脂肪胺荧光衍生物的高效液相色谱分离及质谱鉴定

采用新型荧光衍生试剂2-(9-吖啶酮)-乙酸(AAA)进行柱前衍生并经荧光检测对脂肪胺进行了高效液相色谱(HPLC)分离和在线质谱定性.衍生物荧光激发和发射波长为λex=404nm,λem=440nm.30℃下在乙腈溶剂中用N-乙基-N′-[(3-二甲氨基)丙基]碳二亚胺盐酸盐(EDC)做催化剂,衍生反应20min后获得稳定的荧光产物.在HypersilBDSC18(4.6mm×100mm,5μm)色谱柱上,采用梯度洗脱对12种脂肪胺衍生物进行了优化分离.采用大气压化学电离源(APCISource)正离子模式进行在线柱后质谱定性,实现了各种脂肪胺衍生物的快速、准确测定.该方法具有良好的重现性,多数脂肪胺的线性回归系数大于0.9996,检测限为12.09~25.52fmol.     

脂肪胺的高效液相色谱分离及质谱鉴定

 采用新型荧光试剂1,2-苯并-3,4-二氢咔唑-9-乙酸(BCAA)为柱前衍生化试剂,在Hypersil BDS-C18色谱柱上,通过梯度洗脱对12种游离脂肪胺进行了分离和在线质谱定性。以乙腈为溶剂,1-乙基-3-(3-二甲氨基丙基)环己碳二亚胺(EDAC)为缩合剂,在50 ℃条件下衍生反应15 min后获得稳定的荧光产物。激发波长和发射波长分别为333 nm和390 nm。采用大气压化学电离源(APCI)的正离子模式,实现了土壤和污水中脂肪胺的定性及其含量的测定。脂肪胺的线性相关系数大于0.9993,检测限为12~28 fmol。     

污水中脂肪胺荧光衍生物的高效液相色谱分离及质谱鉴定

利用新型荧光试剂1,2-苯并-3,4-二氢咔唑-9-异丙基氯甲酸酯(BCIC)作为柱前衍生化试剂,在乙腈中,以硼酸钠缓冲液(pH 9)为催化剂,40 ℃下衍生反应10 min后获得稳定的荧光产物.在Eclipse XDB-C8色谱柱上,通过梯度洗脱对12种游离脂肪胺进行了分离和在线质谱定性.激发和发射波长分别为λex=333 nm,λem=390 nm.采用大气压化学电离源(APCI)正离子模式,实现了污水中脂肪胺的定性及相应含量测定.脂肪胺的线性回归系数大于0.9996,检出限在10.57～37.83 fmol.     

柱前衍生-高效液相色谱分离测定及质谱鉴定脂肪胺

采用新型荧光衍生试剂2-(2-苯基-1-氢-菲[9,10-d]咪唑)-乙酸(PPIA)进行柱前衍生,经荧光检测实现了脂肪胺的高效液相色谱(HPLC)分离测定及柱后质谱鉴定。60℃下在乙腈溶剂中用N-乙基-N′-[(3-二甲氨基)丙基]碳二亚胺盐酸盐(EDC)做缩合剂,衍生反应15min可获得稳定的荧光产物。脂肪胺衍生物荧光检测波长为380nm(激发波长为260nm)。在EclipseXDB-C8色谱柱上,采用梯度洗脱对12种脂肪胺衍生物进行了优化分离,测定了造纸厂废水、大鼠端脑和酸奶中脂肪胺的含量。经柱后在线质谱大气压化学电离源(APCISource)正离子模式实现了各种脂肪胺衍生物的质谱鉴定,借助对活性中间体的质谱解析确定了衍生反应的反应机理。该方法具有良好的重现性和回收率,多数脂肪胺的线性回归系数大于0.9996;检出限为3.1~18fmol(S/N=3∶1)。     

新型荧光试剂用于土壤中脂肪胺的高效液相色谱-质谱测定

利用新型荧光试剂2-(2-(10-蒽基)-苯并咪唑)-乙酸(ABIA)为柱前衍生化试剂,在Akasil-C18色谱柱上,通过梯度洗脱对12种游离脂肪胺进行了分离和在线质谱定性。以乙腈为溶剂,N-乙基-N′-[(3-二甲氨基)丙基]碳二亚胺盐酸盐(EDC)为缩合剂,在50℃条件下衍生反应20 min后获得稳定的荧光产物。激发波长和发射波长分别为260 nm和430 nm,采用大气压化学电离源(APCI)的正离子模式,实现了土壤中脂肪胺的定性及其含量的测定。脂肪胺的线性相关系数大于0.9990,检出限为11.72~25.63 fmol。     

荧光衍生-高效液相色谱分离及质谱鉴定芳香胺

含氮杂环光致发光分子经柱前衍生化,采用荧光检测及柱后在线质谱鉴定对废水中胺类化合物进行了测定.衍生物荧光激发和发射波长分别为λex=275 nm,λem=380 nm.在pH 8.0的硼酸钠缓冲液中,于40 ℃下衍生反应40 min后获得稳定的荧光产物.在Hypersil BDS C18柱上,采用梯度洗脱对5种芳香胺衍生物进行了优化分离.采用大气压化学电离源(APCI Source)正离子模式进行在线的柱后质谱定性,实现了5种芳香胺衍生物的快速、准确测定.方法具有良好的重现性,线性回归系数大于0.9998,检出限在71～81 fmol水平.     

高效液相色谱荧光测定及质谱鉴定土壤和苔藓中的脂肪酸

以吖啶酮-9-乙基对甲苯磺酸酯(AETS)作荧光衍生化试剂,建立了灵敏、简单的游离脂肪酸反相高效液相色谱测定方法。在Ec lipse XDB-C8色谱柱上,实现了19种游离脂肪酸(FFA)衍生物的完全基线分离。选取AETS摩尔数为脂肪酸的6倍,以DMF作溶剂,在85℃条件下以K2CO3作催化剂可获得稳定的荧光产物,条件温和、衍生产率高。利用柱后在线串联质谱以APC I大气压化学电离源正离子模式实现了各组分的质谱定性。对土壤和3种苔藓植物中(树藓、狭叶绢藓、曲尾藓)FFA组分定量结果表明,苔藓植物从土壤中富集了大量的游离脂肪酸。荧光检测的激发和发射波长分别为404 nm和440 nm。绝大多数脂肪酸的线性相关系数大于0.9996,检出限为12.3~43.7 fmol。本方法具有良好的重现性,用于实际样品测定,结果满意。     

新型荧光衍生试剂用于脂肪胺的反相高效液相色谱-荧光检测分析

利用新型荧光试剂4-(1H-菲并[9,10-d]咪唑-2-)苯甲酸(PIBA)进行柱前衍生并经荧光检测对脂肪胺进行了高效液相色谱(HPLC)分离和在线质谱定性。激发和发射波长分别为ex=261nm,em=443nm。80℃下在吡啶溶剂中用N-乙基-N’-[(3-二甲氨基)丙基]碳二亚胺盐酸盐(EDC)做催化剂,衍生反应10min后获得稳定的荧光产物。在EclipseXDB-C8色谱柱(4.6150mm,5mm)上,梯度洗脱对12种游离脂肪胺衍生物进行了优化分离。采用大气压化学电离源(APCI)正离子模式,实现了各种脂肪胺衍生物的测定。多数脂肪胺的线性回归系数大于0.9999,检测限为10.5～53.4fmol。     

高效液相色谱-荧光光度测定及质谱鉴定酒中脂肪醛

合成了一种新的荧光试剂[1,2-苯并-3,4-二氢咔唑-9-乙基肼基甲酸酯(以下简写作BCEC)]并在酒中脂肪醛高效液相色谱(HPLC)分离及测定中作为柱前衍生试剂.衍生反应在乙腈溶剂中,用三氯乙酸作催化剂并在60 ℃条件下进行,并能在10 min后获得稳定的荧光产物.用Zorbax Eclipse XDB C8(4.6 mm×150 mm,5 μm)作色谱柱,并以不同浓度的乙腈-水溶液作流动相进行梯度淋洗.采用荧光检测,检测的激发波长为333 nm,发射波长为390 nm.采用在线质谱检测对10种脂肪醛进行定性鉴定,醛的物质的量在0.166 5～83.3 fmol之间与其色谱峰面积呈线性关系,相关系数均优于0.999 9 ,被测各醛的检出限(S/N=3)在3.75～16.65 fmol之间.     

2-(11H-苯[a]咔唑)-乙酸在脂肪胺的高效液相色谱分离及质谱鉴定中的应用

采用新型荧光衍生试剂2-(11H-苯[a]咔唑)-乙酸(BCA)进行柱前衍生并经荧光检测对脂肪胺进行高效液相色谱(HPLC)分离和质谱定性.衍生物荧光激发和发射波长为λex=285 nm,λem=384 nm.60 ℃下在乙腈溶剂中用N-乙基-N′-[(3-二甲氨基)丙基]碳二亚胺盐酸盐(EDC)作催化剂, 衍生反应15 min后获得稳定的荧光产物.在Hypersil BDS C18 (4.0 mm×200 mm, 10 μm) 色谱柱上, 采用梯度洗脱对12种脂肪胺衍生物进行了优化分离.采用大气压化学电离源(APCI Source)正离子模式进行质谱定性, 实现了各种脂肪胺衍生物的快速、准确测定.脂肪胺的线性回归系数不小于0.999 8 , 检出限(S/N=3)为5.73 ～31.3 fmol.     

柱前荧光衍生脂肪醇的高效液相色谱分离及质谱鉴定

利用新型荧光试剂1,2-苯并-3,4-二氢咔唑-9-乙酸（BCAA）作为柱前衍生化试剂,以乙腈为溶剂,1-乙基-3-（3-二甲氨基丙基）环己碳二亚胺（EDAC）作为缩合剂,4-二甲氨基吡啶（DMAP）为催化剂,55℃下衍生反应25 min后获得稳定的荧光产物。在Eclipse XDB-C8色谱柱上,通过梯度洗脱对12种游离脂肪醇进行了分离和在线质谱定性。激发和发射波长分别为λex=333 nm,λem=390nm。采用大气压化学电离源（APCI）正离子模式,对洗发水（含十二烷基硫酸钠）中游离脂肪醇进行定性及测定。脂肪醇的线性回归相关系数大于0.9997,检出限在9.40-25.32 fmol。     

A fast method for determining low-molecular-mass aliphatic carboxylic acids by high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry

A fast quantitative high-performance liquid chromatographic separation method with atmospheric pressure chemical ionization mass spectrometric detection (HPLC-APCI-MS) was developed for the determination of low-molecular-mass aliphatic mono- and dicarboxylic acids typically present in different industrial process waters. A mixture of glycolic, lactic, a-glucoisosaccharinic, oxalic, maleic, fumaric, succinic, malic, glutaric, methylsuccinic, and adipic acids was separated using an RP chromatographic system. Adipic acid was used as an internal standard to calculate correlation coefficients for the acids studied. The chromatographic analysis of these acids was primarily carried out by means of gradient elution with an aqueous formic acid solution (0.15%, pH 2.5) and methanol using a modified C18 stationary phase. Good acid separation could be obtained for all acids by optimizing the chromatographic conditions. The method provides a simple sample preparation and faster analysis time compared to the traditional gas chromatographic methods, thus enabling almost real-time monitoring of these acids. Finally, the method developed was applied to the analysis of a complex mixture of aliphatic hydroxy carboxylic acids, which are formed as alkaline degradation products of carbohydrates during wood delignification and are present in the cooking spent liquor (black liquor).     

Simultaneous characterization of isoflavonoids and astragalosides in two Astragalus species by high-performance liquid chromatography coupled with atmospheric pressure chemical ionization tandem mass spectrometry

A method was developed for the simultaneous identification of astragalosides (AGs) and isoflavonoids (IFs) in the roots of Astragalus membranaceus and Astragalus mongholicus by HPLC coupled with atmospheric pressure chemical ionization MS/MS (HPLC-APCI-MS/MS). Diagnostic fragment ions of AGs and different group of IFs were obtained with one AG and eight IF standards analyzed by CID-MS, which were adopted as characteristic MS/MS fingerprints for further identification of these compounds in the two Astragalus species by using HPLC-APCI-MS/MS. A total of 20 IFs and 10 AGs were identified or tentatively identified. Among them, six IFs were detected in A. membranaceus for the first time and five IFs were firstly identified in A. mongholicus. The results indicate that HPLC-APCI-MS/MS is a powerful tool for the simultaneous characterization of IFs and AGs in complex matrix.     

Determination of aliphatic amines in water by gas chromatography using headspace solvent microextraction

The possibility of applying headspace microextraction into a single drop for the determination of amines in aqueous solutions is demonstrated. A 1 μl drop of benzyl alcohol containing 2-butanone as an internal standard was suspended from the tip of a micro syringe needle over the headspace of stirred sample solutions for extraction. The drop was then injected directly into a GC. The total chromatographic determination was less than 10 min. Optimization of experimental conditions (sampling time, sampling temperature, stirring rate, ionic strength of the solution, concentration of reagents, time of extraction and organic drop volume) with respect to the extraction efficiency were investigated and the linear range and the precision were also examined. Calibration curves yielded good linearity and concentrations down to 2.5 ng ml⁻¹ were detectable with R.S.D. values ranging from 6.0 to 12.0%. Finally, the method was successfully applied to the extraction and determination of amines in tap and river water samples. This system represents an inexpensive, fast, simple and precise sample cleanup and preconcentration method for the determination of volatile organic compounds at trace levels.     

Determination of aliphatic amines by high-performance liquid chromatography-amperometric detection after derivatization with naphthalene-2,3-dicarboxaldehyde

A simple and sensitive liquid chromatographic method has been developed for the determination of low molecular weight aliphatic amines after their pre-column derivatization with naphthalene-2,3-dicarboxaldehyde (NDA). Derivatization conditions, including the NDA concentration, reaction pH and reaction time have been investigated for method optimization. The chromatographic separation of five amines was performed on ABZ PLUS column using mobile phase of methanol-water (80:20, v/v) at a flow rate of 0.2 mL min⁻¹. The detection was carried out with a 6 mm glassy carbon electrode at the applied potential of 0.7 V versus Ag/AgCl reference electrode. The detection limits were between 23.3 and 34.4 nmol L⁻¹ of amines with a sample injection volume of 2 μL. The present method was applied for the determination of aliphatic amines in lake water. The recovery ranged 52.2-127.9%. The RSD in analytes retention time was less than 0.3% and 2.4% for intra- and inter-day analyses, respectively. The RSD in peak area was below 5.8% for both intra-day and inter-day analyses. The total analysis was completed within 20 min.     

High performance liquid chromatography coupled with atmospheric pressure chemical ionization mass spectrometry for sensitive determination of bioactive amines in donkey milk

In the present study we report on the optimization and validation of a sensitive high performance liquid chromatography atmospheric pressure chemical ionization mass spectrometry (HPLC–APCI–MS) method for the determination of 8 bioactive amines (histamine, tyramine, tryptamine, 2-phenylethylamine, cadaverine, putrescine, spermidine and spermine) in donkey milk samples. The method involves donkey milk pre-treatment to remove proteins and pre-column dansylation of the amines. HPLC in reversed phase mode has been used for bioactive amines separation and the operating condition of the APCI–MS system proved to be powerful and very efficient for peak assignment. The separation was accomplished in a short time with an excellent resolution for all the amine peaks. Quantification was carried out by monitoring the characteristic [M+H]⁺ ion of each amine derivative. The method sensitivity, linearity and repeatability were assayed with satisfactory results. The detection limits of the analysed amines ranged from 0.5 μg L⁻¹ to 15 μg L⁻¹; the highest LOD was for spermine. Also remarkably good recovery values were obtained; at the lowest spiking level (1 μg L⁻¹) the percent mean recoveries ranged from 77.7 to 109.7. Furthermore, as the investigations relate to a complex matrix as donkey milk, suitable studies on matrix effect were performed. Finally, the developed and validated method was applied to analyse 13 donkey milk samples. Among the identified bioactive amines, putrescine, spermine and spermidine proved to be the main amines in donkey milk. Their concentration levels in the present study were lower than the values determined in mature human, cow and sow milk.     

液相色谱–大气压化学电离串联质谱法测定油脂中9种胆固醇氧化物

建立一种准确测定油脂中9种胆固醇氧化物含量的同位素稀释液相色谱-串联质谱测定方法。向油脂样品中加入无水乙醇和60%氢氧化钾溶液,在室温下避光皂化22 h,使用乙醚-石油醚混合液(1∶1)作为提取溶液,采用液-液萃取方式提取目标物,再使用硅胶柱进行净化,采用5 mmol/L乙酸铵溶液与甲醇-乙腈混合液(1∶1)为双流动相梯度淋洗,使用Agilent ZORBAX SB-C18色谱柱分离,在大气压化学电离(APCI)正离子模式下,采用多反应监测模式内标法定量测定。在10~500μg/L范围内,9种胆固醇氧化物的质量浓度与色谱峰面积线性良好,相关系数(r2)均大于0.995,检出限均为0.5 mg/kg,在1,5 mg/kg两个添加水平时,加标回收率为71.2%~94.4%,相对标准偏差小于10%(n=6)。该方法满足动植物油脂中胆固醇氧化物含量测定需要。