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Push forward LC-MS-based therapeutic drug monitoring and pharmacometabolomics for anti-tuberculosis precision dosing and comprehensive clinical management

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Journal of Pharmaceutical Analysis 2024年第1期14卷 16-38页

作者： Nguyen Quang Thu Nguyen Tran Nam Tien Nguyen Thi Hai Yen Thuc-Huy Duong Nguyen Phuoc Long Huy Truong Nguyen Department of Pharmacology and PharmacoGenomics Research Center Inje University College of MedicineBusan47392Republic of Korea Department of Chemistry University of EducationHo Chi Minh City700000Viet Nam Faculty of Pharmacy Ton Duc Thang UniversityHo Chi Minh City700000Viet Nam

The spread of tuberculosis(TB),especially multidrug-resistant TB and extensively drug-resistant TB,has strongly motivated the research and development of new anti-TB drugs.New strategies to facilitate drug combinations,including pharmacokinetics-guided dose optimization and toxicology studies of first-and second-line anti-TB drugs have also been introduced and recommended.Liquid chromatography-mass spectrometry(LC-MS)has arguably become the gold standard in the analysis of both endo-and exo-genous compounds.This technique has been applied successfully not only for therapeutic drug monitoring(TDM)but also for pharmacometabolomics analysis.TDM improves the effectiveness of treatment,reduces adverse drug reactions,and the likelihood of drug resistance development in TB patients by determining dosage regimens that produce concentrations within the therapeutic target window.Based on TDM,the dose would be optimized individually to achieve favorable outcomes.Pharmacometabolomics is essential in generating and validating hypotheses regarding the metabolism of anti-TB drugs,aiding in the discovery of potential biomarkers for TB diagnostics,treatment monitoring,and outcome evaluation.This article highlighted the current progresses in TDM of anti-TB drugs based on LC-MS bioassay in the last two decades.Besides,we discussed the advantages and disadvantages of this technique in practical use.The pressing need for non-invasive sampling approaches and stability studies of anti-TB drugs was highlighted.Lastly,we provided perspectives on the prospects of combining LC-MS-based TDM and pharmacometabolomics with other advanced strategies(pharmacometrics,drug and vaccine developments,machine learning/artificial intelligence,among others)to encapsulate in an all-inclusive approach to improve treatment outcomes of TB patients.

关键词： Tuberculosis Therapeutic drug monitoring LC-MS MIPD Pharmacometabolomics Precision medicine

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A widely targeted metabolite modificomics strategy for modified metabolites identification in tomato

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Journal of Integrative Plant Biology 2024年第4期66卷 810-823页

作者： Jun Yang Ridong Chen Chao Wang Chun Li Weizhen Ye Zhonghui Zhang Shouchuang Wang School of Breeding and Multiplication(Sanya Institute of Breeding and Multiplication) Hainan UniversitySanya 572025China School of Tropical Agriculture and Forestry Hainan UniversityHaikou 572208China

The structural and functional diversity of plant metabolites is largely created via chemical modification of a basic backbone.However,metabolite modifications in plants have still not been thoroughly investigated by metabolomics approaches.In this study,a widely targeted metabolite modificomics(WTMM)strategy was developed based on ultra-high performance liquid chromatography-quadrupole-linear ion trap(UHPlc-Q-Trap)and UHPlc-Q-Exactive-Orbitrap(UHPlc-QE-Orbitrap),which greatly improved the detection sensitivity and the efficiency of identification of modified metabolites.A metabolite modificomics study was carried out using tomato as a model,and over 34,000 signals with ms2 information were obtained from approximately 232 neutral loss transitions.Unbiased metabolite profiling was also performed by utilizing high-resolution mass spectrometry data to annotate a total of 2,118 metabolites with 125 modification types;of these,165 modified metabolites were identified in this study.Next,the WTMM database was used to assess diseased tomato tissues and 29 biomarkers were analyzed.In summary,the WTMM strategy is not only capable of large-scale detection and quantitative analysis of plant-modified metabolites in plants,but also can be used for plant biomarker development.

关键词： LC-MS metabolic diversity metabolite modificomics modified groups tomato

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LC-MS测定大鼠脑脊液与血浆中西达本胺浓度及其药动学研究

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中国现代应用药学 2023年第3期40卷 342-350页

作者：闫珍顾利强徐晓珍舒艳陈灵芳史煜华张丽丽陈浩张城达张升王佳虹张立将杭州医学院安全性评价研究中心浙江省药物安全性评价技术研究重点实验室杭州310053

目的建立快速检测SD大鼠脑脊液和血浆中西达本胺浓度的LC-MS测定方法,并应用于大鼠静脉注射西达本胺后的脑脊液和血浆中药动学研究。方法对建立的检测大鼠脑脊液和血浆中西达本胺浓度的LC-MS分析方法进行专属性、精密度、准确性、基质... 详细信息

目的建立快速检测SD大鼠脑脊液和血浆中西达本胺浓度的LC-MS测定方法,并应用于大鼠静脉注射西达本胺后的脑脊液和血浆中药动学研究。方法对建立的检测大鼠脑脊液和血浆中西达本胺浓度的LC-MS分析方法进行专属性、精密度、准确性、基质效应及稳定性等考察验证。♂SD大鼠12只,分为2组,每组6只,分别静脉注射给予西达本胺3,6 mg·kg^(-1)剂量,于给药后0.083,0.25,0.5,1,1.5,2,3 h采集脑脊液和血浆,采用LC-MS测定大鼠脑脊液和血浆中西达本胺的浓度变化,并利用DAS药动学软件拟合脑脊液和血浆中AUC、C_(max)、T_(max)、t_(1/2)等主要药动学参数,计算血脑屏障透过率。结果方法学研究显示,大鼠脑脊液中西达本胺线性范围为0.2~64 ng·mL^(-1)(r=0.999 0,定量下限为0.2 ng·mL^(-1)),血浆中西达本胺的线性范围为2~1 000 ng·mL^(-1)(r=0.999 1,定量下限为2 ng·mL^(-1)),脑脊液和血浆中西达本胺的日内精密度、日间精密度、准确度均符合方法学验证要求。大鼠药动学研究显示,大鼠静脉注射给予西达本胺3,6 mg·kg^(-1)剂量后,脑脊液中的AUC_(0-t)分别为16.01,32.09 ng^(-1)·mL^(-1)·h,C_(max)分别为17.58,28.37 ng·mL^(-1),t_(1/2)分别为1.14,1.23 h;血浆的AUC_(0-t)分别为2 570.11,4 537.63 ng^(-1)·mL^(-1)·h,C_(max)分别为4 366.54,9 103.97 ng·mL^(-1),t_(1/2)分别为0.83,0.77 h。2组西达本胺的平均血脑屏障透过率分别为0.62%和0.72%。结论本研究建立了简便、迅速、灵敏的检测大鼠脑脊液和血浆中西达本胺浓度的分析方法,并成功应用于大鼠药动学研究,静脉注射西达本胺能够有效地透过血脑屏障,可以实现直接作用于脑部肿瘤的目的。

关键词：西达本胺 LC-MS 血浆脑脊液药动学

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氯化钠诱导白木香悬浮细胞合成苯乙基色酮类成分的LC-MS分析

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中国中药杂志 2023年第9期48卷 2480-2489页

作者：杜宇张晓雪张泽坤王雯静张蓓蓓张明亮王阳葛祥宇史社坡北京中医药大学中药学院、中药现代研究中心北京100029

采用超高效液相色谱-串联四极杆静电场轨道阱质谱(UPlc-Q-Exactive-ms)结合超高效液相色谱-串联三重四极杆质谱(UPlc-QQQ-ms/ms)对氯化钠诱导的白木香悬浮细胞中苯乙基色酮类成分进行定性和定量分析。UPlc-Q-Exactive-ms和UPlc-QQQ-ms/M... 详细信息

采用超高效液相色谱-串联四极杆静电场轨道阱质谱(UPlc-Q-Exactive-ms)结合超高效液相色谱-串联三重四极杆质谱(UPlc-QQQ-ms/ms)对氯化钠诱导的白木香悬浮细胞中苯乙基色酮类成分进行定性和定量分析。UPlc-Q-Exactive-ms和UPlc-QQQ-ms/ms均采用Waters T3色谱柱(2.1 mm×50 mm,1.8μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相分别进行梯度洗脱色谱分离,质谱分析均采用电喷雾正离子模式采集。利用UPlc-Q-Exactive-ms从氯化钠诱导的白木香悬浮细胞样品中鉴定出苯乙基色酮类化合物47个,其中简单苯乙基色酮及其苷22个、四羟基四氢苯乙基色酮10个、三元氧环四氢苯乙基色酮15个。通过UPlc-QQQ-ms/ms技术对氯化钠诱导处理后白木香悬浮细胞中25个苯乙基色酮进行了定量。通过2种液质联用技术快速、高效地对氯化钠诱导白木香悬浮细胞中苯乙基色酮类成分进行定性、定量分析,为利用体外培养等生物技术制备沉香中主要的药效和香味物质苯乙基色酮提供重要参考。

关键词：白木香悬浮细胞沉香苯乙基色酮 LC-MS

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DMMIC derivatization-assisted liquid chromatography-mass spectrometry method for metabolite profiling of the glutathione anabolic pathway in esophageal cancer tissues and cells

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Journal of Pharmaceutical Analysis 2023年第11期13卷 1365-1373页

作者： Li Liu Yu-Han Lu Min-Dan Wang Qun-Fei Zhao Xiu-Ping Chen Hang Yin Chen-Guo Feng Fang Zhang The Research Center of Chiral Drugs Shanghai Frontiers Science Center for TCM Chemical BiologyInnovation Research Institute of Traditional Chinese MedicineShanghai University of Traditional Chinese MedicineShanghai201203China Department of Thoracic Surgery Renji HospitalSchool of MedicineShanghai Jiaotong UniversityShanghai200127China School of Public Health Shanghai University of Traditional Chinese MedicineShanghai201203China

In this work,a new pyrylium derivatization-assisted liquid chromatography-mass spectrometry(LC-MS)method was developed for metabolite profiling of the glutathione anabolic pathway(GAP)in cancer tissues and cells.The pyrylium salt of 6,7-dimethoxy-3-methyl isochromenylium tetrafluoroborate(DMMIC)was used to label the amino group of metabolites,and a reductant of dithiothreitol(DTT)was employed to stabilize the thiol group.By combining DMMIC derivatization with LC-MS,it was feasible to quantify the 13 main metabolites on the GAP in complex biological samples,which had good linearity(R^(2)=0.99810.9999),precision(interday precision of 1.6%e19.0%and intraday precision of 1.4%e19.8%)and accuracy(83.4%-115.7%).Moreover,the recovery assessments in tissues(82.5%e107.3%)and in cells(98.1%e118.9%)with GSH-^(13)C2,^(15)N,and Cys-^(15)N demonstrated the reliability of the method in detecting tissues and cells.Following a methodological evaluation,the method was applied successfully to investigate difference in the GAP between the carcinoma and para-carcinoma tissues of esophageal squamous cell carcinoma(ESCC)and the effect of p-hydroxycinnamaldehyde(CmsP)on the GAP in KYSE150 esophageal cancer cells.The results demonstrate that the developed method provides a promising new tool to elucidate the roles of GAP in physiological and pathological processes,which can contribute to research on drugs and diseases.

关键词： Glutathione anabolic pathway Metabolite profiling DMMIC derivatization LC-MS Esophageal squamous cell carcinoma p-Hydroxycinnamaldehyde KYSE-150cell

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山银花油GC-ms及LC-MS的全成分表征及体外抗炎和抗氧化活性研究

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天然产物研究与开发 2023年第11期35卷 1842-1856页

作者：刘恃君陆玉爱李利汤杨黔南林丽美谢明霞夏伯候湖南中医药大学药学院湘产大宗药材品质评价湖南省重点实验室长沙410208

山银花是一种有价值的药用资源,但其油的化学成分和生物活性尚未深入研究。本文通过气相色谱-质谱仪联用法(GC-ms)及超高效液相色谱-高分辨质谱联用(UHPlc-QE-ms)对超临界CO_(2)萃取法得到的山银花油进行系统的化学成分表征,并探讨其体... 详细信息

山银花是一种有价值的药用资源,但其油的化学成分和生物活性尚未深入研究。本文通过气相色谱-质谱仪联用法(GC-ms)及超高效液相色谱-高分辨质谱联用(UHPlc-QE-ms)对超临界CO_(2)萃取法得到的山银花油进行系统的化学成分表征,并探讨其体外抗氧化及细胞抗炎能力等生物活性。GC-ms共鉴别出66种化合物,主要为脂肪酸类、甾醇类和烷烃类化合物。UHPlc-QE-ms在正负离子模式下分别鉴别出83和71种化合物,主要为萜类、黄酮类等化合物。抗氧化结果表明山银花油清除ABTS自由基能力呈良好的剂量依赖性,DPPH自由基的IC_(50)值为3.49 mg/mL。同时抗炎实验表明山银花油可显著抑制脂多糖(LPS)诱导小鼠巨噬细胞(RAW 264.7)炎性因子IL-6(P<0.05)和NO(P<0.05)的释放,发挥抗炎作用。以上结果表明,山银花油化学成分复杂多样,是一种具有较优抗炎作用的天然活性来源,且在食品医药中有较好的开发和应用价值。

关键词：山银花挥发油 GC-ms LC-MS 生物活性

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Expression and Characterization of a Novelλ-Carrageenase Cgl150A_Wa from Wenyingzhuangia aestuarii

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Journal of Ocean University of China 2024年第1期23卷 209-215页

作者： SUN Yuhao CAO Siqi ZHANG Yuying XUE Changhu XIAO Hang CHANG Yaoguang College of Food Science and Engineering Ocean University of ChinaQingdao 266003China Marine Life Research Center Laoshan National LaboratoryQingdao 266237China Department of Food Science University of Massachusetts AmherstMA 01003United States

λ-Carrageenan is a highly sulfated polysaccharide alternating of 1,4-O-α-D-galactopyranose-2,6-sulfate(D2S,6S)and 1,3-O-β-D-galactopyranose-2-sulfate(G2S).λ-Carrageenases are desirable tools forλ-carrageenan degradation.Based on the genome mining,a novelλ-carrageenase Cgl150A_Wa was cloned from the bacterium Wenyingzhuangia aestuarii and expressed in Escherichia coli.Cgl150A_Wa was an endo-acting enzyme and exhibited its maximum activity at 30℃and pH 8.0.By employing a glycomics strategy that combined ultra-performance liquid chromatography-mass spectrometry analysis and glycoinformatics,Cgl150A_Wa was proven to degradeλ-carrageenan octaose and hexaose,and the major hydrolysis product of Cgl150A_Wa wasλ-carrageenan tetrose.In addition to the typicalλ-carrageenan motifs,the active center of Cgl150A_Wa might tolerate desulfatedλ-carrageenan motifs.Cgl150A_Wa is a potential biotechnological tool for preparingλ-carrageenan oligosaccharides and structural investigation.

关键词： carrageenan λ-carrageenase LC-MS oligosaccharide GH150

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Analysis of human brain tissue derived from DBS surgery

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Translational Neurodegeneration 2022年第1期11卷 625-640页

作者： Salla M.Kangas Jaakko Teppo Maija J.Lahtinen Anu Suoranta Bishwa Ghimire Pirkko Mattila Johanna Uusimaa Markku Varjosalo Jani Katisko Reetta Hinttala PEDEGO Research Unit University of OuluOuluFinland Medical Research Center Oulu University HospitalUniversity of OuluOuluFinland Biocenter Oulu University of OuluOuluFinland Institute of Biotechnology HiLIFE Helsinki Institute of Life ScienceUniversity of HelsinkiHelsinkiFinland Drug Research Program Faculty of PharmacyUniversity of HelsinkiHelsinkiFin-land Neurosurgery Research Unit of Clinical NeuroscienceOulu University HospitalUniversity of OuluOuluFinland Oulu Research Group of Advanced Surgical Technologies and Physics(ORGASTP) Research Unit of Clinical NeuroscienceOulu University HospitalUniversity of OuluOuluFinland Institute for Molecular Medicine Finland(FIMM) HiLIFE Helsinki Institute of Life ScienceUniversity of HelsinkiHelsinkiFinland Clinic for Children and Adolescents Division of Pediatric NeurologyOulu University HospitalOuluFinland

Background:Transcriptomic and proteomic profiling of human brain tissue is hindered by the availability of fresh samples from living patients.Postmortem samples usually represent the advanced disease stage of the patient.Fur-thermore,the postmortem interval can affect the transcriptomic and proteomic profiles.Therefore,fresh brain tissue samples from living patients represent a valuable resource of metabolically intact tissue.Implantation of deep brain stimulation(DBS)electrodes into the human brain is a neurosurgical treatment for,e.g.,movement disorders.Here,we describe an improved approach to collecting brain tissues from surgical instruments used in implantation of DBS device for transcriptomics and proteomics analyses.Methods:Samples were extracted from guide tubes and recording electrodes used in routine DBS implantation procedure to treat patients with Parkinson’s disease,genetic dystonia and tremor.RNA sequencing was performed in tissues extracted from the recording microelectrodes and liquid chromatography-mass spectrometry(LC-MS)per-formed in tissues from guide tubes.To assess the performance of the current approach,the obtained datasets were compared with previously published datasets representing brain tissues.Results:Altogether,32,034 RNA transcripts representing the unique Ensembl gene identifiers were detected from eight samples representing both hemispheres of four patients.By using LC-MS,we identified 734 unique proteins from 31 samples collected from 14 patients.The datasets are available in the BioStudies database(accession number S-BSST667).Our results indicate that surgical instruments used in DBS installation retain brain material sufficient for protein and gene expression studies.Comparison with previously published datasets obtained with similar approach proved the robustness and reproducibility of the protocol.Conclusions:The instruments used during routine DBS surgery are a useful source for obtaining fresh brain tis-sues from living patients.This approach overcomes the issues that arise

关键词： Deep brain stimulation Movement disorders Brain Proteomics Transcriptomics RNA sequencing LC-MS Personalized medicine

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基于^(1)H-NMR结合LC-MS技术分析熊胆粉及常见胆类中药中的胆酸类成分

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中草药 2022年第17期53卷 5283-5292页

作者：曹妍张佩杰李婷许霞常安琪李军刘地发屠鹏飞宋月林北京中医药大学中药学院中药现代研究中心北京100029 北京中医药大学中药品质评价北京市重点实验室北京100029 江西青峰药业有限公司创新天然药物与中药注射剂国家重点实验室江西赣州341000

目的建立基于核磁共振氢谱技术(^(1)H-NMR)结合液质联用技术(LC-MS)的分析方法,阐明熊胆粉中胆酸类成分组成,找出熊胆粉与其他常见胆类中药中的差异性成分。方法利用^(1)H-NMR非靶标分析熊胆粉及其他常见胆类中药的化学成分组,通过信... 详细信息

目的建立基于核磁共振氢谱技术(^(1)H-NMR)结合液质联用技术(LC-MS)的分析方法,阐明熊胆粉中胆酸类成分组成,找出熊胆粉与其他常见胆类中药中的差异性成分。方法利用^(1)H-NMR非靶标分析熊胆粉及其他常见胆类中药的化学成分组,通过信号归属表征熊胆粉中胆酸类成分组成,以^(1)H-NMR图谱分段积分后峰面积为变量进行多元统计分析。利用LC-MS对熊胆粉及其他常见胆类中药中10个胆酸类成分(胆酸、熊去氧胆酸、猪去氧胆酸、鹅去氧胆酸、牛磺熊去氧胆酸、牛磺鹅去氧胆酸、甘氨熊去氧胆酸、甘氨猪去氧胆酸、牛磺胆酸、甘氨胆酸)进行含量测定,并以含量为变量进行多元统计分析。结果基于^(1)H-NMR图谱及多元统计分析发现胆酸类成分为胆类中药的主要成分,也是熊胆粉与其他常见胆类中药的主要差异性成分。结合文献及对照品比对,从熊胆粉中初步鉴定出7个胆酸类成分。进一步结合LC-MS含量测定结果和多元统计分析验证熊胆粉与其他常见胆类中药的主要差异性胆酸类成分为熊去氧胆酸、鹅去氧胆酸、牛磺熊去氧胆酸和牛磺鹅去氧胆酸。结论熊胆粉与其他常见胆类中药的胆酸类成分存在显著差异。与其他胆类中药相比,熊胆粉中熊去氧胆酸、鹅去氧胆酸及其牛磺型结合胆酸含量较高。^(1)H-NMR技术具有对化学成分组全面、快速分析的优势,可与LC-MS检测相互补充,共同应用于中药复杂体系成分分析和代谢组学的深入研究。

关键词： ^(1)H-NMR LC-MS 熊胆粉常见胆类中药胆酸类成分多元统计分析熊去氧胆酸鹅去氧胆酸牛磺熊去氧胆酸牛磺鹅去氧胆酸

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藏族药二十五味松石丸治疗慢性胆汁淤积小鼠的血清代谢组学研究

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中国中药杂志 2022年第8期47卷 2056-2063页

作者：李阿溶王存萍丁翼简程芳张乐顾健谭睿西南民族大学药学院四川成都610041 西南交通大学生命科学与工程学院四川成都610031

采用喂养含3,5-二乙氧基羰基-1,4-二氢-2,4,6-三甲基吡啶(3,5-diethoxy carbonyl-1,4-dihydro-2,4,6-trimethylpyridine,DDC)的饲料建立慢性胆汁淤积小鼠模型,基于液相色谱-质谱(LC-MS)检测技术的代谢组学分析方法,探究二十五味松石丸(Ershiwuwei Songshi Pills,ESP)对慢性胆汁淤积小鼠的内源性代谢物的影响。结果发现ESP能够有效改善模型组小鼠的病理性损伤以及降低血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、总胆汁酸(TBA)水平。代谢组学筛查得到模型组与空白组、模型组与ESP组13个共有的差异代谢物,分别是尿酸(uric acid)、乙醇醛(glycolaldehyde)、犬尿氨酸(kynurenine)、黄素腺嘌呤二核苷酸(flavin adenine dinucleotide)、L-3-苯乳酸(L-3-phenyllactic acid)、I-尿胆素(I-urobilin)、白三烯D4(LTD4)、牛磺胆酸(taurocholic acid)、trioxilin A3、1D-肌醇1,4-二磷酸(D-inositol-1,4-diphosphate)、PC[16:0/20:2(11Z,14Z)]、PC[14:0/22:2(13Z,16Z)]、PC[20:4(5Z,8Z,11Z,14Z)/20:4(5Z,8Z,11Z,14Z)]。ESP干预后,13个差异代谢物的含量均有明显回调,通路分析显示,ESP主要通过影响花生四烯酸代谢、甘油磷脂代谢、色氨酸代谢、初级胆汁酸的生物合成等途径来达到治疗效果。该研究从代谢物角度阐释了二十五味松石丸抗慢性胆汁淤积的作用机制。

关键词：二十五味松石丸胆汁淤积 LC-MS 代谢组学

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