化学发光色谱柱后检测技术及其应用

化学发光(chemiluminescence,CL)检测的最大特点是设备简单、分析速度快及灵敏度高,是一种有效的微量和痕量分析手段,已在环境科学、临床检验、药学、工业分析等领域得到广泛的应用[1～3]。但是,化学发光分析法也存在着选择性差的问题,从而限制了这一方法的应用范围。高灵敏度的CL检测手段与高分辨的高效液相色谱(HPLC)、毛细管电泳(CE)和微流控芯片等分离技术相结合,成为一种理想的分离分析方法,得到人们的广泛重视。20世纪80年代的毛细管电泳和90年代初微流控芯片的出现、分析仪器的微型化和进样量少的要求向科研人员提出了要解决…     

鲁米诺化学发光体系在液相色谱检测中的应用

综述了鲁米诺化学体系在液相色谱检测中的应用，引用文献１９篇．     

一种新的液相色谱检测器——抑制化学发光反应检测器

化学发光分析法以其仪器设备简单、操作方便、分析快速、灵敏度高、线性浓度范围宽等显著优点,引起人们极大的兴趣。然而,由于化学发光反应固有的选择性差的缺点,使得这种高灵敏检测方法在分析应用中受到很大限制。如何将高灵敏度的化学发光检测法与高选择性的物理或化学手段相结合,这     

反相离子对液相色谱分离—柱后衍生化学发光检测痕量金…

报道了反相离子对液相色谱－柱后衍生化学发光检测方法分离测定痕量的钴（Ⅱ）、铁（Ⅱ）、铜（Ⅱ）、铁（Ⅲ）及铬（Ⅲ）。利用弱络合性的乳酸作为淋洗剂，加入适量的十二烷基磺酸钠作对离子试剂，研究了分离条件、发光条件及二者的匹配方式等因素对分离检测的影响。测定线性范围分别为（ｇ／ｍＬ）：Ｃｒ（Ⅲ）５．０×１０＾－９～１．０×１０＾－５；Ｆｅ（Ⅱ）２．０×１０＾－９～１．０×１０＾－５；Ｆｅ（Ⅲ）８．０×１０     

高效液相色谱-柱后化学发光法检测人体尿液中的卡托普利

在酸性条件下,Ce?氧化Ru(bipy)32+生成Ru(bipy)33+,同时氧化卡托普利生成二硫化物中间活性态([RS-SR]*),Ru(bipy)33+和二硫化物中间活性态之间相互反应产生强烈的化学发光。基于此,根据发光试剂Ru(bipy)32+水溶性好、试剂稳定等特点,将其加入到流动相中,通过高效液相色谱分离,建立了柱后化学发光快速灵敏检测卡托普利的方法。在以甲醇-0.01mol/L KH2PO4-1g/L Ru(bipy)32+(80∶20∶2,V/V)为流动相,流速为0.9mL/min,8.0×10-4 mol/L Ce?的优化实验条件下,方法的线性范围为2.0×10-7～1.0×10-4 mol/L(R2=0.9988),检出限为6.0×10-8 mol/L(S/N=3),并对1×10-5 mol/L卡托普利平行测定11次,相对标准偏差(RSD)为1.8%。将本方法用于人体尿液中卡托普利含量的测定,结果令人满意。结合化学发光光谱,对该体系发光机理进行了探讨。     

反相离子对液相色谱分离-柱后衍生化学发光检测痕量金属离子

报道了反相离子对液相色谱-柱后衍生化学发光检测方法分离测定痕量的钴(Ⅱ)、铁(Ⅱ)、铜(Ⅱ)、铁(Ⅲ)及铬(Ⅲ).利用弱络合性的乳酸作为淋洗剂,加入适量的十二烷基磷酸钠作对离子试剂,研究了分离条件、发光条件及二者的匹配方式等因素对分离检测的影响.测定线性范围分别为(g/mL):Cr(Ⅲ)5.0×10^-9～1.0×10^-5;Fe(Ⅱ)2.0×10^-9～1.0×10^-5;Fe(Ⅲ〕8.0×10^-9～1.0×10^-5;Cu(Ⅱ)8.0×10^-11～1.0×10^-5及Co(Ⅲ)1.0×10^-11～1.0×10^-5.检出限分别为:10pgCr(Ⅲ),0.4pgFe(Ⅱ),20pgFe(Ⅲ),1.0pgCu(Ⅱ)及0.01pgCo(Ⅱ).     

化学发光分析与高效液相色谱联用检测酪氨酸

在强碱性条件下,KMnO4氧化酪氨酸产生自由基,自由基氧化Luminol产生强化学发光,酪氨酸浓度与相对发光强度呈线性关系。选用C18反相色谱柱、CH3OH、KH2PO4混合液为流动相,酪氨酸能很好分离。基于此,建立CL-HPLC检测酪氨酸的新方法,线性范围为1.2×10-3～5.0×10-2g/L、线性相关系数为0.9994、RSD为2.37%、检测限为3.3×10-4g/L。方法已于检测实际样品氨基酸口服液,并与UV光度法,结果一致。本文探讨了酪氨酸增强KMnO4-Luminol化学发光机理。     

液相色谱化学发光检测法的新进展

本文评述了近年来液相色谱化学发光检测法的新进展，内容涉及各类化学发光发反应，生物发光反应和电致化学发光反应同色谱体系的偶合方式，仪器设计，多种无机，有机，生物大分子和生物活性物质的分析方法及其在环境，生物医学科学和生命科学中的应用和发展方向。引用文献１６８篇。     

液相色谱-化学发光检测法的最新进展

评述了近年来液相色谱－化学发光检测法的最新进展,参考文献从１９９２年到１９９５年。内容涉及各类化学发光反应同液相色谱体系的耦合方式,仪器设计,多种无机、有机、生物大分子、生物活性物质及药物的分析方法及其在环境、生物医学科学和生命科学、临床化学及药物化学中的应用和发展方向。     

Flow injection gas chromatography with sulfur chemiluminescence detection for the analysis of total sulfur in complex hydrocarbon matrixes

A fast and reliable analytical technique for the determination of total sulfur levels in complex hydrocarbon matrices is introduced. The method employed flow injection technique using a gas chromatograph as a sample introduction device and a gas phase dual‐plasma sulfur chemiluminescence detector for sulfur quantification. Using the technique described, total sulfur measurement in challenging hydrocarbon matrices can be achieved in less than 10 s with sample‐to‐sample time <2 min. The high degree of selectivity and sensitivity toward sulfur compounds of the detector offers the ability to measure low sulfur levels with a detection limit in the range of 20 ppb w/w S. The equimolar response characteristic of the detector allows the quantitation of unknown sulfur compounds and simplifies the calibration process. Response is linear over a concentration range of five orders of magnitude, with a high degree of repeatability. The detector's lack of response to hydrocarbons enables direct analysis without the need for time‐consuming sample preparation and chromatographic separation processes. This flow injection‐based sulfur chemiluminescence detection technique is ideal for fast analysis or trace sulfur analysis.     

Analysis of sulfur-containing compounds in crude oils by comprehensive two-dimensional gas chromatography with sulfur chemiluminescence detection

This paper reports an analytical method for separating, identifying, and quantifying sulfur-containing compounds in crude oil fraction (IBP-360 degrees C) samples based on comprehensive two-dimensional gas chromatography coupled with a sulfur chemiluminescence detector. Various sulfur-containing compounds and their groups were analyzed with one direct injection. 3620 peaks were detected including 1722 thiols/thioethers/ disulfides/1-ring thiophenes, 953 benzothiophenes, 704 dibenzothiophenes, and 241 benzonaphthothiophenes. The target sulfur compounds and their groups were identified based on the group separation feature and structured retention of comprehensive two-dimensional gas chromatography as well as standard substances. The quantitative analysis of major sulfur-containing compounds and total sulfur was based on the linear response of the sulfur chemiluminescence detector using the internal standard method. The sulfur contents of target sulfur compounds and their groups in 4 crude oil fractions were also determined. The recoveries for standard sulfur-containing compounds were in the range of 90-102%. The quantitative result of total sulfur in the Oman crude oil fraction sample was compared with those from ASTM D 4294 standard method (total S by X-ray fluorescence spectrometry), the relative deviation (RD%) was 4.2% and the precision of the method satisfactory.     

Comparison of combustion sources for sulfur chemiluminescence detection

Three different types of SCD combustion source have been evaluated for use in the chromatographic analysis of atmospheric sulfur compounds. The conventional FID source and the newer inverted burner source were found to be less sensitive and less stable than the flameless design. Overall, the flameless source was superior for use with HRGC-SCD.     

A post-column extraction system for the determination of tertiary amines by liquid chromatography with chemiluminescence detection

Summary The combination of an ion-pair extraction detection system with peroxyoxalate chemiluminescence detection has been investigated. Ion-pairs of protonated tertiary amines with a chemiluminescent counter ion are on-line post-column extracted to 1,2-dichloroethane containing bis(2,4,6-trichlorophenyl)oxalate (TCPO). Hydrogen peroxide is added to the organic phase by means of a solid-state perhydrit reactor. The influence of the base catalyst (imidazole) on the chemiluminescence reaction in apolar solvents was studied. Some sulfonated chemiluminophores were compared with respect to their chemiluminescence, ion-pairing and extraction properties, and 5-dimethylaminonaphthalene-1-sulfonate was found to be the most suitable reagent. The detection limit of the potential drug secoverine is in the sub-nanogram range.     

Determination of itopride hydrochloride by high-performance liquid chromatography with Ru(bpy)3 electrogenerated chemiluminescence detection

In this work, a stable electrogenerated chemiluminescence (ECL) detector was developed. The detector was prepared by packing cation-exchanged resin particles in a glass tube, followed by inserting Pt wires (working electrode) in this tube and sealing. The leakage of Ru(bpy)₃²⁺ can be compensated by adding a small amount of Ru(bpy)₃²⁺ into solution phase. Coupled with high-performance liquid chromatography separation, the detector has been used for determination of itopride hydrochloride in human serum. Under the optimal conditions, the ECL intensity has a linear relationship with the concentration of itopride hydrochloride in the range of 1.0 × 10⁻⁸ g mL⁻¹ to 1.0 × 10⁻⁶ g mL⁻¹ and the detection limit was 3 × 10⁻⁹ g mL⁻¹ (S/N = 3). The as-prepared ECL detector displayed good sensitivity and stability.     

Peroxyoxalate chemiluminescence detection with packed column supercritical fluid chromatography

Summary A detection scheme based upon peroxyoxalate chemiluminescence, which utilizes two post-column pumps and two stages of depressurization is investigated. The chemiluminescent detection limit for perylene is 23 times lower than determined by fluorescence, and is in the attomole range. This detection technique is investigated for packed column supercritical fluid chromatography (SFC). Due to the interface design used and the chemical band narrowing effects of chemiluminescence, an apparent increase in efficiency is observed. The interface design affords a wide range of pressures to be used for a separation. During pressure programming the column effluent changes flow rate. Because of a back-pressure regulator, the reaction and detection take place at nearly constant pressure. Therefore pressure gradient work is possible without concern for post-column reagent solubility (which is a concern for high-performance liquid chromatography). The effects of the expanded CO₂ from the SFC on the chemiluminescence signal and background are studied. The post-column detection is optimized for pH, photomultiplier voltage, concentrations and flow rates of the peroxide and oxalate ester.     

Direct electrogenerated chemiluminescence detection in high-performance liquid chromatography for determination of ofloxacin

Ofloxacin (OFLX) exhibited strong electrogenerated chemiluminescence (ECL) in NaNO₃ solution with a dual-electrode system when constant current was exerted. Based on this observation, a sensitive direct ECL method coupled with high-performance liquid chromatography (HPLC) separation was developed for determination of OFLX in human serum. Factors affected the ECL emission were investigated. Under the optimal conditions, the ECL intensity has a linear relationship with the concentration of OFLX in the range of 1.0 × 10⁻⁸ to 4.0 × 10⁻⁶ g mL⁻¹ and the detection limit was 4 × 10⁻⁹ g mL⁻¹ (S/N = 3). The proposed method was sensitive, simple and convenient to operate.     

Determination of vitamin K homologues by high-performance liquid chromatography with on-line photoreactor and peroxyoxalate chemiluminescence detection

A sensitive and highly selective high-performance liquid chromatography (HPLC) method was developed for the determination of vitamin K homologues including phylloquinone (PK), menaquinone-4 (MK-4) and menaquinone-7 (MK-7) in human plasma using post-column peroxyoxalate chemiluminescence (PO-CL) detection following on-line ultraviolet (UV) irradiation. The method was based on ultraviolet irradiation (254 nm, 15 W) of vitamin K to produce hydrogen peroxide and a fluorescent product at the same time, which can be determined with PO-CL detection. The separation of vitamin K by HPLC was accomplished isocratically on an ODS column within 35 min. The method involves the use of 2-methyl-3-pentadecyl-1,4-naphthoquinone as an internal standard. The detection limits (signal-to-noise ratio = 3) were 32, 38 and 85 fmol for PK, MK-4 and MK-7, respectively. The recoveries of PK, MK-4 and MK-7 were greater than 82% and the inter- and intra-assay R.S.D. values were 1.9-5.4%. The sensitivity and selectivity of this method were sufficient for clinical and nutritional applications.     

Determination of artemisinin in human serum by high-performance liquid chromatography with on-line UV irradiation and peroxyoxalate chemiluminescence detection

Artemisinin is an antimalarial drug containing an internal endoperoxide linkage in its structure. A simple, selective and sensitive high-performance liquid chromatography (HPLC)-peroxyoxalate chemiluminescence (PO-CL) method for the determination of artemisinin was developed. This method is based on the fact that endoperoxide in artemisinin structure can be converted to hydrogen peroxide (H(2)O(2)) under ultraviolet (UV) irradiation and the generated hydrogen peroxide can be measured using PO-CL detection. The HPLC-PO-CL system was optimized on a mobile phase, post column chemiluminescence reagent, UV source and irradiation time. In addition, the system was combined with simple liquid-liquid extraction using n-hexane that allowed selective and sensitive determination of artemisinin in serum. The limit of detection using 0.5 mL of blood was 0.062 micromol/L (17.5 ng/mL) at a signal-to-noise ratio of 3. Calibration curve obtained for artemisinin in human serum 4-80 micromol/L (1.1-22.6 microg/mL) showed a good linearity (r = 0.999).     

Detection of indomethacin by high-performance liquid chromatography with in situ electrogenerated Mn(III) chemiluminescence detection

The determination of indomethacin (INM) in pharmaceutical and biological samples by means of high-performance liquid chromatography (HPLC) with in situ electrogenerated Mn(III) chemiluminescence (CL) detection was proposed. The method was based on the direct CL reaction of INM and Mn(III), which was in situ electrogenerated by constant current electrolysis. The chromatographic separation was carried out on Nucleosil RP-C₁₈ column (250 mm × 4.6 mm; i.d., 5 μm; pore size, 100 Å) at 20 °C. The mobile phase consisted of methanol:water:acetic acid = 67:33:0.1 solution. At a flow rate of 1.0 mL min⁻¹, the total run time was 10 min. The effects of several parameters on the HPLC resolution and CL emission were studied systematically. Under the optimal conditions, a linear range from 0.01 to 10 μg mL⁻¹(R² = 0.9991), and a detection limit of 8 ng mL⁻¹ (signal-to-noise ratio = 3) for INM were achieved. The relative standard deviations (R.S.D.) for 0.1 μg mL⁻¹ INM were 2.2% within a day (n = 11) and 3.0% on 5 consecutive days (n = 6), respectively. The recovery of INM from urine samples was more than 92%. The applicability of the method for the analysis of pharmaceutical and biological samples was examined.