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。     

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)。     

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.     

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.     

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.     

土壤和水中脂肪胺的高效液相色谱荧光测定及质谱鉴定

利用新型荧光试剂1,2-苯并-3,4-二氢咔唑-9-乙基氯甲酸酯(BCEOC)作为柱前衍生化试剂,在EclipseXDB-C8色谱柱上,梯度洗脱对12种游离脂肪胺进行了优化分离。40℃下在乙腈溶剂中以硼酸盐缓冲溶液作催化剂,衍生反应10min后获得稳定的荧光产物。激发和发射波长分别为λex=333nm,λem=390nm。采用大气压化学电离源(APCI)正离子模式,实现了土壤和造纸污水中脂肪胺的定性测定。采用荧光法进行分析物的定量测定。多数脂肪胺的线性回归系数大于0.999,检出限在18.65~38.82×10-15mol。方法具有稳定良好的重现性,对实际样品测定结果满意。     

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。     

柱前衍生-荧光检测-高效液相色谱-质谱法测定尿中游离多胺

用新型荧光试剂1,2-苯并-3,4-二氢咔唑-9-乙基氯甲酸酯作为柱前衍生化试剂,在乙腈中,以pH 9的硼酸钠缓冲溶液为催化剂,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5种多胺进行了分离和在线质谱定性.激发波长(λex)和发射波长(λem)分别为333 nm和390 nm.采用大气压化学电离源(APCI)正离子模式,实现了人尿中游离多胺的质谱定性及荧光定量测定.检出限(3S/N)在4.98～9.31 fmol.此方法应用于尿样中多胺的测定,并测得回收率在94%～98%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.     

柱前衍生-反相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

脂肪酸广泛分布于动植物体内,对于调节某些代谢作用和生理功能起着重要作用,其含量测定对进一步深入研究生理学和病理生理学以及对临床诊断具有重要意义[1-2]。某些动植物体内富含人体必需的多种脂肪酸,对其进行定量分析,对饮食营养和保健品开发同样具有重要指导意义。脂肪酸在     

A fluorous tag-bound fluorescence derivatization reagent, F-trap pyrene, for reagent peak-free HPLC analysis of aliphatic amines

We have developed a novel pre-column fluorescence derivatization reagent for amines, F-trap pyrene. This reagent comprises a fluorescent pyrene moiety, an amine-reactive Marshall linker, and a fluorophilic perfluoroalkyl group known as fluorous tag. When the reagent reacts with aliphatic amines and amino acids to give fluorescent derivatives, the fluorous tag in the reagent is eliminated simultaneously. Therefore, excess unreacted reagents in the derivatization reaction solution still have the fluorous tag and could be removed by fluorous solid-phase extraction selectively before high-performance liquid chromatography (HPLC) analysis. By using this reagent, 13 kinds of aliphatic amine (C₂–C₁₆) derivatives can be separated within 40 min by reversed-phase HPLC with gradient elution. In this chromatogram, unreacted reagents peak at around 28 min, greatly decrease after fluorous solid-phase extraction, and do not interfere with the quantification of each amine. The detection limits (S/N = 3) for examined aliphatic amines are 3.6–25 fmol per 20 μL injection. We have also applied this reagent successfully to the amino acid analysis.

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。     

高效液相色谱荧光测定及质谱鉴定分析血清中胆汁酸

利用新型荧光试剂1,2-苯并-3,4-二氢咔唑-9-乙基苯磺酸酯(BDEBS)作柱前衍生化试剂,在EclipseXDB-C8色谱柱上,采用梯度洗脱对10种胆汁酸(BAs)荧光衍生物进行了优化分离。95℃下在二甲基亚砜(DMSO)溶剂中以柠檬酸钾作催化剂,衍生反应35min后获得稳定的荧光产物,衍生反应完全。荧光激发和发射波长分别为λex=333nm,λem=390nm。采用大气压化学电离源(APCISource)正离子模式,实现了血清中胆汁酸的定性测定。分析物的定量测定采用荧光法进行。线性回归系数均在0.9996以上,检出限为22.36~44.57×10-15mol。     

HPLC of 9-fluorenylmethylchloroformate-polyamine derivatives with fluorescence detection

Summary There are a number of reagents available for fluorescent labelling of primary amines. These include dansyl chloride, o-phthalaldehyde, fluorescamine, and a new reagent, 9-fluorenylmethylchloroformate (FMOC), reported recently. This paper describes a reversed-phase HPLC procedure for the separation and fluorescence detection of polyamines following pre-column derivatization with FMOC. The polyamines studied by this method include putrescine, cadaverine, spermidine, and spermine. Experiments were carried out to determine maximum fluorescence excitation and emission wavelengths, optimum reaction pH, linear ranges, and minimum detection limits for each of the polyamines. The HPLC method includes a gradient program which provides complete separation from serum hydrolysate components and specificity for the four polyamines with detection limits ranging from 2 to 9 pg. This procedure was applied to hydrolyzed serum samples.     

Interaction between naphthyl-indolyl-ethenes and aliphatic amines: Effects on fluorescence and trans→cis photoisomerization

The fluorescence of two trans naphthyl-indolyl-ethenes in solvents of different polarity and H-bonding ability is quenched by aliphatic amines. The H-bonded complexes formed both in the ground- and in the first excited singlet state are responsible for the quenching in non-hydroxylated media. In methanol the interaction, which occurs only in the excited state, leads to the deprotonation of the indolyl-moiety. The consequences of the quenching on the trans→cis photoisomerization yield are reported.     

Simple HPLC determination of colistin in medicated feeds by pre-column derivatization and fluorescence detection

Summary A simple and sensitive HPLC method was developed for the determination of colistin antibiotic in feeds employing pre-column derivatization and fluorescence detection. Extraction of colistin in feeds was by sonication and shaking with 0.1 M HCl. Pre-column derivatization was with phthaldialdehyde and 2-mercaptoethanol (ME) in borate buffer (pH 10.5) to obtain a fluorescent derivative. Elution of the derivative onto an Ultracarb 5 μm ODS column was by using acetonitrile—ultrapure water (75∶25). Detection was by spectrofluorimetry at 340 nm (excitation wavelength) and 440 nm (emission wavelength). Total elution time was <20 min. The applicability of the validated method was tested by analyzing commercial medicated feeds without any interference from the matrix.