微乳液毛细管电动色谱-场放大富集法灵敏分析尿液中的9种麻醉剂类药物

建立了一种微乳液毛细管电动色谱(MEEKC)-场放大富集(FASI)分析尿液中多种麻醉剂(吗啡、可待因、纳洛酮、海洛因、蒂巴因、可卡因、哌替啶、芬太尼、美沙酮)的方法。考察了微乳液组成、分离电压等因素的影响,得到的最佳微乳液组成(质量分数)为0.6%十二烷基硫酸钠、1.2%正丁醇、0.6%乙酸乙酯和97.6% 10 mmol/L硼砂缓冲液(pH 9.5);分离电压为25 kV。在上述微乳体系中,9种化合物在15 min内得到了基线分离。采用场放大在线富集技术提高了分析灵敏度,检出限(S/N=3)低至0.3 μg/L。模拟尿样样品中9种麻醉剂的加标回收率介于79.4%～119.9%之间,日内相对标准偏差小于5.5%。将该方法应用于美沙酮大鼠体外代谢样品的测定,结果令人满意。     

Pressure-assisted capillary electrochromatography with electrospray ionization-mass spectrometry based on silica-based monolithic column for rapid analysis of narcotics

A pressure-assisted CEC (pCEC) with ESI-MS based on silica-based monolithic column was developed for rapid analysis of narcotics. Combining the extremely high permeability and separation efficiency of silica-based monolithic column with the high selectivity and sensitivity of pCEC-ESI-MS, the developed system exhibited its prominent advantages in separation and detection. A systematic investigation of the pCEC separation and ESI-MS detection parameters was performed. Experiment results showed that the optimized separation efficiency could be obtained at 8 bar assisted pressure with 25 kV separation voltage, using the solution containing 65% ACN v/v and 20 mmol/L ammonium acetate with pH 6.0 as running buffer. 3 microL/min of sheath liquid was considered as the optimized flow rate since it could provide the maximum signal intensity. Under the optimum conditions, the tested five narcotics could be completely separated within 10 min with the detection limit in the range of 2.0-80 nmol/L. The proposed method has been successfully used for detection of narcotics in real urine samples.     

Determination of antibiotics in feedstuff samples by microemulsion electrokinetic chromatography using fullerene as additive

A microemulsion electrokinetic chromatography method was developed for the simultaneous detection and quantification of ciprofloxacin, norfloxacin, sulfamethoxazole, tetracycline, and oxytetracycline in feedstuff samples. The BGE composition was optimized by applying a Taguchi parameter design and consisted of phosphate 30 mmol/L, Tween‐80 0.01 mmol/L, and fullerene 3 μmol/L, and adjusted to pH 8.0; the addition of surfactant and fullerene modifies the mobility of the analytes improving their resolution. Theoretical studies showed π‐π and van der Waals interactions between antibiotic molecules and fullerene used as a pseudostationary phase. Under optimal conditions, limits of detection ranged from 0.7 to 1.5 μg/g; the analytes were separated in less than 6 min. The methodology proposed is useful for controlling and monitoring antibiotic residues in feedstuff samples.     

Pressurized liquid extraction-capillary electrophoresis-mass spectrometry for the analysis of polar antioxidants in rosemary extracts

A method based on capillary electrophoresis-electrospray-mass spectrometry (CE-ESI-MS) was developed to qualitatively characterize natural antioxidants from rosemary (Rosmarinus officinalis L.) in different fractions obtained by pressurized liquid extraction (PLE) using subcritical water. The parameters of CE-ESI-MS were adjusted allowing the separation and characterization of different compounds from rosemary in the PLE fractions. These parameters for CE are kind, pH and concentration of the separation buffer, parameters for ESI-MS are dry gas temperature and flow, nebulizing gas pressure, and make-up flow. The following analytical conditions were found most favorable: aqueous CE buffer (40 mM ammonium acetate/ammonium hydroxide, pH 9); sheath liquid containing 2-propanol-water (60:40, v/v) and 0.1% (v/v) triethylamine at a flow rate of 0.24 mL/h; drying gas flow rate equal to 7 L/min at 350 degrees C, nebulizing gas pressure of 13.8 kPa (2 psi), using a compound stability of 50%. Different antioxidant compounds (e.g., rosmarinic acid and carnosic acid) could be detected in the rosemary extracts by CE-ESI-MS without any additional treatment, enabling the determination of variations in the extract composition caused by the different PLE conditions (i.e., 60 and 100 degrees C). The results provide complementary information to HPLC analysis.     

Trace determination of cannabinoids and opiates in wastewater and surface waters by ultra-performance liquid chromatography-tandem mass spectrometry

A fast and reliable method using solid-phase extraction and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) has been developed for the simultaneous detection, identification and quantification of several central nervous system depressor drugs of abuse such as cannabinoids (Delta9-tetrahydrocannabinol, THC) and opiates (morphine, codeine, heroin, methadone, fentanyl) and their metabolites in water samples. Compounds were extracted from water by using Oasis HLB cartridges. After SPE enrichment, the selected depressor drugs, under UPLC optimized conditions, were separated in less than 8 min. Electrospray (ESI) tandem MS in positive ion mode and selected reaction monitoring was used for quantification. ESI-MS/MS conditions such as capillary and cone voltages, source and desolvation temperatures and cone and desolvation gas flow rates have been optimized and MS and MS/MS spectra of the studied compounds were obtained. At the working conditions four identification points were obtained as required by European Union guidelines for analysis by LC-MS/MS. Quality parameters (intra-day and inter-day precisions) for each analyte have been established in three different matrixes (purified, surface and waste waters). Recoveries were generally higher than 70% and instrumental quantification limits and limits of quantification were in the low pg and ng/l range, respectively. Finally, the method has been applied to the analysis of influent and effluents wastewaters and natural water samples from Catalonia (NE Spain) where the presence of several opiates such as morphine, codeine, norcodeine 2-ethylene-1,5-dimethyl-3,3-diphenylpyrrolidine and methadone and cannnabinoids such as THC and 11-nor-carboxy-Delta9-tetrahydrocannabinol has been demonstrated.     

胶束电动毛细管色谱法检测红曲米中的莫纳可林K

建立了测定红曲米中莫纳可林K含量的胶束电动毛细管色谱(MEKC)方法。考察了运行缓冲液的种类、pH及其浓度、有机添加剂、十二烷基硫酸钠(SDS)的浓度和分离电压等实验条件对电泳分离效果及检测灵敏度的影响。在优化的实验条件下,以20 mmol/L硼砂(pH 10.6,含10%(体积分数)乙醇和40 mmol/L SDS)作为缓冲溶液,莫纳可林K能在23 min内实现很好的基线分离,线性范围为5.00～100.00 mg/L,线性相关系数为0.9976,检出限(以信噪比(S/N)为3计)为0.13 mg/L,加标回收率为98.5%～99.5%。精密度和稳定性试验中,峰面积和迁移时间的相对标准偏差均小于3%,表明重复性良好。该方法简便、快速、灵敏,可用于红曲米中莫纳可林K含量的测定。     

亚微米C18键合硅胶固定相加压毛细管电色谱法同时拆分3种手性三唑类农药

采用改良Stöber法制备420 nm亚微米单分散二氧化硅微球,采用C18硅烷化修饰后装填成毛细管色谱柱。采用该色谱柱,在加压毛细管电色谱平台上成功地实现了3对手性三唑类农药烯效唑、烯唑醇和丙环唑的同时拆分和分离。考察了各因素对手性分离效果的影响,优化后的色谱条件为：流动相为乙腈-20 mmol/L磷酸盐缓冲液（pH=6.8）（45：55,v/v）,其中缓冲液中含20 mmol/L羟丙基-γ-环糊精（HP-γ-CD）;泵流速为0.04 mL/min;施加电压-9.4 kV;检测波长220 nm。在上述条件下,烯效唑、烯唑醇和丙环唑3种对映体同时得到拆分和分离,相邻两峰之间的分离度依次为4.20、12.9、4.41、4.09、1.70,分离时间仅为12 min,柱效最高达到310000 plates/m。该研究为手性三唑类农药的同时分离提供了新的分离分析思路。     

Fast determination of codeine,orphenadrine, promethazine,scopolamine, tramadol,and paracetamol in pharmaceutical formulations by capillary electrophoresis

Paracetamol is an active ingredient commonly found in pharmaceutical formulations in combination with one of the following compounds: codeine, orphenadrine, promethazine, scopolamine, and tramadol. In this work, we propose a unique analytical method for determination of these active ingredients in pharmaceutical samples. The method is based on capillary electrophoresis with capacitively coupled contactless conductivity detection. The separation was achieved on a fused silica capillary (50 cm total length, 40 cm effective length, and 50 μm id) using an optimized background electrolyte composed of 20 mmol/L β‐alanine/4 mmol/L sodium chloride/4 μmol/L sodium hydroxide (pH 9.6). Each sample can be analyzed in a single run (≤2 min) and the limits of detection were 2.5, 0.62, 0.63, 2.5, 15, and 1.6 μmol/L for scopolamine, tramadol, orphenadrine, promethazine, codeine, and paracetamol, respectively. Recovery values for spiked samples were between 94 and 104%.     

Baseline separation of α and β‐acids homologues and isomers in hop (Humulus lupulus L.) by CD‐MEKC‐UV

An alternative method for simultaneous baseline separation of α and β‐acids homologues and isomers in hop by CD‐MEKC with UV detection was proposed. The optimized background electrolyte was composed of 30 mmol/L sodium tetraborate solution, 45 mmol/L sodium dodecyl sulfate, 20 mmol/L β‐cyclodextrin and 10% v/v acetonitrile. The instrumental conditions were evaluated by using a 3³ Box‐Benhken experimental design. In order to demonstrate the applicability of the method, 21 hop samples from different varieties were analyzed. The repeatability intra‐ and interday tests were performed and relative standard deviations lower than 7% for area and migration times were observed. The present method comprehended 8 min analysis time and revealed to be faster and more efficient when compared to previous reports from literature.     

pCEC coupling with ESI‐MS for the analysis of β2‐agonists and narcotics using a poly‐(1‐hexadecene‐co‐TMPTMA) monolithic column

A pressure‐assisted CEC with ESI‐MS based on poly(1‐hexadecene‐co‐trimethylolpropane trimethacrylate) monolithic column for rapid analysis of two β₂‐agonists and three narcotics was established in this article. After the organic polymer‐based monolithic column was prepared by an in‐situ polymerization procedure, a systematic investigation of the pressure‐assisted CEC separation and ESI‐MS detection parameters was performed. Baseline separation of the studied analytes could be obtained using the solution containing 75% ACN v/v and 20 mmol/L ammonium acetate with pH 8.0 as running buffer, when applying separation voltage of 20 kV and assisted pressure of 5 bar. Under the optimized conditions, two β₂‐agonists and three narcotics could be completely resolved and accurately determined within 15 min. Finally, the proposed method was successfully used for real urine samples detection.     

阳离子选择性耗尽进样-胶束电动色谱法对可非糖浆中盐酸异丙嗪与磷酸可待因的测定

在胶束电动色谱法的基础上,联用阳离子选择性耗尽进样技术,对盐酸异丙嗪和磷酸可待因同时测定的方法进行了研究。考察了pH值、有机溶剂、SDS浓度、进样时间、进样电压等实验条件对分离效果的影响。最佳实验条件为:缓冲体系16%乙腈+80 mmol/L SDS+20 mmol/L NaH2PO4(pH2.4),分离电压为-18 kV,测量波长214 nm,萃取液pH2.4,进样电压10 kV,进样时间100 s。在优化实验条件下,两种物质在8 min内出峰,峰面积RSD不大于4.6%。盐酸异丙嗪、磷酸可待因的线性范围分别为0.50~81.3、0.78~62.5μg/L,检出限分别为0.16、0.12μg/L,相关系数分别为0.998 9、0.998 8。将方法用于可非糖浆中盐酸异丙嗪与磷酸可待因的测定,回收率为96%~106%。     

High-throughput simultaneous analysis of buprenorphine, methadone, cocaine, opiates, nicotine, and metabolites in oral fluid by liquid chromatography tandem mass spectrometry

A method for simultaneous determination of buprenorphine (BUP), norbuprenorphine (NBUP), methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), cocaine, benzoylecgonine (BE), ecgonine methyl ester (EME), anhydroecgonine methyl ester (AEME), morphine, codeine, 6-acetylmorphine (6AM), heroin, 6-acetylcodeine (6AC), nicotine, cotinine, and trans-3′-hydroxycotinine (OH-cotinine) by liquid chromatography tandem mass spectrometry in oral fluid (OF) was developed and extensively validated. Acetonitrile (800 μL) and OF (250 μL) were added to a 96-well Isolute-PPT+protein precipitation plate. Reverse-phase separation was achieved in 16 min and quantification was performed by multiple reaction monitoring. The assay was linear from 0.5 or 1 to 500 μg/L. Intraday, interday, and total imprecision were less than 13% (n?=?20), analytical recovery was 92–114% (n?=?20), extraction efficiencies were more than 77% (n?=?5), and process efficiencies were more than 45% (n?=?5). Although ion suppression was detected for EME, cocaine, morphine, 6AC, and heroin (less than 56%) and enhancement was detected for BE and nicotine (less than 316%), deuterated internal standards compensated for these effects. The method was sensitive (limit of detection 0.2–0.8 μg/L) and specific (no interferences) except that 3-hydroxy-4-methoxyamphetamine interfered with AEME. No carryover was detected, and all analytes were stable for 24 h at 22 °C, for 72 h at 4 °C, and after three freeze–thaw cycles, except cocaine, 6AC, and heroin (22–97% loss). The method was applied to 41 OF specimens collected throughout pregnancy with a Salivette® OF collection device from an opioid-dependent BUP-maintained pregnant woman. BUP ranged from 0 to 7,400 μg/L, NBUP from 0 to 71 μg/L, methadone from 0 to 3 μg/L, nicotine from 32 to 5,020 μg/L, cotinine from 125 to 508 μg/L, OH-cotinine from 11 to 51 μg/L, cocaine from 0 to 419 μg/L, BE from 0 to 351 μg/L, EME from 0 to 286 μg/L, AEME from 0 to 7 μg/L, morphine from 0 to 22 μg/L, codeine from 0 to 1 μg/L, 6AM from 0 to 4 μg/L, and heroin from 0 to 2 μg/L. All specimens tested negative for EDDP and 6AC. This method permits a fast and simultaneous quantification of 16 drugs and metabolites in OF, with good selectivity and sensitivity.     

毛细管电泳-质谱联用法测定清肺抑火片中的四种有效成分

建立了毛细管电泳-电喷雾电离质谱联用法同时测定清肺抑火片中苦参碱、药根碱、氧化苦参碱、栀子苷4种药效成分含量的分析方法.采用未涂层石英毛细管,以35 mmol/L乙酸铵(含20％乙腈,pH=6.5)为缓冲溶液、60％异丙醇(含3.3 mmol/L乙酸)为鞘液,分离电压为28 kV时,各组分在11 min内达到基线分离.苦参碱、药根碱、氧化苦参碱、栀子苷的线性范围分别为0.030～150 mg/L、0.060～20 mg/L、0.060～80 mg/L、0.60～300mg/L,检出限分别为0.010、0.020、0.020、0.20mg/L.样品的加标回收率在91.0％～107％之间,相对标准偏差均小于4.9％.方法简便、快速、灵敏度高,已成功用于清肺抑火片中的苦参碱、药根碱、氧化苦参碱、栀子苷含量的同时测定.     

High speed hydrophilic interaction liquid chromatographic method for simultaneous determination of selected pharmaceuticals in wastewater using a cyano-bonded silica column

In this work, a fast analytical method based on hydrophilic interaction liquid chromatographic-Ultraviolet detection (HILIC-UV) using a short narrow bore cyano-bonded silica column packed with fully porous sub-2?µm particles has been developed for simultaneous determination of eight pharmaceuticals in wastewater. The method involved pre-concentration and clean-up by solid phase extraction using Oasis HLB extraction cartridges. The analytes were separated using a mobile phase consisted of acetonitrile and 5?mM ammonium acetate buffer (95:5?v/v) with a flow rate of 0.6?mL/min. The chromatographic separation was optimized in order to achieve short analysis time and good resolution for all analytes in a single run. Each analyte was detected at its maximum wavelength for higher sensitivity. All analytes could be separated in 5.7?min with resolution ≥2.7. The optimized method was validated based on linearity, precision, detection and quantification limits, selectivity and accuracy. The detection limits of the studied pharmaceuticals ranged from 0.6 to 3?µg/L, while limits of quantification were in the range from 2 to 10?µg/L with UV detection. The developed method is fast, reliable, cost-effective and could be used for the analysis of the studied analytes in other matrices such as food, pharmaceutical preparations and biological fluids.     

A CE‐LIF method based on long wavelength fluorescence labeling for the analysis of thiols in human urine

A rapid and robust CE method using a long wavelength fluorescent reagent 1,7‐dimethyl‐3,5‐distyryl‐8‐phenyl‐(2‐maleimide)difluoroboradiaza‐s‐indacene as the labeling reagent has been developed for the simultaneous determination of thiols, including glutathione, cysteine, homocysteine, N‐acetylcysteine, cysteinylglycine, and penicillamine. The derivatization reaction was carried out in 14 mmol/L pH 8.5 borate buffer at 30°C for 6 min and the labeled thiols derivatives were separated with the running buffer containing 30 mmol/L pH 7.4 phosphate, 30% v/v acetonitrile and 8 mmol/L SDS within 12 min. Detection limits ranged from 0.4 to 2.4 nmol/L. To demonstrate the capability of this method, it was applied to the analysis of thiols in human urine with recoveries of 92.4–105.6%. The derivatization reaction was much faster at milder conditions, and the analysis was rapider. Moreover, with excitation wavelength at long wavelength region, background interference from samples was reduced effectively. The present method seems to be a potential choice for quantifying thiols in human urine.     

A new method for screening and determination of diuretics by on-line CE-ESI-MS

A rapid, high-resolution and effective new method for analyzing 12 diuretics by CE-ESI-MS was established in this paper. Ten diuretics (except two neutral compounds) could be fast separated by CE with a DAD at 214 nm with a 20 kV voltage within 6 min, using a 50 microm id and 48.5 cm effective length uncoated fused-silica capillary in a 40 mM ammonium formate buffer (pH 9.40). CE was coupled to the mass spectrometer applying an orthogonal electrospray interface with a triple-tube sheath liquid arrangement. The sheath liquid was composed of isopropanol-water (1:1 v/v) containing 30 mM acetic acid with a flow rate of 4 microL/min. Mass spectrum was employed in the positive mode and both full scan mode and SIM scan mode were utilized. All 12 diuretics could be detected and confirmed by MS in a single analysis. Under optimized conditions, LODs for the 12 diuretics were in the range of 0.13-2.7 micromol/L at an S/N of 3, and the correlation coefficients R(2 )were between 0.9921 and 0.9978. The RDSs (n = 5) of the method was 0.24-0.94 % for migration times and 1.6-8.8 % for peak areas. The recoveries of spiked samples of 12 diuretics were between 72.4% and 118%. The real urine samples were injected directly for analysis, with only simple filtration through a 0.22 microm membrane filter in order to remove solid particles, which may cause capillary blockage. Based on the migration times and characteristic ions, the diuretics in urine samples were detected successfully. This CE-ESI-MS method for analyzing diuretics will hopefully be applied to doping control.     

MECC分离并测定高粱及玉米中7种农药类环境激素

用毛细管胶束电动色谱成功分离并测定了高粱和玉米中7种农药类环境激素(多菌灵、西玛津、莠去津、毒死蜱、溴氰菊酯、乙草胺和氯氰菊酯)的残留量。研究了电泳缓冲液及表面活性剂等因素的影响,在最佳分离条件(pH 9.0、20 mmol/L磷酸氢二钠+50 mmol/L SDS+5%乙腈为缓冲溶液,分离电压20 kV,检测波长222 nm,实验温度25℃,压力法进样,30 mbar×10 s)下,7种农药在28 min内得到基线分离,质量浓度与其峰面积在5.0～150μg.L-1范围内呈良好线性,检出限为0.6～3.0μg/L,回收率为97%～108%,相对标准偏差为2.2%～4.7%。该方法具有操作简单、快速方便及自动化程度高、重现性好等优点。     

水产品中甲基汞测定的液相色谱-原子荧光光谱联用方法的改进

对本实验室前期建立的食品中甲基汞的液相色谱-原子荧光光谱联用测定方法进行了改进。采用无毒的半胱氨酸代替有毒试剂巯基乙醇作为流动相中的配位剂,流动相组成为5%(v/v)乙腈-1 g/L半胱氨酸-50 mmol/L乙酸铵水溶液,使汞化合物分离时间缩短至8 min。在优化条件下,甲基汞标准曲线的线性范围为1～50 μg/L,检出限(S/N=3)为0.3 μg/L。采用超声波辅助5 mol/L HCl提取样品中的甲基汞,提取液经C18固相萃取小柱净化后进样。鱼、虾、贝等不同种类水产动物样品以及水产类膳食样品的甲基汞加标回收率为89%～112%。对标准参考物质NIST1566b、BCR464和GBW10029以及英国食品分析水平评估计划(Food Analysis Performance Assessment Scheme, FAPAS)的罐装鱼肉样品(样品编号07115)的测定结果与参考物定值相符,验证了该方法的可靠性与准确性。本方法可满足食品中甲基汞检测的需要。     

Application of liquid chromatography to the simultaneous determination of acetylsalicylic acid, caffeine, codeine, paracetamol, pyridoxine, and thiamine in pharmaceutical preparations.

This paper describes a rapid reversed-phase liquid chromatographic method, with UV detection, for the simultaneous determination of acetylsalicylic acid, caffeine, codeine, paracetamol, pyridoxine, and thiamine in pharmaceutical preparations. A reversed-phase C18 Nucleosil column is used. The mobile phase consists of 2 successive eluants: water (5 min) and acetonitrile-water (75 + 25, v/v; 9 min), both adjusted to pH 2.1 with phosphoric acid. Before determination acetylsalicylic acid is completely converted to salicylic acid by alkaline hydrolysis. Salicylic acid, caffeine, paracetamol, pyridoxine, and thiamine are all detected at 285 nm, whereas codeine is detected at 240 nm. Calibration curves were linear for salicylic acid, caffeine, paracetamol, and pyridoxine in the range of 50-500 mg/L, and for codeine and thiamine in the range of 50-1000 mg/L. The method was applied to the analysis of 13 fortified commercial pharmaceutical preparations. Recoveries ranged from 92.6 to 105.5%, with relative standard deviations of 1.1-5.8%.     

Determination of carbohydrates in tobacco by pressurized liquid extraction combined with a novel ultrasound-assisted dispersive liquid–liquid microextraction method

A novel derivatization-ultrasonic assisted-dispersive liquid–liquid microextraction (UA-DLLME) method for the simultaneous determination of 11 main carbohydrates in tobacco has been developed. The combined method involves pressurized liquid extraction (PLE), derivatization, and UA-DLLME, followed by the analysis of the main carbohydrates with a gas chromatography-flame ionization detector (GC-FID). First, the PLE conditions were optimized using a univariate approach. Then, the derivatization methods were properly compared and optimized. The aldononitrile acetate method combined with the O-methoxyoxime-trimethylsilyl method was used for derivatization. Finally, the critical variables affecting the UA-DLLME extraction efficiency were searched using fractional factorial design (FFD) and further optimized using Doehlert design (DD) of the response surface methodology. The optimum conditions were found to be 44 μL for CHCl₃, 2.3 mL for H₂O, 11% w/v for NaCl, 5 min for the extraction time and 5 min for the centrifugation time. Under the optimized experimental conditions, the detection limit of the method (LODs) and linear correlation coefficient were found to be in the range of 0.06–0.90 μg mL⁻¹ and 0.9987–0.9999. The proposed method was successfully employed to analyze three flue-cured tobacco cultivars, among which the main carbohydrate concentrations were found to be very different.