓释层增重为22.7%～25.46%的缓释微丸与市售品在3种介质中的释放曲线的f2均大于50,相似性良好,同时方差分析显示两种制剂在大鼠体内的主要药代动力学参数没有显著性差异。     

LC-MS/MS法同时测定人血浆中盐酸二甲双胍和罗格列酮浓度及其药代动力学

目的:建立LC-MS/MS同时测定人血浆中盐酸二甲双胍及罗格列酮浓度的方法,测定志愿者单次和多次顿服二甲双胍罗格列酮胶囊后盐酸二甲双胍和罗格列酮的血药浓度,估算其药代动力学参数。方法:血浆中加入盐酸苯乙双胍为内标,经乙腈沉淀蛋白,取上清直接进行LC-MS/MS测定。色谱柱为Phenomenex Luna CN 100A(150 mm×2.0 mm ID),流动相为CH3OH-30 mmol/L NH4Ac,pH 5.0(80∶20),流速为0.2 mL/min;ESI-MS/MS选择性反应正离子检测。临床试验方案采用拉丁方试验设计。结果:盐酸二甲双胍和罗格列酮的血浆线性范围分别为5~3 000 ng/mL和1.5~500 ng/mL,检测限均为1ng/mL。测定了12名志愿者单次和多次顿服二甲双胍罗格列酮胶囊后的血药浓度经时过程。结论:本方法操作简单,专属性强,灵敏度高,准确性好。盐酸二甲双胍和罗格列酮体内动力学行为符合线性药物动力学特征。     

保心微丸中肉桂酸大鼠体内的药代动力学研究

目的研究保心微丸中肉桂酸在大鼠体内的药代动力学.方法采用高效液相色谱法测定肉桂酸血药浓度,色谱柱HypersilODSC18,150mm×4.6mm,5μm,柱温30C.流动相甲醇-1%醋酸(4555)流速0.5mL/min;检测波长273nm;进样量10μL.结果与结论建立了用高效液相色谱法测定肉桂酸血药浓度的方法,肉桂酸的峰面积(y)与浓度(X)之间的回归方程为y=4973.5348+42867.96678X,相关系数r=0.9998.肉桂酸在血浆中的回收率为97.5%,RSD为1.33%.检测限为0.15ng,大鼠血浆中最低检测浓度为75ng/mL.肉桂酸口服吸收在大鼠体内的药时过程为线性动力学过程,符合一级吸收一级消除的开放式室模型,t1/2(K.)为7.12min,tmax为53.29min,Cmax为0.20μg/mL,t1/2,2(Ke)为340.74min.该药代动力学参数可能是保心微丸中肉桂酸及有关成分等多种成分在大鼠体内的综合体现.     

关附甲素代谢产物关附胺醇在大鼠体内的药代动力学研究

目的:建立关附胺醇血药浓度的高效液相色谱 质谱联用分析方法,并探讨其在大鼠体内的药代动力学。方法:大鼠iv关附胺醇2 0mg/kg后不同时间点采血,利用LC- MS法测定血药浓度,并用3P87软件求算其药代动力学参数。结果:关附胺醇浓度在0 . 0 5～2 0 μg/mL范围内线性关系良好(r =0 . 9989)。绝对回收率大于80 % ,日内、日间RSD均小于15 %。大鼠iv关附胺醇2 0mg/kg后其主要动力学参数AUC、Vc、T1/2 、CLs分别为(5 . 5 9±1. 5 7)mg·h·L-1、(1 .19±0 .17)L·kg-1、(1 .88±0 . 2 4 )h、(3 .81±1. 0 2 )L·kg-1·h-1。排泄试验结果表明,给药2 4h后从尿中,胆汁中和粪便中排出的原型药物累计量分别相当于给药量的82 . 4 %、7. 5 %、4 . 7%。结论:该法专属性强,灵敏度高,可用于关附胺醇的体内定量分析。该药在大鼠体内消除较快,主要以原型从尿中排出。     

LC-MS/MS法测定大鼠血浆中芹黄春浓度及其在药代动力学研究中的应用

目的：研究大鼠分别灌胃和静脉给药芹黄春（芹菜素-7-O-β-D吡喃葡萄糖苷）溶液后,血浆中的芹黄春药物浓度及其药代动力学性质。方法6只雌性Sprague Dawley大鼠随机分为两组,每组3只,分别经灌胃(10 mg/kg)和静脉注射(2 mg/kg)给予芹黄春溶液后,按设计的时间点从大鼠眼内眦静脉丛采集血液样品,置于肝素化的离心管中,低温离心得血浆。采用甲醇沉淀法处理血浆生物样品,利用LC-MS/MS方法对血浆生物样品进行分析。药代动力学参数由Kinetica 2000软件进行计算。结果芹黄春浓度在1~2000 ng/mL范围内线性关系良好(r=0.9974),定量下限为1 ng/mL,低(8 ng/mL)、中(100 ng/mL)、高(1200 ng/mL)3个浓度的提取回收率均大于90%,日内日间精密度和稳定性的RSD均小于12.8%,日内日间准确度在91.5%~107%,方法学考察均符合要求。大鼠经静脉注射和灌胃芹黄春后,Cmax分别为(2363±96) ng/mL和(13.3±5.8) ng/mL,AUC0~∞分别为(796±201) h· ng/mL和(28.9±9.2) h· ng/mL,大鼠经静脉注射和后t1/2为(1.20±0.17) h,芹黄春的口服生物利用度约为0.6%。结论所建立的LC-MS/MS分析方法方便、快速,灵敏度高,准确度好,可用于大鼠血浆芹黄春浓度测定及其药代动力学的研究。     

LC-MS/MS法测定大鼠血浆中芍药内酯苷的浓度及其在毒代动力学研究中的应用

目的 建立测定大鼠血浆中芍药内酯苷的液相色谱-串联质谱(LC-MS/MS)法.方法 血浆样品经乙酸乙酯-异丙醇(95:5,V/V)提取后,以Poroshell 120 EC-C18柱(50 mm×2.1 mm,2.7μm)为分析柱,乙腈-水为流动相梯度洗脱,采用ESI源在多反应监测(MRM)方式下进行正离子检测.用于定量分析的离子反应为m/z 498.5→m/z 197.1(芍药内酯苷)和m/z 251.3→m/z 108.2(拉科酰胺,内标).结果 芍药内酯苷血浆浓度测定方法的线性范围为20~2000 ng/ml,日内、日间精密度RSD%均<15%,准确度(RE)在±8.06%之间.结论 该法适用于芍药内酯苷在大鼠体内的毒代动力学研究.     

布洛芬缓释胶囊的药代动力学研究

目的　研究布洛芬缓释胶囊单剂量及多剂量给药的药代动力学。方法　用HPLC法测定 1 0名受试者口服 60 0mg布洛芬缓释胶囊后的血药浓度 ,经 3P87药动学计算程序处理 ,得药动学参数。结果　单剂量口服后的Cmax为 (2 7.0 5± 4 .94) μg/ml、Tmax为 (4.4± 0 .70 )h、AUC为 (1 70 .51± 35 .89)h·μg/ml。多剂量口服后的Cmax为 (30 .66± 4 .67) μg/ml、Tmax为 (3 .7± 0 .67)h、AUC为 (1 76 .0 8± 1 8.88)h·μg/ml。波动系数为 1 .49。 结论　布洛芬缓释胶囊具有缓释效果     

HPLC-MS/MS法测定大鼠血浆中紫杉醇含量及其脂质体药代动力学研究

目的:建立一种测定大鼠血浆中紫杉醇浓度的高效液相色谱串联质谱（HPLC-MS/MS）分析方法,并研究其脂质体的药代动力学。方法:对6只SD大鼠静脉注射1 mg/kg的紫杉醇脂质体注射液,分别于给药后0、0.5、1、2、4、8、12、24、48 h自眼眶取血0.2 mL,C18反相色谱柱作为固定相,水（1 mmoL/L甲酸铵-0.05%甲酸）-甲醇作为流动相,ESI+扫描模式,MRM监测模式,药代动力学参数采用DAS 2.0软件进行计算。结果:紫杉醇在1～5 000 ng/mL范围内线性较好（R2 =0.999 7）,定量下限（LLOQ）为1 ng/mL,日内和日间精密度均低于10%,紫杉醇脂质体给药后血药浓度-时间曲线满足二室模型,主要药代动力学参数t1/2、Cmax、AUC0→t分别为（4.79±0.85）h、（49 785.12±1 618.45）μg/L、（38 753.08±6 144.66）μg/L·h。结论:本方法专属性和灵敏度高,结果可靠,能满足测定大鼠血浆中紫杉醇的定量需求。     

癌源性免疫球蛋白G全分子相互作用蛋白质的质谱鉴定及其功能分析

目的鉴定与癌源性免疫球蛋白G（IgG）全分子相互作用的蛋白质,为研究其生物功能提供重要的线索。方法分别用兔
抗人IgG全分子抗体和兔IgG在宫颈癌细胞系HeLa中进行免疫共沉淀,将得到的免疫复合物经电泳分离后进行银染。经比较
分析后3条差异条带切下来做质谱鉴定,质谱数据经Swiss-Prot数据库分析。最后对所得的蛋白质进行筛选和功能注释分析。
结果最终得到6个可能与癌源性IgG全分子相互作用的蛋白质。结论通过免疫共沉淀结合质谱分析的方法最终得到了6个
可能与癌源性IgG全分子相互作用的蛋白质,为其功能研究提供了重要线索。
     

苯磺酸左旋氨氯地平血药浓度的LC-MS/MS法测定及其人体内手性转化可能性考察

目的 建立LC-MS/MS法测定人血浆中左旋氨氯地平血药浓度并进行体内手性转化可能性考察。 方法 以氯氮为内标, 采用CHIRAL-AGP柱(150.0 mm×4.0 mm, 5 μm)对氨氯地平消旋体进行分离手性对映体, 流动相为10 mmol/L乙酸铵缓冲液(pH 4.38)-异丙醇(982, V/V);选择固相萃取法提取10例健康男性受试者单次口服苯磺酸左旋氨氯地平片2.5 mg 后的血浆样品, 大气压化学电离源(APCI)结合正离子MRM扫描分析测定人体内S-(-)-氨氯地平浓度, 其中氨氯地平和内标离子对分别是m/z 409.0→237.9和m/z 300.0→282.0。 结果 S-(-)-/R-(+)-氨氯地平对映体血药浓度在0.103 1~20.62 μg/L范围内线性关系良好(r=0.999 8, r=0.999 7), 绝对回收率大于70.0%, 相对回收率均在85.0%~115.0% 范围内, 日内和日间RSD均小于15.0%。10例健康男性受试者单剂量口服苯磺酸左旋氨氯地平片2.5 mg后体内不同时间点血浆样品均未检测到R-(+)-氨氯地平对映体, S-(-)-氨氯地平在健康男性受试者体内的药代动力学参数t1/2为(42.77±8.08) h, Cmax为(3.06±0.51) μg/L, tmax为(6.3±1.0) h, MRT为(69.25±8.04) h, AUC0-144为(176.20±31.89) h·μg·L-1, AUC0-∞为(197.92±37.54) h·μg·L-1。 结论 本方法 选择性强, 灵敏度高, 无杂质干扰, 精密度好, 成功地应用于苯磺酸左旋氨氯地平人体药代动力学分析, 并证实S-(-)-氨氯地平对映体在健康男性受试者体内未发现其手性转化。     

LC-MS/MS method for determination of megestrol in human plasma and its application in bioequivalence study

A rapid and highly selective liquid chromatography-tandem mass spectrometric （LC-MS/MS） method for the determination of megestrol in human plasma was described using medry- sone as internal standard （IS）. Blood samples were collected from 20 healthy volunteers after oral ad- ministration of 160 mg megestrol acetate dispersible tablets. The analytes were extracted by liq- uid-liquid extraction procedure and separated on a hanbon lichrospher column with the mobile phase of methanol and water containing 0.1% formic acid and 20 mmol/L ammonium acetate （5：1, v/v）. Positive ion electrospray ionization with multiple reaction-monitoring mode （MRM） was employed by monitor- ing the transitions m/z 385.5-325.4 and m/z 387.5-327.4 for megestrol and medrysone, respectively. Under the isocratic separation conditions, the chromatographic rim time was approximately 2.54 min for megestrol and 2.59 min for medrysone. The calibration curve range was from 0.5 to 200.0 ng/mL. The inter-batch and intra-batch precision and accuracy were less than 5.2% relative standard deviation （RSD） and 6.4% relative error （RE）. The proposed method was successfully applied in the bioequivalence study of megestrol acetate dispersible tablets.     

血中22种常见毒/药物的液相色谱-串联质谱法快速鉴定

目的 为了快速鉴定血中22种常见毒/药物.方法 采用液相色谱-串联质谱作为分离测试手段,检测模式为多反应监测模式,血液1 mL中加入2 mL乙酸乙酯,涡旋震荡3min,5000 r/min离心5 min,取有机层,重复萃取过程三次,合并有机相,氮吹至干,残余物中加入100 L流动相,涡旋震荡1 min,取5 L进样鉴定.结果 10 min内完成22种常见毒药物的快速鉴定,检出限为0.03到6.00ng/mL范同内;在中国合格评定委员会组织的能力验证考核中,采用方法所测结果满意.结论 所开发方法开速、准确、灵敏.

Abstract：

Objective To develop a method for rapid identification of 22 abused drugs and organophosphorus pesticides in the blood. Methods Liquid chromatography-tandem mass spectrometry (LC-MS/ MS) in multiple-reaction monitoring mode (MRM) was employed for detecting the drugs and pesticides in the blood. The MRM database and criteria for identification were established, and ethyl acetate was used for extraction of the drugs. After 3 rounds of extractions of the blood sample (1 mL) using 2 mL ethyl acetate, the extract was vortexed for 3 min and centrifuged at 5000 r/min. Each organic phase was combined and evaporated by gentle N2 gas. The residue was re-dissolved in 100 L mobile phase, from which 5 L was taken for LC-MS/MS detection. Results The detection of the 22 target compounds could be completed within 10 min. The limit of detection of the target compound ranged from 0.03 to 6.00 ng/ml. Satisfactory results were obtained in proficiency testing program organized by China National Accreditation Service for Conformity Assessment. Conclusion The method we established is rapid, selective and sensitive for detecting the 22 abused drugs and organophosphorus pesticides.     

苯扎贝特缓释微丸胶囊的研制及体内外评价

目的:研制苯扎贝特缓释微丸胶囊。方法:采用离心造粒技术制备苯扎贝特缓释微丸胶囊;以体外释药和犬的药代动力学参数进行评价。结果:微丸得率86.5%,载药量87.8%;体外释放曲线符合一级方程,相似因子f2为74.23;自制缓释微丸胶囊单剂量犬口服给药后的tmax(3.7±0.5)h,cmax(36.36±0.98)μg/mL,AUC0-∞(337.83±7.29)μg.h/mL;苯扎贝特缓释片(参比片)的tmax(4.1±0.4)h,cmax(39.66±0.94)μg/mL,AUC0-∞(327.27±8.97)μg.h/mL。结论:以进口的缓释片为参比片,自制的苯扎贝特缓释微丸胶囊具有良好的缓释作用;相对生物利用度为(103.3±2.8)%,两种制剂生物等效。     

An LC-MS/MS Method for the Simultaneous Determination of Lycorine and Galanthamine in Rat Plasma and Its Application to Pharmacokinetic Study of Lycoris Radiata Extract in Rats

A rapid, sensitive, and selective liquid chromatography-tandem mass spectrometry was developed for the simultaneous determination of lycorine and galanthamine, two major constituents in Lycoris radiata extract, in rat plasma. Liquid-liquid extraction with ethyl ether was carried out using diphenhydramine as the internal standard The two bioactive alkaloids were separated on a Zorbax SB-C18 reserved-phase column （150 mm× 4.6 mm, i.d., 5 μm） by gradient elution using a mobile phase consisting of methanol with 0.1% formic acid （A） and water with 0.1% formic acid （B） at a flow rate of 0.6 mL/min. All analytes showed good linearity over a wide concentration range （r^2〉0.99） and the lower limit of quantification was 3.00 ng/mL for each analyte. The average extraction re- covery of the analytes from rat plasma was more than 82.15%, and the intra-day and inter-day accuracy and preci- sion of the assay were less than 12.6%. The validated method was successfully applied to monitoring the concen- trations and pharmacokinetic studies of two Amaryllidaceous alkaloids in rat plasma after an oral administration of Lycoris radiata extract.     

Influence of traditional Chinese medicines on the in vivo metabolism of lopinavir/ritonavir based on UHPLC-MS/MS analysis

A fast, reliable, and cost-effective liquid chromatography-tandem mass spectrometry method was established to determine the effects of the traditional Chinese medicine employed to treat coronavirus disease 2019, namely, Lianhua Qingwen granules, Huoxiang Zhengqi capsules, Jinhua Qinggan granules, Shufeng Jiedu capsules, and Angong Niuhuang pills, on the pharmacokinetics of lopinavir/ritonavir in rats. Blood samples were prepared using the protein precipitation method and atazanavir was selected as the internal standard (IS). Separation was performed on an Agilent ZORBAX eclipse plus C₁₈ (2.1 mm × 50 mm, 1.8 μm) column using acetonitrile and water containing 0.1% formic acid as the mobile phase for gradient elution. The flow rate was 0.4 mL/min and the injection volume was 2 μL. Agilent Jet Stream electrospray ionization was used for mass spectrometry detection under positive ion multiple reaction monitoring mode at a transition of m/z 629.3→447.3 for lopinavir, m/z 721.3→296.1 for ritonavir, and m/z 705.4→168.1 for the IS. The method showed good linearity in the concentration range of 25–2500 ng/mL (r=0.9981) for lopinavir and 5–500 ng/mL (r=0.9984) for ritonavir. The intra-day and inter-day precision and accuracy were both within ±15%. Items, such as dilution reliability and residual effect, were also within the acceptable limits. The method was used to determine the effects of five types of traditional Chinese medicines on the pharmacokinetics of lopinavir/ritonavir in rats. The pharmacokinetic results showed that the half-life of ritonavir in the groups administered Lianhua Qingwen granules and Huoxiang Zhengqi capsules combined with lopinavir/ritonavir was prolonged by approximately 1.5- to 2-fold relative to that in the control group. Similarly, the pharmacokinetic parameters of lopinavir were altered. Overall, the results of this study offer important theoretical parameters for the effective clinical use of five types of traditional Chinese medicines combined with lopinavir/ritonavir to reduce the occurrence of clinical adverse reactions.     

Influence of traditional Chinese medicines on the in vivo metabolism of lopinavir/ritonavir based on UHPLC-MS/MS analysis

A fast, reliable, and cost-effective liquid chromatography-tandem mass spectrometry method was established to determine the effects of the traditional Chinese medicine employed to treat coronavirus disease 2019, namely, Lianhua Qingwen granules, Huoxiang Zhengqi capsules, Jinhua Qinggan granules, Shufeng Jiedu capsules, and Angong Niuhuang pills, on the pharmacokinetics of lopinavir/ritonavir in rats. Blood samples were prepared using the protein precipitation method and atazanavir was selected as the internal standard (IS). Separation was performed on an Agilent ZORBAX eclipse plus C₁₈ (2.1 mm × 50 mm, 1.8 μm) column using acetonitrile and water containing 0.1% formic acid as the mobile phase for gradient elution. The flow rate was 0.4 mL/min and the injection volume was 2 μL. Agilent Jet Stream electrospray ionization was used for mass spectrometry detection under positive ion multiple reaction monitoring mode at a transition of m/z 629.3→447.3 for lopinavir, m/z 721.3→296.1 for ritonavir, and m/z 705.4→168.1 for the IS. The method showed good linearity in the concentration range of 25–2500 ng/mL (r=0.9981) for lopinavir and 5–500 ng/mL (r=0.9984) for ritonavir. The intra-day and inter-day precision and accuracy were both within ±15%. Items, such as dilution reliability and residual effect, were also within the acceptable limits. The method was used to determine the effects of five types of traditional Chinese medicines on the pharmacokinetics of lopinavir/ritonavir in rats. The pharmacokinetic results showed that the half-life of ritonavir in the groups administered Lianhua Qingwen granules and Huoxiang Zhengqi capsules combined with lopinavir/ritonavir was prolonged by approximately 1.5- to 2-fold relative to that in the control group. Similarly, the pharmacokinetic parameters of lopinavir were altered. Overall, the results of this study offer important theoretical parameters for the effective clinical use of five types of traditional Chinese medicines combined with lopinavir/ritonavir to reduce the occurrence of clinical adverse reactions.     

HPLC-TOF/MS对中药复方扶正平消胶囊化学成分的鉴别

目的 运用高效液相-高分辨飞行时间质谱(HPLC-TOF/MS)技术对中药复方扶正平消胶囊化学成分进行鉴别。方法 色谱分离采用Agilent Eclipse plus C18 (4.6 mm×250 mm,5 μm) 色谱柱;流动相为乙腈和0.1％甲酸水溶液,梯度洗脱:0~90 min,5%A~95%A;柱温25℃;流速1 ml/min,柱后分流比为31。质谱定性采用飞行时间质谱,电喷雾离子源(ESI),正负离子模式共同监测,质量数扫描范围m/z 100~1 100。结果 共鉴别出扶正平消胶囊中247种的化学成分,其中正离子模式下168个、负离子模式下103个、正负离子均有响应24个,并对成分进行了药材归属。结论 建立了一种基于HPLC-TOF/MS技术对中药复方扶正平消胶囊中的化学成分进行鉴别的有效方法,为其质量控制及体内的深入研究奠定了基础。