运用液相色谱联合离子阱-飞行时间质谱法进行海洛因成瘾人员代谢组学研究

利用代谢组学的研究方法,对海洛因成瘾人员尿液及血清中可能的相关标志物进行筛选。运用液相色谱联合离子阱-飞行时间质谱(LC/MS-IT-TOF)联用技术对海洛因检测尿检条阴性的16名海洛因滥用成瘾者与16名正常人的血清和尿液进行研究。多变量统计分析结果表明,海洛因成瘾者和正常人聚类明显,血清和尿液中分别发现12种可能的潜在标志物。成瘾人员和正常人员在血清及尿液代谢水平上具有明显差异,差异代谢物的发现有助于为发现海洛因成瘾判定的潜在标志物提供依据。     

亲水作用色谱/质谱联用方法用于膀胱癌患者血清代谢组学研究

膀胱癌是泌尿系统最常见的恶性肿瘤之一,具有高发病率、高复发率和高进展率的特点.本研究应用69个极性代谢物标样选择合适的分离系统,建立了两性离子亲水作用色谱/质谱联用的代谢组学分析方法.本方法线性范围较宽,检出限低于ng/mL数量级.将本方法用于血清代谢组学分析,85%以上代谢物峰面积的RSD<30%.对64例膀胱癌患者和32例正常人的血清进行代谢组学研究,发现溶血磷脂酰胆碱、游离脂肪酸、氨基酸、胆汁酸、有机酸、核苷等在患病组和正常组中存在显著差异.经筛选和验证,甘磷酸胆碱、胱氨酸、十二碳烯酸、二十碳烯酸和鹅去氧胆酸5种代谢物可以作为区分膀胱癌和正常人的潜在标志物.本研究结果表明,基于亲水作用色谱/质谱联用的代谢组学方法是发现癌症诊断潜在生物标志物的有效手段.     

超高效液相色谱-四极杆-静电场轨道阱质谱联用法用于宫颈癌患者尿液代谢组学研究

利用超高效液相色谱-四极杆-静电场轨道阱质谱联用(UHPLC-Q-Orbitrap MS)方法对宫颈癌(Cervical cancer,CC)患者和健康人(Healthy control,HC)的尿液进行分析,研究宫颈癌患者尿液中的潜在标志物,为其发病机制和诊断提供科学依据。筛选11例宫颈癌患者(Age(45.7±5.6)years)及11例健康人(Age(45.9±3.2)years)尿液样本,采用液相色谱-质谱联用技术对尿液进行测定,通过主成分分析(PCA)和偏最小二乘法-判别分析(OPLS-DA)处理数据,结果表明,两组人群代谢轮廓有显著差别,发现并鉴定了12种潜在的生物标志物,提示特定的肿瘤代谢途径中潜在的代谢标志物可能在宫颈癌发生发展中发挥重要作用。     

短暂性脑缺血的尿液代谢标志物筛选

以筛选短暂性脑缺血(TIA)的早期体内生物标志物为目的,基于高分离度液相色谱-四极杆飞行时间质谱联用技术对双侧颈总动脉闭塞引起的TIA大鼠模型的尿液进行代谢组学研究。筛选并鉴定L-苹果酸、D-葡萄糖醛酸-6,3-内酯、L-苏氨酸、L-缬氨酸、L-古洛糖酸-1,4-内酯、抗坏血酸、磷酸烯醇丙酮酸7种TIA损伤重要的尿液代谢标志物;主要参与机体的抗坏血酸代谢、丙酮酸代谢与支链氨基酸代谢,调节体内的氧化应激、炎症反应与神经保护。本研究运用了高分离度色谱质谱联用技术筛选TIA代谢标记物,为脑卒中的早期诊断与治疗提供了科学依据。     

基于液相色谱/电喷雾飞行时间质谱技术的结石症尿液代谢组学探讨

探索了一种简便的尿液样品前处理流程,通过反相液相色谱(RPLC)与亲水相互作用色谱(HILIC)相结合的高效液相色谱/电喷雾飞行时间质谱的联用技术(HPLC/ESI-TOFMS),建立了泌尿系结石症的尿液代谢组学模型,研究结石症患者尿液样品中的代谢物浓度水平与健康人对照组间的差异情况。采用主成分分析(PCA)及偏最小二乘法-判别分析(PLS-DA)等多变量数据分析方法,寻找结石症患者组和对照组之间的差异性代谢谱。模型结果显示:结石症患者尿液代谢物水平与健康对照组具有明显差异,寻找其中潜在的差异性标志物群组,有望将该模型用于泌尿系结石症的快速预测和诊断。     

基于拟靶向液相色谱-质谱联用的胃癌患者血清代谢组分析

胃癌是一种高发的恶性肿瘤,是癌症相关死亡的第二大病因。早期筛查是提高患者生存率的有效手段,但目前临床上尚缺乏实现胃癌无创筛检的可靠标志物。本研究采用了基于液相色谱-质谱联用的拟靶向代谢组学方法分析了20例胃癌患者及40例正常人血清代谢组,以期发现新的潜在代谢标志物。代谢组数据的主成分分析和偏最小二乘法数据分析结果显示,胃癌患者与健康人群的血清代谢组存在明显的差异,结合非参数检验进一步筛选并定性出57个差异代谢物。其中二氢胆固醇经验证组样本验证,具有成为胃癌代谢标志物的潜力。本研究在发现胃癌的潜在代谢标志物的同时,也为胃癌患者代谢分型提供了重要的科学依据。     

基于液相色谱-质谱技术的代谢组学分析方法新进展

液相色谱-质谱联用技术是代谢组学研究领域的主要技术平台之一,近年来基于液相色谱-质谱联用技术的代谢组学分析方法获得了巨大发展。本文结合本研究组在代谢组学方面的研究成果,综述了近年来液相色谱-质谱联用技术在代谢组学分析方法方面的新进展,并对其发展前景进行了展望。综述引用文献41篇。     

一种合成亚洲玉米螟性外激素的新方法

本文报道了一种合成亚洲玉米螟(Ostrinia furnacalis Guenee)性外信息素新方法:即将10-十一碳烯-1-醇乙酸酯经硼氢化反应得到12-十四碳炔-1-醇乙酸酯,再进行定向还原反应.这样就将原来的五步合成反应缩短到两步.最后分别得到高纯度的(E)和(Z)-12-十四碳烯-1-醇乙酸酯.产品中双键的位置借毛细管气相色谱仪和气相色谱-质谱联用仅得到了确定.反应中间体12-十四碳炔-1-醇也可从2-十三碳炔-1-醇经过“炔键移位反应”(Zipper reaction)、甲基化和乙酰化反应而得到。     

基于UPLC-oaTOF-MS的糖尿病及糖尿病肾病的代谢组学研究

以糖尿病患者、糖尿病肾病患者和正常人的血清为研究对象, 采用超高效液相色谱-飞行时间质谱建立其代谢指纹图谱, 并结合主成分分析进行模式识别, 实现患者和正常人的区分, 并试图发现潜在的标志物.     

基于快速高分辨液相色谱串联质谱技术的代谢组学尿液分析方法的建立

采用快速高分辨液相色谱(RRLC)分离系统与QTRAP型及QTOF型MS/MS仪联用技术,通过考察尿液样本前处理方法,优化液相色谱条件和质谱检测参数,建立了用于尿液中代谢物分析的RRLC-MS方法.采用本方法对尿液浓度下的20种代表性代谢物进行了检测,考察了方法的灵敏度和精密度,证明本方法适用于尿液代谢组学的研究.对穿...     

慢性心功能衰竭患者尿液的代谢组学研究

采用基于液相色谱-质谱联用的方法对慢性心力衰竭(Chronic heart failure, CHF)患者和正常对照(Control)人群的尿液进行分析, 筛选慢性心力衰竭患者尿液中的差异代谢物, 研究其发病机制, 并为临床治疗提供科学依据.选择15个慢性心力衰竭患者(年龄(62.27±3.14)岁)及15个正常人(年龄(65.41±4.63)岁), 采用高分辨度快速液相色谱-四极杆-飞行时间串联质谱(RRLC-QTOF/MS)技术对尿液代谢物进行分析, 采用主成分分析(PCA)对两组代谢物进行分类, 并筛选潜在生物标记物;运用偏最小二乘判别分析法(PLS-DA)建模, 考察生物标记物对疾病筛选的预测能力.研究结果表明, CHF组和Control组尿液代谢物谱能得到很好的区分, 发现并鉴定了2种潜在生物标记物尿苷及丙氨酰色氨酸, 提示嘧啶代谢和色氨酸代谢可能在心力衰竭发生发展中有重要作用.     

小儿肺炎患者尿液的代谢组学初步研究

利用液相色谱-质谱联用法对小儿肺炎( Childhood pneumonia, CP)患者和健康儿童( Healthy control)的尿液进行分析,发现小儿肺炎患者尿液中的潜在标记物,为其发病机制及早期筛查提供科学依据。筛选10例小儿肺炎患者(age 47.72±2.35 months)及10例健康儿童(age 46.65±1.97 months)尿液样本,采用快速高分辨液相色谱四极杆-飞行时间质谱联用( RRLC-Q TOF/MS)技术对其尿液代谢物进行分析,通过主成分分析方法( PCA)对两组代谢物进行分类,并发现潜在生物标记物。 RRLC-Q TOF/MS检测表明,CP组和Healthy Control组尿液代谢物图谱能得到很好的区分,并鉴定了5种生物标记物,提示嘌呤代谢、氨基酸代谢可能在小儿肺炎发生发展中有重要作用。     

Urinary Metabonomics Study of Heart Failure Patients with HILIC and RPLC Separation Coupled to TOF–MS

In this study, urinary metabolic profiles of patients with heart failure (HF) and healthy individuals were analyzed by LC-TOF–MS. Both reversed-phase chromatography and hydrophilic interaction chromatography were used to separate the endogenous metabolites in urine. Partial least-squares to latent structure-discriminant analysis was used for discriminating HF patients from healthy persons and the selection of potential biomarkers. The results suggested that the combination of LC–MS and multivariate statistical analysis could be used for HF diagnosis. The MS/MS experiments were carried out to identify the potential biomarkers which are important for the contribution to the discrimination. As a result, 12 potential biomarkers for HF were identified and the related metabolic pathways were studied.     

Urinary metabolomic study of systemic lupus erythematosus based on gas chromatography/mass spectrometry

Systemic lupus erythematosus (SLE) is an autoimmune disease with heterogeneous organ and system manifestations. In this study, urinary metabolic alterations related to SLE were investigated by performing gas chromatography/mass spectrometry (GC/MS) based metabolomics and multivariate statistical analysis. Patients with SLE and healthy controls could be clearly differentiated in view of the metabolic abnormity in urine. Among 70 identified endogenous metabolites, 23 metabolites were dramatically increased in SLE patients, which involved in several key metabolic pathways including energy metabolism, nucleotide metabolism, oxidative stress and gut‐microbiome‐derived metabolism. This noninvasive and GC/MS‐based metabolomic technique is a promising and potent strategy for identifying novel biomarkers and understanding pathogenesis of SLE. Copyright © 2016 John Wiley & Sons, Ltd.     

Metabolomics approach based on utra-performance liquid chromatography coupled to mass spectrometry with chemometrics methods for high-throughput analysis of metabolite biomarkers to explore the abnormal metabolic pathways associated with myocardial dysfunction

Ultra-performance liquid chromatography/mass spectrometry-based metabolomics can been used for discovery of metabolite biomarkers to explore the metabolic pathway of diseases. Identification of metabolic pathways is key to understanding the pathogenesis and mechanism of disease. Myocardial dysfunction induced by sepsis (SMD) is a severe complication of septic shock and represents major causes of death in intensive care units; however its pathological mechanism is still not clear. In this study, ultrahigh-pressure liquid chromatography with mass spectrometry-based metabolomics with chemometrics anaylsis and multivariate pattern recognition analysis were used to detect urinary metabolic profile changes in a lipopolysaccharide-induced SMD mouse model. Multivariate statistical analysis including principal component analysis and orthogonapartial least squares discriminant analysis for the discrimination of SMD was conducted to identify potential biomarkers. A total of 19 differential metabolites were discovered by high-resolution mass spectrometry-based urinary metabolomics strategy. The altered biochemical pathways based on these metabolites showed that tyrosine metabolism, phenylalanine metabolism, ubiquinone biosynthesis and vitamin B6 metabolism were closely connected to the pathological processes of SMD. Consequently, integrated chemometric analyses of these metabolic pathways are necessary to extract information for the discovery of novel insights into the pathogenesis of disease.     

Plasma metabolomic profile and potential biomarkers for missed abortion

A missed abortion (MA) is an in utero death of the embryo or fetus before the 20th week of gestation with retained products of conception, and this condition is currently common in China. In order to discover novel biomarkers for MA, ultrahigh performance liquid chromatography was applied to study plasma metabolite profiles for 33 patients with MA and 29 control subjects. Thirty‐seven differential plasma metabolites were found to discriminate between the two groups in the initial cohort (15 subjects with MA and 15 healthy controls). The feasibility of using these potential biomarkers to predict MA was further evaluated in the validation cohort (18 subjects with MA and 14 healthy controls) and 15 had an area under the receiver operating characteristic curve of >0.80, making them satisfactory. Tryptophan metabolism and sphingolipid metabolism were identified as important potential target pathways for MA using metabolic pathway impact analysis. Furthermore, three of the 15 satisfactory metabolites (glyceric acid, indole and sphingosine) were combined to establish a predictive model with 100% sensitivity and 100% specificity in the validation cohort. Taken together, these results suggest that MA results in significant disturbance of metabolism and those various novel biomarkers have satisfactory diagnostic and predictive power for MA.     

Lysophosphatidylcholine Biomarkers of Lung Cancer Detected by Ultra-performance Liquid Chromatography Coupled with Quadrupole Time-of-flight Mass Spectrometry

Centrifugal ultrafiltration after methanol extraction of whole plasma was used as an optimal condition for the preparation of blood plasma before metabonomic studies. The plasma samples from 102 lung cancer patients and 34 healthy volunteers were prepared with this approach. With ultra-performance liquid chromatography(UPLC) coupled with quadrupole time-of-flight mass spectrometry(Q-TOF MS) analysis, the samples were investigated in order to find potential disease biomarkers. After data acquisition, orthogonal signal correction partial least squares models were built to differentiate the healthy volunteers from lung cancer patients and to identify metabolites that showed significantly different expression between the two groups. Several metabolite ions were identified as potential biomarkers according to the variable importance in the project(VIP) value in both ion modes. Five lysophosphatidylcholines were further identified as specifically lysoPC 16:0, isomer of lysoPC 16:0, lysoPC 18:0, lysoPC 18:1 and lysoPC 18:2. These results suggest that UPLC coupled with Q-TOF MS is an effective technique for the analysis of plasma metabolites in metabonomic studies.     

Metabonomics analysis of semen euphorbiae and semen Euphorbiae Pulveratum using UPLC–Q-TOF/MS

Semen Euphorbiae (SE), the dry and mature seed of Euphorbia lathyris L., a common traditional Chinese medicine, has significant pharmacological activity. However, its toxicity limits its clinical application, and less toxic Semen Euphorbiae Pulveratum (SEP) is often used clinically. To explore the possible mechanism of SE frost-making and attenuation, this study used ultrahigh-performance liquid chromatography coupled with quadrupole time of flight mass spectrometry to perform a comprehensive metabolomics analysis of serum and urine samples from rats treated with SE and SEP, and performed histopathological evaluation of liver, kidney and colon tissues. Meanwhile, the different metabolites were visualized through multivariate statistical analysis and the HMDB and KEGG databases were used to distinguish the differential metabolites of SE and SEP to reveal related metabolic pathways and their significance. In total, 32 potential biomarkers, 14 in serum and 18 in urine, were identified. The metabolic pathway analysis revealed that arachidonic acid metabolism, sphingolipid metabolism, tyrosine and tryptophan biosynthesis, the tricarboxylic acid cycle and seven other metabolic pathways were significantly altered. Importantly, compared with SE, SEP reduced the metabolic disorder related to endogenous components. The mechanism may be related to the regulation of lipid metabolism, intestinal flora metabolites, amino acid metabolism and energy metabolism. This study provided new insights into the possible mechanism of SE freezing and attenuation.     

基于快速高分辨液相色谱-质谱的中波紫外线辐射诱导大鼠急性光损伤的代谢组学研究

利用快速高分辨液相色谱-四极杆-飞行时间质谱( RRLC-Q-TOF-MS)联用技术结合多元统计分析方法,考察在中波紫外线( Ultraviolet B, UVB)辐射前后,大鼠尿液中内源性代谢物谱的变化,研究UVB辐射导致急性光损伤的生理机制。急性光损伤大鼠模型由窄谱中波紫外线光源(TL-01,峰值312 nm)照射,采用离心沉降后四倍稀释法处理尿液样本, Supelco Ascentis? Express C18色谱柱,水(含0.1％甲酸)与乙腈为流动相梯度洗脱,液相色谱-串联质谱分析测定。利用主成分分析( PCA)法、聚类分析( CA)法等对辐射前后的大鼠尿液样本进行代谢轮廓分析,寻找对分组贡献大的差异代谢物及通路,并阐明其作用机制;运用偏最小二乘判别分析( PLS-DA)法建立预测模型,考察此模型在UVB致光损伤模型诊断上的预测能力。多元统计分析结果显示,空白对照组与UVB模型组能够获得很好地区分,通过将差异代谢物与数据库、串联质谱数据及标准品比对,发现并鉴定出11种潜在生物标记物,表明UVB辐射可影响正常大鼠的鞘脂类代谢、核酸代谢、亚油酸代谢、氨基酸代谢等通路,这些差异代谢物对UVB辐射致光损伤类疾病的诊断具有较好的预判能力。     

Serum metabolomics of laryngeal cancer based on liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry

The discovery of new laryngeal cancer‐related metabolite biomarkers could help to facilitate early diagnosis. A serum metabolomics study from laryngeal cancer patients and healthy individuals was conducted using liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry. Univariate and multivariate statistics were used to discriminate laryngeal cancer patients and healthy individuals. 1‐Palmitoyl‐sn‐glycero‐3‐phosphocholine (LysoPC 16:0), 1‐o‐hexadecyl‐2‐acetyl‐sn‐glycero‐3‐phosphocholine (PAF) and 1,2‐dipalmitoyl‐sn‐glycero‐3‐phosphocholine were found to be significantly different between the laryngeal cancer group and the healthy group. They are mainly involved in phospholipids catabolism, linoleic acid metabolism, α‐linoleic acid metabolism and arachidonic acid metabolism. The area under the curve of the biomarker combined by two metabolites (LysoPC 16:0 and PAF) was 0.935, the sensitivity was 0.962 and the specificity was 0.825. LysoPC 16:0 and PAF may show diagnostic potential for laryngeal carcinoma.