β2-兴奋剂的样品前处理与分析检测方法研究进展

许多国家禁止β2-兴奋剂用于动物饲养,但违禁药品的使用屡有发生,因此β2-兴奋剂的分析检测引起了高度关注.该文综述了β2-兴奋剂分析的样品前处理方法,包括溶剂萃取、固相萃取、固相微萃取等,并对常用的β2-兴奋剂检测方法(如色谱分析法、电化学检测法、酶联免疫法、化学发光法等)的研究进展进行了评述.     

兴奋剂中利尿剂的检测方法

利尿剂是国际奥委会医学委员会禁止使用的一类兴奋剂。该文对兴奋剂中利尿剂的检测方法进行了综述,重点阐述了样品的处理方法,HPLC－GC－MS的检测方法及各种衍生化方法。     

兴奋剂检测

在体育比赛中,有些运动员为了争取取得好名次、提高成绩,常服用一些药物,企图凭借药物作用获得荣誉.这种做法损害了体育运动的本来目的,也违反了奥林匹克公平竞赛的原则.同时,因药物的毒副作用严重地威胁着运动员的身体健康,国际奥委会(IOC)明文规定禁止运动员服用某些药物,并自1968年起在奥运会比赛中对运动员进行“滥用药物”的检测,若查出其已服用,便立即取消其参赛资格或所得名次.由于最早服用的药物多为兴奋性     

蛋白同化激素类兴奋剂质谱法检测的现状和发展

对蛋白同化激素类兴奋剂检测的发展和现状进行了评述，除目前常规的色谱／色谱联用选择离子检测方法外，强调指出了高分辨质谱和串联质谱的应用前景。     

兴奋剂与兴奋剂检测研究进展

滥用兴奋剂会遭受国际奥委会的严厉处罚 ,但仍有不少运动员为冠军及奖牌所带来的巨大利益所诱惑 ,铤而走险 ,不惜以各种方式滥用兴奋剂 ,所以兴奋剂及其检测是体育界乃至全社会关注的问题。本文主要就兴奋剂、兴奋剂检测的概念以及该领域国内外的最新研究动态和发展方向进行介绍 ,并着重探讨肽类激素rhEPO、rhGH的研究方法和对不同检测基质 (尿样、发样、血样、体液 )的检测进展     

兴奋剂──运动场上的化学幽灵

近几十年来,兴奋剂问题一直困扰着体育界,成为世人普遍关注的焦点之一。本文从化学的角度介绍了兴奋剂的种类及其检测方法。     

兴奋剂

滥用兴奋剂会遭受国际奥委会的严厉处罚,但仍有不少运动员为冠军及奖牌所诱惑,铤而走险,不惜以各种方式滥用兴奋剂,所以兴奋剂及其检测是体育界乃至全社会关注的问题.本文主要介绍兴奋剂、兴奋剂检测等.     

环境样品中多溴联苯醚分析方法的研究进展

多溴联苯醚(PBDEs)作为一类应用广泛的溴代阻燃剂,具有持久污染性、易于从被应用产品中脱离出来进入环境介质等特性,目前已对全球环境造成了严重危害.近年来针对PBDEs分析检测技术的报道愈来愈多,然而PBDEs不仅含量极低,而且所处基体复杂,因此样品前处理技术成为分析PBDEs类化合物的一个重要步骤,受到科学工作者的广泛重视.本文作者综述了近年来分析环境样品中PBDEs的样品前处理技术和分析检测方法的研究进展,为更好的发展准确、灵敏、快速的分析方法提供重要的参考.     

兴奋剂与兴奋剂检测

汉语中的兴奋剂一词原是指对中枢神经有兴奋作用的一类药物,有时又叫做中枢神经刺激剂,例如麻黄素、苯丙胺等药物。兴奋剂对应的外文词汇是dope,在Webster’s New Collegiate Dictionary中,对dope的相应注释有两条,一是“麻醉药物制品”（a narcotic preparation）,应当就是中文的毒品;     

液相色谱-电喷雾电离质谱法检测尿液中的3种合成类固醇药物

研究建立了合成类固醇药物群勃龙、四氢孕三烯炔酮和孕三烯酮的液相色谱-电喷雾质谱的检测方法。尿样经过β-葡萄糖醛酸酶酶解和叔丁基甲醚提取后,采用Zorbax SB-C18分析柱(150 mm×2.1 mm,5 μm),在流动相为pH 3.5的甲酸铵缓冲液-乙腈的条件下进行梯度洗脱,在正离子模式下检测。考察了不同的质谱条件对这类化合物检测结果的影响。建立了人尿液中合成类固醇药物的液相色谱-电喷雾电离质谱的初筛和确证方法。     

Screening for anabolic steroids in doping analysis by liquid chromatography/electrospray ion trap mass spectrometry

A fast and selective LC/MS/MS method for the screening of four anabolic steroids in human urine has been developed and validated. Liquid-liquid extraction with diethyl ether was applied after enzymatic hydrolysis. Analyses were performed on an ion trap mass spectrometer equipped with electrospray ionisation. MS/MS was applied for all compounds. The analytical run time was 11 min. The LOD for all compounds varied between 1 and 10 ng/mL. Left-over A samples, which were declared positive by GC/MS for the presence of 3'-hydroxystanozolol, were assessed using the described method.     

Ionization of anabolic steroids by adduct formation in liquid chromatography electrospray mass spectrometry

The ionization of 46 anabolic steroids has been studied. The absence of basic or acidic moieties in most of these analytes makes their direct ionization as [M + H]+ by atmospheric pressure interfaces difficult. The formation of adducts with different components of the mobile phase has been found to be an efficient way to ionize anabolic steroids by electrospray. Different mobile phases using methanol (MeOH) or acetonitrile as organic solvent and HCOOH, Na+ or NH4+ as additives have been tested to favor the adduct formation. A direct correlation between the chemical structure of the anabolic steroid and the possibility to ionize it in a particular chromatographic condition has been found. According to their ionization, anabolic steroids can be divided into seven different groups depending on both the nature and the relative position of their functional groups. The formation of different adducts such as [M + Na + MeOH]+ or [M + H + CH3 CN - H2O]+ is required in order to ionize some of these groups and the optimal mobile phase composition for each group of anabolic steroids is proposed. Despite the ionization limitations due to their chemical structure, most of tested anabolic steroids could be ionized using the adduct formation approach.     

Determination of anabolic androgenic steroids as imidazole carbamate derivatives in human urine using liquid chromatography–tandem mass spectrometry

Anabolic androgenic steroids are widely abused substances in sports doping. Their detection present limitations regarding the use of soft ion sources such as electrospray or atmospheric pressure chemical ionization by liquid chromatography–tandem mass spectrometry. In the current study, a novel derivatization method was developed for the ionization enhancement of selected anabolic androgenic steroids. The proposed method aims at the introduction of an easily ionizable moiety into the steroid molecule by converting the hydroxyl groups into imidazole carbamates using 1,1′‐carbonyldiimidazole as derivatization reagent. The proposed method was applied to water and urine samples spiked with exogenous anabolic androgenic steroids in various concentration levels. Steroid imidazole carbamate derivatives have shown intensive [M+H]⁺ signals under electrospray ionization and common fragmentation patterns in tandem mass spectrometry mode with [M‐CO₂+H]⁺ and [M‐ΙmCO₂+H]⁺ as major ions with low collision energy. The obtained results showed that the majority of steroids were detectable at concentrations equal or lower to their minimum required performance level according to the World Anti‐Doping Agency technical document. The proposed method is sensitive with a preparation procedure that could be easily applied to the analysis of doping control samples.     

Analysis of anabolic steroids in hair by GC/MS/MS

A simple and sensitive gas chromatography/tandem mass spectrometry (GC/MS/MS) method is described for the detection of anabolic steroids, usually found in keratin matrix at very low concentrations. Hair samples from seven athletes who spontaneously reported their abuse of anabolic steroids, and in a single case cocaine, were analyzed for methyltestosterone, nandrolone, boldenone, fluoxymesterolone, cocaine and its metabolite benzoylecgonine. Anabolic steroids were determinate by digestion of hair samples in 1 m NaOH for 15 min at 95 degrees C. After cooling, samples were purificated by solid-phase and liquid-liquid extraction, then anabolic steroids were converted to their trimethylsilyl derivative and finally analyzed by GC/MS/MS. For detection of cocaine and benzoylecgonine, hair samples were extracted with methanol in an ultrasonic bath for 2 h at 56 degrees C then overnight in a thermostatic bath at the same temperature. After the incubation, methanol was evaporated to dryness, and benzoylecgonine was converted to its trimethylsilyl derivative prior of GC/MS/MS analysis. Results obtained are in agreement with the athletes' reports, confirming that hair is a valid biological matrix to establish long-term intake of drugs.     

高效液相色谱法同时测定血浆中的10种蛋白同化激素

建立了一种用于10种蛋白同化激素的同时分离检测的高效液相色谱法。根据被分析物的性质,以C18反相色谱柱为分离柱,以乙腈和水为流动相,采用梯度洗脱方式,并在194～290 nm的范围内快速调节检测波长,使各物质均在最大吸收波长处被检出。在优化的条件下,10种被测组分在10 min内实现了快速的基线分离,检出限在0.01～0.10 μg/mL范围内。在兔血浆中进行加标回收率测定,10种被测组分的加标回收率为70.3%～120%。选取美雄醇为代表进行实际动物实验,成功检测到耳脉注射美雄醇后兔血浆内的美雄醇成分。实验结果表明该方法可行,快速简便,准确可靠。     

液相色谱-质谱联用在兴奋剂检测中的应用及进展

液相色谱-质谱联用技术已越来越广泛地应用在兴奋剂的检测中,其中包括对各类小分子兴奋剂和肽类激素等的检测。本文综述了近年来液相色谱-质谱联用在兴奋剂检测中的筛选、确证和定量方面的应用及进展情况,并讨论了相关的检测标准。     

External calibration in gas chromatography-combustion-isotope ratio mass spectrometry measurements of endogenous androgenic anabolic steroids in sports doping control

An alternative calibration procedure for the Gas Chromatography–Combustion–Isotope Ratio Mass Spectrometry (GC–C–IRMS) measurements of the World Antidoping Agency (WADA) Accredited Laboratories is presented. To alleviate the need for externally calibrated CO₂ gas for GC–C–IRMS analysis of urinary steroid metabolites, calibration using an external standard mixture solution of steroids with certified isotopic composition was investigated. The reference steroids of the calibration mixture and routine samples underwent identical instrumental processes. The calibration standards bracketed the entire range of the relevant δ¹³C values for the endogenous and exogenous steroids as well as their chromatographic retention times. The certified δ¹³C values of the reference calibrators were plotted in relation to measured m/z¹³CO₂/¹²CO₂ (i.e. R(45/44)) mass spectrometric signals of each calibrator. δ¹³C values of the sample steroids were calculated from the least squares fit through the calibration curve. The effect of the external calibration on δ¹³C values, using the same calibration standards and set of urine samples but different brands of GC–C–IRMS instruments, was assessed by an interlaboratory study in the WADA Accredited Laboratories of Sydney, Australia and Athens, Greece. Relative correspondence between the laboratories for determination of androsterone, etiocholanolone, 5β-androstane-3α,17β-diacetate, and pregnanediacetate means were SD(δ¹³C) = 0.12‰, 0.58‰, −0.34‰, and −0.40‰, respectively. These data demonstrate that accurate intralaboratory external calibration with certified steroids provided by United States Antidoping Agency (USADA) and without external CO₂ calibration is feasible and directly applicable to the WADA Accredited Laboratories for the harmonization of the GC–C–IRMS measurements.     

Characterization of chemically modified steroids for doping control purposes by electrospray ionization tandem mass spectrometry

The discovery of the designer steroid tetrahydrogestrinone (THG) in elite athletes' doping control samples in 2003 demonstrated the availability of steroid derivatives prepared solely for doping purposes. Modern mass spectrometers utilizing electrospray ionization and collisionally activated dissociation (CAD) of analytes allow the structural characterization of steroids and their derivatization sites by the elucidation of fragmentation behaviors. A total of 21 steroids comprising either a 4,9,11-triene, a 3-keto-4-ene or a 3-keto-1-ene nucleus were investigated regarding their dissociation pathways, deuterated analogues were synthesized and fragmentation routes were postulated, permitting the identification of steroidal structures and modifications. Compounds based on a 4,9,11-triene steroid with an ethyl residue at C-13 (gestrinone analogues) generate abundant fragment ions at m/z 241 and 199, whereas the substitution of the C-13 ethyl group by a methyl residue (trenbolone analogues) results in a shift of m/z 241 to 227. Substances related to testosterone with a 3-keto-4-ene structure give rise to abundant fragment ions at m/z 109 and 97 whereas steroids with a 3-keto-1-ene nucleus eliminate the A-ring including the carbons C-1-C-4, in addition to C-19 that is proposed to migrate from C-10 to C-1 under CAD conditions.     

Mass spectrometric behavior of anabolic androgenic steroids using gas chromatography coupled to atmospheric pressure chemical ionization source. Part I: Ionization

The detection of anabolic androgenic steroids (AAS) is one of the most important topics in doping control analysis. Gas chromatography coupled to (tandem) mass spectrometry (GC–MS(/MS)) with electron ionization and liquid chromatography coupled to tandem mass spectrometry have been traditionally applied for this purpose. However, both approaches still have important limitations, and, therefore, detection of all AAS is currently afforded by the combination of these strategies. Alternative ionization techniques can minimize these drawbacks and help in the implementation of a single method for the detection of AAS. In the present work, a new atmospheric pressure chemical ionization (APCI) source commercialized for gas chromatography coupled to a quadrupole time‐of‐flight analyzer has been tested to evaluate the ionization of 60 model AAS. Underivatized and trimethylsylil (TMS)‐derivatized compounds have been investigated. The use of GC–APCI–MS allowed for the ionization of all AAS assayed irrespective of their structure. The presence of water in the source as modifier promoted the formation of protonated molecules ([M+H]⁺), becoming the base peak of the spectrum for the majority of studied compounds. Under these conditions, [M+H]⁺, [M+H‐H₂O]⁺ and [M+H‐2·H₂O]⁺ for underivatized AAS and [M+H]⁺, [M+H‐TMSOH]⁺ and [M+H‐2·TMSOH]⁺ for TMS‐derivatized AAS were observed as main ions in the spectra. The formed ions preserve the intact steroid skeleton, and, therefore, they might be used as specific precursors in MS/MS‐based methods. Additionally, a relationship between the relative abundance of these ions and the AAS structure has been established. This relationship might be useful in the structural elucidation of unknown metabolites. Copyright © 2014 John Wiley & Sons, Ltd.