基于呼出气挥发性有机物检测的肺癌筛查技术研究进展

肺癌已成为当前全球发病率和死亡率最高的癌症.早期肺癌的治愈率高,"早诊早治"已成为共识.呼出气检测具有非侵入性、取样方便且对人体友好的优势,通过对呼出气中特征代谢物的检测,寻找早期肺癌的疾病标志物,有望作为一种简单有效的早期肺癌筛查手段而备受关注.本文从临床常用肺癌筛查方法、呼出气组分及肺癌标志性挥发性有机物(VOCs...     

人体呼出气分析的技术进展及其在非侵入式医学诊断方面的临床应用前景

人体呼出气被视为血液顶空气体,通过肺泡交换排出体外,可在一定程度上反映人体的内源代谢情况。近些年,随着医学诊断无损化的不断发展,呼出气分析由于其无创性,无痛性,便捷性,具有临床疾病早期诊断与大规模筛查的潜力,受到越来越多的关注。研究表明,呼出气中包含的挥发性有机化合物和人体的疾病代谢状态密切相关,如丙酮与糖尿病,醛类物质与乳腺癌,烷烃类与氧化应激水平等。现代分析方法(如气相色谱、质谱、光谱、传感器等)对呼出气中上千种痕量组分的定性定量分析,使得呼出气分析成为可能。此文总结了呼出气分析的技术发展:样品采集手段,预富集技术和定性定量分析技术。评价了呼出气生物标识物在糖尿病,乳腺癌,以及肺癌等疾病中的临床诊断前景。最后讨论了呼出气分析的技术在国内的发展现状及其面临的主要问题。     

二次纳流电喷雾电离耦合超高分辨质谱检测人体呼出气中相对高分子量化合物

二次电喷雾电离源耦合超高分辨质谱(SESI-UHRMS)有望检出人体呼出气中分子量大于300的相对高分子量化合物,这些化合物的发现将有助于更准确地理解呼出气中挥发性有机化合物(VOCs)的来源、产生机制以及SESI源电离机理,更好地实现SESI-UHRMS的转化应用.本研究自组装nanoSESI源(尚无商业产品)耦合四极杆-静电场轨道阱质谱(最高质量分辨率1.2×10~5),考察了该装置对健康人体呼出气中分子量为300~500化合物的检出情况.结果表明,所搭建的nanoSESI-UHRMS装置检测人体呼出气的重现性好、灵敏度高,可检出数十种分子量为300~500的化合物.根据这些化合物在单次呼气过程中信号强度随时间变化的趋势,推断其来源分别为内源性和外源性;各化合物的元素组成主要包括C,H,N和O,环-双键当量(RDB)的均值为(4.5±3.1),表明检出的化合物可能为醛酮或不饱和脂肪酸,容易在SESI源中被电离.本研究初步验证了自组建nanoSESI-UHRMS检测人体呼出气中相对高分子量化合物的可行性,为后续进一步开展方法应用奠定了基础.     

白云鄂博矿工人呼出气中(气土)活度的测定结果及卫生学评价

本文介绍用负高压静电收集和测量人体呼出气中氢子体置装,测定了白云鄂博矿按比例分层随机抽样的130名接触粉尘工人呼出气中的氢活度,并据此推算出肺内天然钍的沉积量。对其卫生意义也作出了评论。     

燃料电池型酒精检测仪

燃料电池型酒精检测仪是根据电池工作原理检查人体呼出气中的酒精含量,再换算成人体血液中的酒精含量,为检测驾驶员酒后开车、人体酒精中毒提供了一种简便、定量的手段。     

冷凝收集-离子色谱法测定呼出气中的有机酸和阴离子

建立了一种非侵入式冷凝收集-离子色谱方法测定人体呼出气中乳酸、甲酸、乙酸、丙酮酸、Cl^-、NO₂^-、NO₃^-、SO₄^2-。搭建自制呼出气冷凝装置,该装置包括吹气口、与吹气口相连的单向阀和流量计、置于半导体冷凝装置中的冷阱以及一次性冷凝收集管。通过呼出气冷凝装置对人体呼出气进行收集,利用离子色谱对冷凝液(EBC)中有机酸和阴离子的含量进行检测。优化采集冷阱温度和采集流量,得到冷阱最佳冷凝温度为-15℃,呼气流量为15 L/min。采用1.5 mmol/L碳酸钠和3 mmol/L碳酸氢钠混合溶液作为流动相,泵流速为0.8 mL/min,分析柱为IC-SA3 (250 mm×4.0 mm),柱温为45℃。8种有机酸和阴离子的线性范围均为0.1～10.0 mg/L,相关系数均≥0.999 3。在进样量为100μL时,方法的检出限为0.001 7～0.015 0 mg/L(S/N=3),定量限为0.005 7～0.050 0 mg/L(S/N=...     

气敏新材料MXenes在呼出气体传感器中的应用

电子鼻结合人工智能对呼出气进行检测、分析和识别已成为非侵入性医疗检测领域的研究热点.然而,目前已报道的气体传感材料尚不能同时满足高灵敏度、高选择性和稳定的室温检测,阻碍了气体传感器在医疗健康领域的应用及发展,寻找合适的传感材料具有重要的意义和挑战.新型二维层状纳米材料MXenes具有种类多、比表面积大、导电性能强、表面...     

NEWS

人体新陈代谢的部分产物由血液运送至肺部,在肺泡通过气体交换出现在呼出气体中。呼出气体反映了机体的代谢和病理状况,一些物质甚至可能成为某种疾病的生物标记物。如：烷烃类是氧化应激的标志物,丙酮是糖尿病的生物标志物等。近年来,在气道慢性炎症,肺癌,肺结核,肠病,肝硬化,乳腺癌,胃管-食管癌等疾病中利用呼出气体检测来寻找特异性标志物的研究越来越多。国际上为此成立了专门的呼出气研究协会( International Association of Breath Research, IABR),并出版了呼出气体检测的专业性杂志Journal of breath research。     

基于单颗粒气溶胶质谱的人体呼出颗粒物粒径分布与化学成分的分析方法研究

人体呼出气中内源性颗粒物和外源性颗粒物的粒径和化学成分信息可为肺部疾病诊断、环境暴露评价等研究提供参考。该文初步考察了单颗粒气溶胶质谱(Single particle aerosol mass spectrometry,SPAMS)同时获取人体呼出气中颗粒物(Exhaled breath particles,EBPs)粒径分布和化学成分的可行性。结果表明,健康成人的EBPs数浓度为227~1 043个/L,获取具有统计意义的粒径分布所需的EBPs检出限为2 500个颗粒物,粒径范围为200~1 000 nm,峰值出现在460 nm。与环境空气颗粒物的粒径分布相比,EBPs更多分布在200~300 nm和440~660 nm,从化学成分来看,这两段粒径范围内的EBPs含有更多的碳元素,不易在体内发生吸湿增长,提高了被呼出的概率。EBPs的化学成分可能反映内源性颗粒物和外源性颗粒物组成,如HSO_4~-、PO_3~-、CN~-、CNO~-和C_xH_yO_z~+(x=1~3,y=1~7,z=1~3)可能与内源性颗粒物中的蛋白含量、磷酸酯酰甘油等成分有关,碳簇峰C_3~-、C_4~-、C~+、C_3~+、C_3H~+和C_4~+推测与外源性颗粒物中的碳元素有关。     

基于呼气中特异性VOCs筛查的肺癌早期诊断与模型评估

采用气相色谱-质谱联用仪（GC-MS）检测了180例受试者的呼出气样品,包括79名肺癌患者和101名健康志愿者。每个受试者采集3个平行样品,以及1个室内空气样品。对所有呼气样品中检测的92种挥发性有机物（VOCs）进行定量分析。结合Mann-Whitney检验和正交偏最小二乘判别分析（OPLS-DA）模型筛选出10种肺癌患者呼气特异性VOCs,包括苯甲醛、顺式-2-丁烯、2-丁酮、萘、乙酸乙烯酯、乙烯、2,2,4-三甲基戊烷、3-甲基戊烷、己醛和2-甲基戊烷。利用统计学方法研究其在不同人群中的代谢差异和可能相关的代谢机制,通过建立机器学习模型验证候选标志物对疾病的诊断性能,结果显示,随机森林模型诊断的准确度、精准率、灵敏度和特异性分别为96.25%、96.21%、95.76%、96.67%,马修斯相关系数（MCC）为0.93,曲线下面积为0.96。上述10种化合物可作为肺癌患者的潜在呼气VOCs标志物,为肺癌的早期诊断提供了丰富的基础数据。     

Comprehensive Two-Dimensional Gas Chromatography–Mass Spectrometry Analysis of Exhaled Breath Compounds after Whole Grain Diets

Exhaled breath is a potential noninvasive matrix to give new information about metabolic effects of diets. In this pilot study, non-targeted analysis of exhaled breath volatile organic compounds (VOCs) was made by comprehensive two-dimensional gas chromatography–mass spectrometry (GCxGC-MS) to explore compounds relating to whole grain (WG) diets. Nine healthy subjects participated in the dietary intervention with parallel crossover design, consisting of two high-fiber diets containing whole grain rye bread (WGR) or whole grain wheat bread (WGW) and 1-week control diets with refined wheat bread (WW) before both diet periods. Large interindividual differences were detected in the VOC composition. About 260 VOCs were detected from exhaled breath samples, in which 40 of the compounds were present in more than half of the samples. Various derivatives of benzoic acid and phenolic compounds, as well as some furanones existed in exhaled breath samples only after the WG diets, making them interesting compounds to study further.     

Human exhaled air analytics: biomarkers of diseases

Over the last few years, breath analysis for the routine monitoring of metabolic disorders has attracted a considerable amount of scientific interest, especially since breath sampling is a non-invasive technique, totally painless and agreeable to patients. The investigation of human breath samples with various analytical methods has shown a correlation between the concentration patterns of volatile organic compounds (VOCs) and the occurrence of certain diseases. It has been demonstrated that modern analytical instruments allow the determination of many compounds found in human breath both in normal and anomalous concentrations. The composition of exhaled breath in patients with, for example, lung cancer, inflammatory lung disease, hepatic or renal dysfunction and diabetes contains valuable information. Furthermore, the detection and quantification of oxidative stress, and its monitoring during surgery based on composition of exhaled breath, have made considerable progress. This paper gives an overview of the analytical techniques used for sample collection, preconcentration and analysis of human breath composition. The diagnostic potential of different disease-marking substances in human breath for a selection of diseases and the clinical applications of breath analysis are discussed.     

纳米材料在用于癌症诊断的呼出气挥发性有机物检测中的应用

细胞新陈代谢的变化会导致挥发性有机化合物（VOCs）类型及含量发生变化,因此可通过分析某些标志性VOCs简立起多种疾病早期诊断的模型. 人体呼出物中特征VOCs的检测作为一种非侵入性、无损的检测手段,近些年在疾病检测领域已成为世界范围内的研究热点. 其中,纳米材料可用于增强传感器性能,并使传感器便携式小型化,推进检测传感器进入临床. 在这篇综述中,作者将种类繁多的传感器中用到的纳米材料归纳总结为金属、金属氧化物、碳基、复合物和MOFs基纳米材料等几类,并讨论了不同类纳米材料在VOCs检测中的优劣势. 本文所建立起的分析方法及讨论有助于进一步了解检测技术的优越性与局限性. 最后,作者对利用VOCs的检测实现癌症早期筛选的研究及发展提出了个人观点.     

Ultrafast gas chromatography coupled to electronic nose to identify volatile biomarkers in exhaled breath from chronic obstructive pulmonary disease patients: A pilot study

An analytical method to identify volatile organic compounds (VOCs) in the exhaled breath from patients with a diagnosis of chronic obstructive pulmonary disease (COPD) using a ultrafast gas chromatography system equipped with an electronic nose detector (FGC eNose) has been developed. A prospective study was performed in 23 COPD patients and 33 healthy volunteers; exhalation breathing tests were performed with Tedlar bags. Each sample was analyzed by FCG eNose and the identification of VOCs was based on the Kovats index. Raw data were reduced by principal component analysis (PCA) and canonical discriminant analysis [canonical analysis of principal coordinates (CAP)]. The FCG eNose technology was able to identify 17 VOCs that distinguish COPD patients from healthy volunteers. At all stages of PCA and CAP the discrimination between groups was obvious. Chemical prints were correctly classified up to 82.2%, and were matched with 78.9% of the VOCs detected in the exhaled breath samples. Receiver operating characteristic curve analysis indicated the sensitivity and specificity to be 96% and 91%, respectively. This pilot study demonstrates that FGC eNose is a useful tool to identify VOCs as biomarkers in exhaled breath from COPD patients. Further studies should be performed to enhance the clinical relevance of this quick and ease methodology for COPD diagnosis.     

Volatile Organic Compounds Analysis in Breath Air in Healthy Volunteers and Patients Suffering Epidermoid Laryngeal Carcinomas

Exhaled breath contains thousands of gaseous volatile organic compounds (VOCs) that may be used as non-invasive markers of head and neck epidermoid cancer. We hypothesized that solid phase micro-extraction coupled to gas chromatography–mass spectrometry can discriminate patients with epidermoid head and neck cancer from healthy controls by analyzing the gaseous volatile organic compounds, VOC-profile, in exhaled breath, thus identifying some non-invasive biomarkers to be used in early detection. Twenty healthy subjects participated in a cross-sectional study plus 11 patients with epidermoid supraglottic laryngeal cancer. VOCs from T3 supraglottic cancer were clustered distinctly from those of T1 and healthy subjects. Up to seven VOCs were detected differently from healthy volunteers, mainly 2-butanone and ethanol. Thus VOC-patterns of exhaled breath may discriminate patients with epidermoid head and neck cancer from healthy controls.     

Detection of volatile organic compounds (VOCs) in exhaled breath of patients with chronic obstructive pulmonary disease (COPD) by ion mobility spectrometry

COPD is a disease characterised by a chronic inflammation of the airways and a not fully reversible airway obstruction. The spirometry is considered as gold-standard to diagnose the disease and to grade its severity. In this study we used the methodology of Ion Mobility Spectometry in order to detect Volatile Organic Compounds (VOCs) in exhaled breath of patients with COPD. The purpose of this study was to investigate if the VOCs detected in patients with COPD were different from the VOCs detected in exhaled breath of healthy controls. 13 COPD patients and 33 healthy controls were included in the study. Breath samples were collected via a side-steam Teflon tube and directly measured by an ion mobility spectrometer coupled to a multi capillary column (MCC/IMS). One peak was identified only in the patients group compared to the healthy control group. Consequently, the analysis of exhaled breath could be a useful tool to diagnose COPD.     

Chromatographic analysis of VOC patterns in exhaled breath from smokers and nonsmokers

Cigarette smoking harms nearly every organ of the body and causes many diseases. The analysis of exhaled breath for exogenous and endogenous volatile organic compounds (VOCs) can provide fundamental information on active smoking and insight into the health damage that smoke is creating. Various exhaled VOCs have been reported as typical of smoking habit and recent tobacco consumption, but to date, no eligible biomarkers have been identified. Aiming to identify such potential biomarkers, in this pilot study we analyzed the chemical patterns of exhaled breath from 26 volunteers divided into groups of nonsmokers and subgroups of smokers sampled at different periods of withdrawal from smoking. Solid‐phase microextraction technique and gas chromatography/mass spectrometry methods were applied. Many breath VOCs were identified and quantified in very low concentrations (ppbv range), but only a few (toluene, pyridine, pyrrole, benzene, 2‐butanone, 2‐pentanone and 1‐methyldecyclamine) were found to be statistically significant variables by Mann–Whitney test. In our analysis, we did not consider the predictive power of individual VOCs, as well as the criterion of uniqueness for biomarkers suggests, but we used the patterns of the only statistically significant compounds. Probit prediction model based on statistical relevant VOCs‐patterns showed that assessment of smoking status is heavily time dependent. In a two‐class classifier model, it is possible to predict with high specificity and sensitivity if a subject is a smoker who respected 1 hour of abstinence from smoking (short‐term exposure to tobacco) or a smoker (labelled "blank smoker") after a night out of smoking (long‐term exposure to tobacco). On the other side, in our study "blank smokers" are more like non‐smokers so that the two classes cannot be well distinguished and the corresponding prediction results showed a good sensitivity but low selectivity.     

Noninvasive detection of colorectal cancer by analysis of exhaled breath

There has been growing interest in exhaled breath analysis for cancer screening and disease monitoring; however, limited breath biomarker information exists regarding colorectal cancer (CRC). The objective of this study was to screen for breath biomarkers of CRC. Exhaled breath was collected from 20 CRC patients and 20 healthy controls; subsequently, solid-phase microextraction–gas chromatography/mass spectrometry (SPME-GC/MS) was used to assess the exhaled volatile organic compounds (VOCs) of the study participants. The statistical methods of principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were performed to process the final data. The VOCs in the exhalations of CRC patients exhibited significant differences from the VOCs in the exhalations of healthy controls; in particular, relative to the latter exhalations, the former exhalations contain significantly higher levels of cyclohexanone, 2,2-dimethyldecane, dodecane, 4-ethyl-1-octyn-3-ol, ethylaniline, cyclooctylmethanol, trans-2-dodecen-1-ol, and 3-hydroxy-2,4,4-trimethylpentyl 2-methylpropanoate but significantly lower levels of 6-t-butyl-2,2,9,9-tetramethyl-3,5-decadien-7-yne (P?<?0.05). Analyses of breath VOCs provide a related model of CRC exhalation that could represent an effective and convenient screening method for this disease.

Human breath analysis: methods for sample collection and reduction of localized background effects

Solid-phase microextraction (SPME) was applied, in conjunction with gas chromatography–mass spectrometry, to the analysis of volatile organic compounds (VOCs) in human breath samples without requiring exhaled breath condensate collection. A new procedure, exhaled breath vapor (EBV) collection, involving the active sampling and preconcentration of a breath sample with a SPME fiber fitted inside a modified commercial breath-collection device, the RTube™, is described. Immediately after sample collection, compounds are desorbed from the SPME fiber at 250 °C in the GC-MS injector. Experiments were performed using EBV collected at −80 °C and at room temperature, and the results compared to the traditional method of collecting exhaled breath condensate at −80 °C followed by passive SPME sampling of the collected condensate. Methods are compared in terms of portability, ease-of-use, speed of analysis, and detection limits. The need for a clean air supply for the study subjects is demonstrated using several localized sources of VOC contaminants including nail polish, lemonade, and gasoline. Various simple methods to supply clean inhaled air to a subject are presented. Chemical exposures are used to demonstrate the importance of providing cleaned air (organic vapor respirator) or an external air source (tubing stretched to a separate room). These techniques allow for facile data interpretation by minimizing background contaminants. It is demonstrated herein that this active SPME breath-sampling device provides advantages in the forms of faster sample collection and data analysis, apparatus portability and avoidance of power or cooling requirements, and performance for sample collection in a contaminated environment.      

An adaptive breath sampler for use with human subjects with an impaired respiratory function

An adaptive sampler for collecting 2.5 dm(3) samples of exhaled air from human subjects with an impaired respiratory function is described. Pressure in the upper respiratory tract is continuously monitored and the data used to control an automated system to collect select portions of the expired breathing cycle onto a mixed bed Tenax(trade mark) and Carbotrap(trade mark) adsorbent trap for analysis by GC-MS. The sampling approach is intended for use in metabolomic profiling of volatiles in human breath at concentrations greater than microg m(-3). The importance of experimental reproducibility in metabolomic data is emphasised and consequently a high purity air supply is used to maintain a stable exogenous volatile organic compound profile at concentrations in the range 5 to 30 microg m(-3). The results of a 90 day stability study showed that exogenous VOCs were maintained at significantly lower levels (40 times lower for isopropyl alcohol) and with significantly higher reproducibility (80 times lower standard deviation for isopropyl alcohol) than would have been be the case if ambient air had been used. The sampling system was evaluated with healthy controls alongside subjects with chronic obstructive pulmonary disease. Subjects were able to breathe normally with control subjects observed to breathe at a rate of 9 to 17 breaths per minute, compared to 16 to 30 breaths per minute for subjects with COPD. This study presents, for the first time, observations and estimates of intra-subject breath sample reproducibility from human subjects. These reproducibility studies indicated that VOCs in exhaled breath exhibit a variety of dynamic behaviours, with some species recovered with a RSD <30%, while other species were observed to have significantly more variable concentrations, 30 to 130% RSD. The approach was also demonstrated to reliably differentiate the differences in the VOC profiles between alveolar and dead space air.