液相色谱-串联质谱法测定头发中10种蛋白同化激素

建立了同时测定头发中10种蛋白同化激素液相色谱-串联质谱(LC/MS/MS)分析方法。头发样品经NaOH消解、戊烷液液提取后,用反相液相色谱分离,电喷雾正离子源进行离子化,用多反应监测方式(MRM)对这10种蛋白同化激素的母离子及子离子进行监测,三重四极杆质谱测定。10种蛋白同化激素的检出限为1~20ng/g;相对标准偏差(RSD)为1.72%~13.77%;回收率为38.20%~110.38%;线性回归系数(R2)为0.9958~0.9999。本方法简便快速、灵敏度高、专属性强,可满足在兴奋剂检测或毒物分析中对毛发中蛋白同化激素测定的要求。     

气质联用法同时测定猪尿中的20种同化激素

采用安捷仑5975GC/MS对猪尿中20种同化激素进行同时检测.对于10.0 g尿液样品,采用β-葡糖苷酸酶(来自Helix pomatia)对提取物水解后,加入过饱和NaCl,经乙腈提取,用过C18小柱和NH2小柱净化后七氟丁酸酐对其进行衍生,最后利用GC/MS进行检测.采用外标法定量,以S/N≥3确定的,检出限均能达到0.25 μg/kg.通过对3个添加水平的回收实验表明,平均回收率为77.0%～111.0%.     

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。     

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。     

高效液相色谱-串联质谱法检测配合饲料中11种蛋白同化激素含量

建立了快速分析配合饲料中睾酮、甲基睾酮、勃地龙、美雄酮、雄烯二酮、脱氢异雄酮、诺龙、丙酸诺龙、司坦唑醇、美伦孕酮、黄体酮11种蛋白同化激素的高效液相色谱-串联质谱(HPLC-MS/MS)法。样品采用乙腈提取,经PSA粉净化后上机测定。采用Phenomenex C₁₈(100 mm×2.1 mm,2.6μm)色谱柱,以0.01%甲酸溶液-乙腈作为流动相进行梯度洗脱,电喷雾离子源正离子模式检测,同位素内标法定量。结果表明,11种蛋白同化激素的线性范围为1～100 ng/mL,相关系数均为0.999,检出限为20μg/kg,定量下限为50μg/kg。50、250、500μg/kg加标水平下,鸡配合饲料中各蛋白同化激素的回收率为94.5%～111%,日内相对标准偏差(RSD)和日间RSD均不大于13%;猪配合饲料中各蛋白同化激素的回收率为90.1%～109%,日内RSD不大于9.0%,日间RSD不大于8.8%。实际样品中检出睾酮和勃地龙,含量分别为9.09～14.68 mg/kg和1.22～1.84 mg/kg。该方法可为饲料中蛋白同化激素的滥用监管提供技术支撑。     

动物尿液中15种甾类同化激素的液相色谱-串联质谱同时测定

采用液相色谱-电喷雾串联质谱仪（LC-ESIMS/MS）,在多反应监测（MRM）模式下建立了动物尿液中15种甾类同化激素（睾酮、孕酮、诺龙、甲基睾酮、丙酸睾酮、丙酸诺龙、苯丙酸诺龙、大力补、勃地龙、群勃龙、康力龙、醋酸群勃龙、甲炔诺酮、甲羟孕酮、醋酸甲羟孕酮）的快速确证测定方法。试样经酶解处理后,过C18固相萃取柱净化,氮吹至干,残余物用0.50 mL乙腈-水（体积比1∶1）溶解后测定。采用正离子扫描方式进行仪器方法学研究,确定监测离子对,进行MRM模式定性定量分析。该方法的检出限（LOD）为0.2～0.5μg/L,定量下限（LOQ）为0.5～1.0μg/L;在2.0～200.0μg/L范围内线性关系良好,相关系数均大于0.998。在1.0、5.0μg/L的添加水平上,上述15种激素的平均回收率为59%～118%,相对标准偏差为1.0%～11.2%。该法操作简单,灵敏度高,可用于动物尿液中15种甾类同化激素的测定。     

气相色谱-质谱法检测动物肌肉组织中残留的甾类同化激素

建立了不同动物肌肉中甾类同化激素残留的气相色谱/质谱（GC/MS）检测方法。样品经酶解、组织捣碎和超声提取后,用叔丁基甲醚萃取和反相固相萃取柱净化,进一步衍生后进行GC/MS分析,分别测定雄激素（表睾酮（ETS）、丙酸睾酮（PTS）、19-去甲基睾酮（17β-NT）、甲基睾酮（MTS）)和雌激素（雌二醇（17β-ES）、雌三醇（EST）、炔雌醇（EES）、雌酮（ESN）、苯甲酸雌二醇（BES)）。实验结果表明,ETS、17β-NT、MTS、PTS、BES、ESN、17β-ES、EES和EST的定量检测限为1.0~2.0 μg/kg。在2.0 μg/kg的定量检测低限添加水平,上述9种激素的平均回收率为62.5%~80.5%,相对标准偏差为12.5%~26.8%。该法可实现对样品进行灵敏、准确的定性定量分析。     

动物肌肉组织中甾类同化激素多组分残留的液相色谱-质谱检测方法

建立了不同动物肌肉中甾类同化激素(表睾酮、丙酸睾酮、19-去甲基睾酮、甲基睾酮、孕酮、甲羟孕酮、雌二醇、雌三醇、炔雌醇、雌酮)多残留量的LC/MS/MS确证方法。样品加醋酸缓冲溶液均质,加酶溶液酶解后,再加甲醇超声提取,用叔丁基甲醚液-液萃取至少2次,之后经反相固相萃取柱净化,以乙腈-水为流动相,经C18柱分离后进行LC/MS/MS多反应监测模式下的定性及定量分析。其中雄激素,孕激素采用正离子扫描,雌激素则采用负离子扫描。17beta-NT、MTS、ETS、MED、PG、PTS、EST、17beta-ES、EES和ESN的定量检出限为0.5~1.0μg/kg。在1.0μg/kg的定量检出限添加水平,上述10种激素的平均回收率为55%~77%;相对标准偏差为7.1%~35%。可实现样本灵敏、准确的定性定量分析。     

多重机制杂质吸附萃取净化-快速液相色谱-串联质谱法测定鱼组织中11种同化激素

建立了准确、灵敏的鱼组织中11种同化激素(勃地酮、雄烯二酮、诺龙、美雄酮、甲睾酮、睾酮、醋酸睾酮、群勃龙、丙酸睾酮、康力龙、氟甲睾酮)的多重机制杂质吸附萃取净化-快速液相色谱-串联质谱的分析方法。鱼组织均质样品经甲醇提取后,在上清液中加入一定量的C18固体吸附剂、中性氧化铝吸附剂和氨基功能化纳米吸附剂实现快速净化。采用Shim-Pack XR-ODSII色谱柱(100 mm×2.0 mm, 2.2 μm)分离,以乙腈(含0.1%甲酸)和水(含0.1%甲酸)为流动相进行梯度洗脱,电喷雾正离子多反应监测(MRM)模式下检测,外标法定量。结果表明,11种目标化合物在线性范围内具有良好的线性关系,相关系数大于0.999,其在鱼组织中的检出限(S/N＞3)为0.03～0.4 μg/kg,定量限(S/N＞10)为0.1～1.5 μg/kg,平均回收率为80.9%～98.1%,相对标准偏差(RSD)为5.2%～11.5%。该方法简便、快速、准确,可用于鱼组织中同化激素的定性、定量监测。     

头发中内源性类固醇激素的气相色谱-串联质谱分析

建立了建康人头发中内源性类固醇兴奋剂睾酮、表睾酮、雄酮、苯胆烷醇酮和脱氢表雄酮的气相色谱-串联质谱(GC-MS/MS)分析方法。头发经碱水解后,以乙醚提取,经衍生化后采用GC-MS/MS的多反应监测模式(MRM)分析。方法的线性关系良好,检出限达0.1～0.2 pg/mg;提取回收率为74.6%～104.5%;日内测定的准确度为90.1%～113.7%,日内及日间测定的精密度均小于17.5%。应用所建立的方法测定了80例中国健康人头发中睾酮、表睾酮、雄酮、苯胆烷醇酮和脱氢表雄酮的生理水平,为内源性类固醇兴奋剂滥用的判断提供了方法和基础数据。     

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.     

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.     

Correlation of product ion profiles with molecular structures of androgenic and anabolic steroids in ESI MS/MS

Androgenic and anabolic steroids (AASs) are a class of chemical substances closely related to testosterone in molecular structure. They can be abused to enhance performances in human and equine athletes, and are banned by the sports authorities. To assist with method development for doping analyses of AASs, investigations were conducted to correlate their product ion profiles with the molecular structures. Although very similar in chemical structure, AASs generated noticeably different product ion profiles from collision‐induced dissociation (CID). On the basis of both outlines of the product ion profiles and molecular structures, AASs studied were classified into six subclasses. In each subclass, the product ion profiles were identical or similar. However, the product ion profiles in one subclass were remarkably different from those in another. The classification reveals that the position and number of double bond(s) in conjugation with the 3‐carbonyl group in the molecular structure of an AAS have significant effects on product ion profile. The presence or absence of the 19‐methyl group in an AAS also has a remarkable influence on its product ion profile. A substitution in the A‐, B‐ or D‐ring of an AAS may cause a shift in mass value of the product ions. The correlation of product ion profiles with molecular structures of AASs has the implication that each AAS can be characterized by a combination of its [M + H]⁺ ion and product ion profile and as a result be identified with specificity. The classified product ion pattern may be useful in the identification of unknown AASs. Copyright © 2010 John Wiley & Sons, Ltd.     

固相萃取-气相色谱-质谱法检测尿液中乙基葡萄糖醛酸苷

建立了固相萃取(SPE)-气相色谱-质谱(GC-MS)检测尿液样本中乙基葡萄糖醛酸苷(EtG)的方法.1 mL尿液样本经100μL 3 mol/L盐酸去蛋白后,通过SPE法提取上清液中的目标物质及内标,提取物经衍生化后,采用GC-MS检测,选择离子模式(SIM)扫描,内标法定量分析.该方法在0.1～3.2 mg/L范围...     

Potential of atmospheric pressure chemical ionization source in gas chromatography tandem mass spectrometry for the screening of urinary exogenous androgenic anabolic steroids

The atmospheric pressure chemical ionization (APCI) source for gas chromatography-mass spectrometry analysis has been evaluated for the screening of 16 exogenous androgenic anabolic steroids (AAS) in urine. The sample treatment is based on the strategy currently applied in doping control laboratories i.e. enzymatic hydrolysis, liquid–liquid extraction (LLE) and derivatization to form the trimethylsilyl ether-trimethylsilyl enol ether (TMS) derivatives. These TMS derivatives are then analyzed by gas chromatography tandem mass spectrometry using a triple quadrupole instrument (GC-QqQ MS/MS) under selected reaction monitoring (SRM) mode. The APCI promotes soft ionization with very little fragmentation resulting, in most cases, in abundant [M + H]⁺ or [M + H-2TMSOH]⁺ ions, which can be chosen as precursor ions for the SRM transitions, improving in this way the selectivity and sensitivity of the method. Specificity of the transitions is also of great relevance, as the presence of endogenous compounds can affect the measurements when using the most abundant ions. The method has been qualitatively validated by spiking six different urine samples at two concentration levels each. Precision was generally satisfactory with RSD values below 25 and 15% at the low and high concentration level, respectively. Most the limits of detection (LOD) were below 0.5 ng mL⁻¹. Validation results were compared with the commonly used method based on the electron ionization (EI) source. EI analysis was found to be slightly more repeatable whereas lower LODs were found for APCI. In addition, the applicability of the developed method has been tested in samples collected after the administration of 4-chloromethandienone. The highest sensitivity of the APCI method for this compound, allowed to increase the period in which its administration can be detected.     

基于气相色谱-质谱联用与液相色谱-质谱联用的非靶向代谢组学用于3类茶叶中化学成分分析（英文）

茶叶中的化学成分是构成其风味特征的物质基础。本研究建立了基于气相色谱-质谱联用和液相色谱-质谱联用的非靶向代谢组学方法。完成预处理及分析条件优化后,使用标准样品考察方法的线性、回收率及重复性,结果表明方法整体稳定且结果可靠。将该方法用于绿茶、乌龙茶和红茶中化学成分的分析。通过超声辅助溶剂提取及气相色谱-质谱联用分析共获得1812个特征峰,而使用加热溶剂提取及液相色谱-质谱联用分析可获得2608个特征峰。结合保留规律及质谱数据库,共定性173种化合物(109种随后经标准样品验证),其中只有9种化合物在上述两类分析中被同时检出,表明方法互补性良好。3类茶叶数据的偏最小二乘判别分析结果表明,三者间存在显著差异。结合模型的变量重要因子(VIP)与非参数检验共筛选出90种化合物,其中包含儿茶素、氨基酸、糖、有机酸和黄酮苷类等众多与茶叶滋味密切相关的化学成分。     

张裕XO级白兰地挥发性成分的提取分离与鉴定

建立了一套对白兰地挥发性物质进行预处理的方法,并采用气相色谱-质谱法(GC-MS)较全面地鉴定了白兰地的挥发性成分。实验先采用液-液萃取方法提取张裕XO级白兰地的挥发性成分,然后将酸性成分与碱性和中性成分分离,再采用柱色谱分离手段将其分离为若干个级分并浓缩,采用气相色谱-质谱、标准品比对、保留指数(RI)值比较等方法对分离得到的各级分中的成分进行了鉴定,在白兰地中共鉴定出302种挥发性成分,包括醇30种、醛酮类35种、酸类20种、酯类104种、苯同系物及其衍生物24种、酚类14种、缩醛14种、呋喃类16种、萜烯类22种和其他物质23种。结果表明,采用这套预处理方法能将白兰地的挥发性成分较有效地分组和浓缩。     

气相色谱-质谱法在人体气味分析中的应用进展

气相色谱-质谱法（GC-MS）是目前分析人体气味的主要技术手段，其不但可以分析人体气味的组成成分，还可以比较不同个体气味之间的差异。该文首先介绍了人体气味的提取和样品前处理的手段，随后对GC-MS在人体气味的采集及储存、成分分析及差异性研究、用于个体识别和健康表征等方面进行了综述，最后对GC-MS在人体气味分析中的发展趋势进行了展望。     

Combination of liquid chromatography/tandem mass spectrometry and gas chromatography/mass spectrometry for the detection of 21 anabolic steroid residues in bovine urine

For the detection of anabolic steroid residues in bovine urine, a highly sensitive liquid chromatographic/electrospray ionization tandem mass spectrometric (LC/ESI-MS/MS) method was developed using both positive and negative ionization. For four compounds the ESI mode was not sensitive enough and gas chromatographic/mass spectrometric GC/MS detection was therefore still necessary as a complementary method. The sample clean-up consisted of solid-phase extraction (SPE) on a C(18) column followed by enzymatic hydrolysis and a second solid-phase extraction on a combination of a C(18) and a NH(2) column. After this last SPE clean-up, the eluate was split into two equal aliquots. One aliquot was further purified and after derivatization used for GC/MS analysis. The other aliquot was analyzed with LC/MS/MS in both ESI+ and ESI- modes. The method was validated according to the European Commission Decision 2002/657/EC. Decision limits (CCalpha) were between 0.16 and 1 ng ml(-1) for the compounds detected with the LC/MS/MS method. The developed method is used in routine analysis in our laboratory.     

基于稀疏模型的气相色谱-质谱数据解析及其在严重重叠峰解析中的应用

提出一种气相色谱-质谱（GC-MS）数据解析算法。以色谱峰顶点处的质谱作为待测谱,在谱库中检索一定量相关参考谱,求解关于各纯组分色谱响应值的方程。质谱检索采取分步策略,先利用质谱碎片规律建立高速索引进行粗选,然后使用强峰高概率出峰准则和耐挤压性准则排除无关质谱。为求解色谱响应值方程,提出基于稀疏模型的回归算法,相比传统算法,稀疏算法利于提取待测谱的主要结构,避免"过拟合"。实验结果表明,该质谱检索算法具有较高的精度和规模较小的剩余参考谱集,而所提稀疏模型算法可有效解析严重重叠峰。该算法可作为GC-MS重叠峰解析,特别是严重重叠峰解析的一种有效解决方案。