目的建立定量测定大鼠血浆中顺式二苯乙烯苷[cis-2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷(TSG)]与反式二苯乙烯苷(trans-TSG)血药浓度的UPLC-MS/MS方法,系统探究2种异构体在大鼠体内的药动学差异。方法将40只雄性SD大鼠随机分为...目的建立定量测定大鼠血浆中顺式二苯乙烯苷[cis-2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷(TSG)]与反式二苯乙烯苷(trans-TSG)血药浓度的UPLC-MS/MS方法,系统探究2种异构体在大鼠体内的药动学差异。方法将40只雄性SD大鼠随机分为4组(每组10只),即cis-TSG ig给药组、cis-TSG尾iv给药组、trans-TSG ig给药组、transTSG尾iv给药组。ig组给药剂量为100 mg·kg^(-1),尾iv组给药剂量为10 mg·kg^(-1);于给药前(0 h)及给药后0.030、0.083、0.160、0.250、0.330、0.500、1.000、2.000、4.000、6.000、8.000、10.000、12.000、24.000 h,通过眼静脉丛采集血样。以虎杖苷为内标,测定上述2个成分在不同时间点的血药浓度;利用DAS 1.0软件计算药动学参数,分析其药动学行为差异。结果所建立的UPLC-MS/MS方法在专属性、稳定性、准确度与精密度方面均符合体内药物浓度测定的技术要求,且无明显基质效应、提取回收率稳定。药动学结果显示:①给药途径对暴露水平影响显著:2种成分经iv给药后的血药浓度-时间曲线下面积(AUC)、达峰浓度(C_(max))均显著高于ig给药,且达峰时间(t_(max))更短,提示iv给药可使药物直接、快速进入体循环;②异构体间暴露差异显著:无论是iv还是ig给药,cis-TSG的AUC、C_(max)均显著高于trans-TSG(P<0.05),cis-TSG的消除半衰期(t_(1/2))长于trans-TSG[iv(7.03 h vs 2.63 h)、ig(16.90 h vs 8.46 h)],提示2种异构体在吸收、代谢或分布过程中存在本质差异;③口服生物利用度较低:cis-TSG与trans-TSG的口服绝对生物利用度分别为28.46%、27.35%。结论cis-TSG与trans-TSG在大鼠体内的药动学行为存在显著差异,主要体现为cis-TSG的体内暴露水平更高、消除更缓慢;且2种成分的口服生物利用度均较低。展开更多
印章印文作为现代社会中文件合同的重要标识物,其形成时间的鉴定是文书鉴定领域关注的重点和难点之一。采用溶剂提取法结合超高效液相色谱-串联质谱(Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry,UPLC-MS/MS...印章印文作为现代社会中文件合同的重要标识物,其形成时间的鉴定是文书鉴定领域关注的重点和难点之一。采用溶剂提取法结合超高效液相色谱-串联质谱(Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry,UPLC-MS/MS)技术对14种印油、印泥和印台印文的主要染料成分含量进行测定分析,以探究目标染料含量随时间的变化规律。通过单因素实验结合XGBoost机器学习算法对前处理条件进行优化,确定了最佳前处理条件,包括提取剂种类(DMF∶乙腈=2∶1)、样品量(0.1 cm×1.0 cm)、提取剂剂量(1000μL)和提取方式(振荡30 s与超声5 min)。利用UPLC-MS/MS对金光红、金光红C两种目标染料进行检测,在10~500μg/L范围内线性良好(r^(2)为0.9949~0.9996),回收率分别在100.49%~102.17%与98.30%~99.30%之间,相对标准偏差为0.53%~4.65%。研究结果表明,大部分印文中染料的检出量随时间推移呈现递减规律,且一萃和二萃染料检出量比值也表现出明显递减趋势。所建立的UPLC-MS/MS法可相对明确地区分24个月内不同时间段的印章印文,为印文形成时间的鉴定提供了科学依据与可行的技术方案。展开更多
Objectives:Acquired resistance to paclitaxel represents a critical barrier to the effective chemotherapy of non-small cell lung cancer(NSCLC).The present study aimed to elucidate the molecular and pharmacological mech...Objectives:Acquired resistance to paclitaxel represents a critical barrier to the effective chemotherapy of non-small cell lung cancer(NSCLC).The present study aimed to elucidate the molecular and pharmacological mechanisms promoting paclitaxel resistance in NSCLC and to explore potential strategies for overcoming this resistance.Methods:Here,we report an integrated pharmacological and analytical approach to quantify paclitaxel disposition and overcome resistance in a A549/TAX cell model(paclitaxel-resistant A549 cells).Results:Cell counting kit-8(CCK-8)assay,colony formation,and apoptosis assays confirmed that A549/TAX cells exhibited marked resistance to paclitaxel relative to parental A549 cells.Based on transcriptome profiling by RNA sequencing analysis and validation by western blotting assay,we found that the expression of the ATP-binding cassette subfamily B member 1(ABCB1)(the encoded protein is termed P-glycoprotein)was significantly upregulated in resistant cells.By using ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS),we demonstrated that ABCB1 overexpression promotes enhanced efflux of intracellular paclitaxel,thereby lowering its cytotoxic accumulation.Genetic silencing of ABCB1 or pharmacological inhibition with the specific P-glycoprotein modulator elacridar or tariquidar restored intracellular paclitaxel levels,as determined by UPLC-MS/MS,and synergistically decreased cell viability as observed in CCK-8 assay.Conclusion:These findings reveal that the ABCB1-mediated drug efflux is a crucial mechanism underlying paclitaxel resistance in NSCLC cells,with UPLC-MS/MS serving as a sensitive analytical method to detect paclitaxel concentration.Inhibition of ABCB1 is a promising therapeutic strategy to resensitize resistant tumor cells to paclitaxel.展开更多
文摘目的建立定量测定大鼠血浆中顺式二苯乙烯苷[cis-2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷(TSG)]与反式二苯乙烯苷(trans-TSG)血药浓度的UPLC-MS/MS方法,系统探究2种异构体在大鼠体内的药动学差异。方法将40只雄性SD大鼠随机分为4组(每组10只),即cis-TSG ig给药组、cis-TSG尾iv给药组、trans-TSG ig给药组、transTSG尾iv给药组。ig组给药剂量为100 mg·kg^(-1),尾iv组给药剂量为10 mg·kg^(-1);于给药前(0 h)及给药后0.030、0.083、0.160、0.250、0.330、0.500、1.000、2.000、4.000、6.000、8.000、10.000、12.000、24.000 h,通过眼静脉丛采集血样。以虎杖苷为内标,测定上述2个成分在不同时间点的血药浓度;利用DAS 1.0软件计算药动学参数,分析其药动学行为差异。结果所建立的UPLC-MS/MS方法在专属性、稳定性、准确度与精密度方面均符合体内药物浓度测定的技术要求,且无明显基质效应、提取回收率稳定。药动学结果显示:①给药途径对暴露水平影响显著:2种成分经iv给药后的血药浓度-时间曲线下面积(AUC)、达峰浓度(C_(max))均显著高于ig给药,且达峰时间(t_(max))更短,提示iv给药可使药物直接、快速进入体循环;②异构体间暴露差异显著:无论是iv还是ig给药,cis-TSG的AUC、C_(max)均显著高于trans-TSG(P<0.05),cis-TSG的消除半衰期(t_(1/2))长于trans-TSG[iv(7.03 h vs 2.63 h)、ig(16.90 h vs 8.46 h)],提示2种异构体在吸收、代谢或分布过程中存在本质差异;③口服生物利用度较低:cis-TSG与trans-TSG的口服绝对生物利用度分别为28.46%、27.35%。结论cis-TSG与trans-TSG在大鼠体内的药动学行为存在显著差异,主要体现为cis-TSG的体内暴露水平更高、消除更缓慢;且2种成分的口服生物利用度均较低。
基金supported by grants from the National Natural Science Foundation of China(Grant No.82172840)Gusu Health Talents Project of Suzhou Municipal Health Commission(Grant Nos.GSWS2023007 and GSWS2022062)+2 种基金Suzhou Science and Technology Development Plan Project(Grant No.SYW2024005)Chinese Pharmaceutical Association Hospital Pharmacy Department(Grant No.CPA-Z05-ZC-2024002)Jiangsu Research Hospital Association for Precision Medication(Grant No.JY202202).
文摘Objectives:Acquired resistance to paclitaxel represents a critical barrier to the effective chemotherapy of non-small cell lung cancer(NSCLC).The present study aimed to elucidate the molecular and pharmacological mechanisms promoting paclitaxel resistance in NSCLC and to explore potential strategies for overcoming this resistance.Methods:Here,we report an integrated pharmacological and analytical approach to quantify paclitaxel disposition and overcome resistance in a A549/TAX cell model(paclitaxel-resistant A549 cells).Results:Cell counting kit-8(CCK-8)assay,colony formation,and apoptosis assays confirmed that A549/TAX cells exhibited marked resistance to paclitaxel relative to parental A549 cells.Based on transcriptome profiling by RNA sequencing analysis and validation by western blotting assay,we found that the expression of the ATP-binding cassette subfamily B member 1(ABCB1)(the encoded protein is termed P-glycoprotein)was significantly upregulated in resistant cells.By using ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS),we demonstrated that ABCB1 overexpression promotes enhanced efflux of intracellular paclitaxel,thereby lowering its cytotoxic accumulation.Genetic silencing of ABCB1 or pharmacological inhibition with the specific P-glycoprotein modulator elacridar or tariquidar restored intracellular paclitaxel levels,as determined by UPLC-MS/MS,and synergistically decreased cell viability as observed in CCK-8 assay.Conclusion:These findings reveal that the ABCB1-mediated drug efflux is a crucial mechanism underlying paclitaxel resistance in NSCLC cells,with UPLC-MS/MS serving as a sensitive analytical method to detect paclitaxel concentration.Inhibition of ABCB1 is a promising therapeutic strategy to resensitize resistant tumor cells to paclitaxel.
