离子色谱法同时测定水源水中的5种生物胺

建立了离子色谱(IC)同时测定水源水中5种生物胺(BA)(腐胺、尸胺、组胺、亚精胺和精胺)的方法。样品经0.45 μm水膜过滤后,用TSK-GEL SuperIC阳离子交换柱(150 mm×4.6 mm)分离,以甲磺酸水溶液为流动相梯度淋洗,流速为1.0 mL/min,非抑制电导检测,进样量为100 μL。实验结果表明,5种生物胺可以实现基线分离;在1.0～30.0 mg/L范围内,其峰面积与质量浓度之间的线性关系良好;保留时间的相对标准偏差(RSD)均不高于0.02%,峰面积的RSD小于2.08%;加标回收率为96.0%～107.0%。该法简便、快捷、准确,可以用于水源水中腐胺、尸胺、组胺、亚精胺和精胺5种生物胺的同时测定。     

沸水阻断多聚磷酸盐分解离子色谱法检测水产品中的多聚磷酸盐

多聚磷酸盐在水产品中的快速分解影响其准确测定.用沸水破坏使多聚磷酸盐分解的物质,并用三氯甲烷去除水产品中蛋白质等杂质,离子色谱准确测定多聚磷酸盐各组分的量.实验中采用淋洗液自动发生器产生的KOH溶液作为流动相,保护柱和分离柱采用Ionpac AG11-HC和Ionpac AS11-HC抑制电导检测,可以较好地分离所检测焦磷酸根和三偏磷酸根.该方法操作简单,检出限为10 mg/kg,当水产品中添加多聚磷酸盐标准物质浓度达到10～1000 mg/kg范围时,回收率在70%～110%之间,相对标准偏差小于12% .     

高效液相色谱法测定海水中游离态腐胺、亚精胺和精胺

使用高效液相色谱法测定海水中3种游离态多胺(腐胺、亚精胺和精胺).海水样品经 HClO4溶液酸化,丹磺酰氯衍生化,ODS色谱柱(150 mm × 4.6 mm i.d., 5 μm)分离,以0.1 mol/L乙酸铵和乙腈为流动相梯度淋洗,流速为1.0 mL/min,荧光检测(激发波长:340 nm;发射波长:515 nm).本实验中腐胺、亚精胺和精胺的检出限(S/N=3)分别为9.6×10-11, 2.8×10-10和1.0×10-10mol/L.在1×10-9～1×10-7mol/L范围内,3种多胺的浓度和荧光信号值均呈良好的线性关系(R>0.99).海水样品添加浓度在5×10-9, 1×10-8, 2.5×10-8和5×10-8 mol/L的回收率为84.9%～110.9%;衍生方法的相对标准偏差<3.6%,本方法中多胺衍生物不需要用有机溶剂萃取,衍生后可直接进样分析,避免了萃取过程中多胺的损失,极大地提高了分析速度.本方法具有选择性高,样品用量少,灵敏度好等特点,适合海水中游离态腐胺、亚精胺和精胺的痕量分析.     

离子交换色谱法对8种有机酸含量的同时测定

建立了淋洗液自动发生梯度淋洗的离子交换色谱法同时测定果汁中乳酸、乙酸、苹果酸、丙二酸、马来酸、草酸、柠檬酸、异柠檬酸8种有机酸的分析方法.样品经处理后用lonPac AS11-HC分离柱和lonPac AG11-HC型保护柱分离,以EG40自动淋洗液发生器生成的0.8～30 mmol/L KOH为淋洗液梯度洗脱,抑制型...     

高效液相色谱法同时测定水产品中10种生物胺的研究

高效液相色谱法同时测定水产品中色胺、2-苯乙胺、腐胺、尸胺、组胺、章鱼胺、5-羟色胺、酪胺、亚精胺和精胺。样品经高氯酸溶液提取，丹酰氯衍生化，C18柱色谱分离，以0．1mol／L醋酸铵和乙腈作流动相梯度淋洗，254nm紫外检测。方法检出限（SIN=3）：腐胺、亚精胺0．8μg／g；尸胺、组胺、酪胺、精胺1μg／g；章鱼胺、5-羟色胺2μg／g；2-苯乙胺3μg／g；色胺5μg／g。在0．2～10mg／L范围内，峰面积与质量浓度成良好的线性，相关系数r大于0．999。样品添加浓度在30、100和500μg／g的回收率为80％～111％，相对标准偏差为2．6％～15．8％。方法定量下限（LOQ）为30μg／g。本法简便、快速、灵敏、可靠。     

白酒中甜蜜素的无衍生离子色谱法检测

建立了免化学试剂离子色谱-抑制电导检测白酒中甜蜜素的方法。选用Dionex Ionpac AS17(250 mm×4 mm)分离柱,优化了电解水在线产生KOH淋洗液的梯度淋洗程序,样品无需衍生化,稀释后过0.22μm滤膜及OnGuardⅡRP离子色谱前处理小柱后直接进样检测,外标法定量。在0.05~20.0 mg/L范围内,甜蜜素的质量浓度与色谱峰面积呈良好的线性关系(r2=0.999 2),检出限为0.072 mg/L;峰面积和峰高的相对标准偏差(n=10)分别为1.6%、2.0%;加标回收率为85%~103%。该方法简便易行、误差因素少,无干扰、选择性好、灵敏度高、分析结果准确,适用于白酒中甜蜜素的检测。     

液相色谱-串联质谱法测定葡萄酒中7种生物胺含量

提出了液相色谱-串联质谱法测定葡萄酒中2-苯乙胺、色胺、尸胺、组胺、腐胺、精胺、亚精胺等7种生物胺含量的方法。样品经0.45μm滤膜过滤后,直接通过Zorbax Eclipse XDB C8色谱柱(4.6mm×150mm,5μm)进行分离,以不同体积5mmol·L-1乙酸铵-10mmol·L-1全氟己酸甲醇溶液(A)和5mmol·L-1乙酸铵-10mmol·L-1全氟己酸水溶液(B)的混合液为流动相梯度洗脱,串联质谱法进行测定。7种生物胺的质量浓度均在0.1～5.0mg·L-1范围内与其峰面积呈线性关系,检出限(3S/N)在0.005～0.02mg·L-1之间。以空白葡萄酒样品为基体进行加标回收试验,测得回收率在84.1%～97.5%之间;测定值的相对标准偏差(n=5)在6.3%～13%之间。     

稳定同位素稀释离子色谱-三重四极杆质谱法测定血浆和尿液中的氟乙酸

建立了离子色谱-三重四极杆质谱测定血浆和尿液样品中氟乙酸（MFA）的方法。血浆样品经高氯酸超声提取,尿液样品经高氯酸酸化,血浆和尿液提取液在pH 0.5~1.0条件下用叔丁基甲醚（MTBE）萃取,萃取液经氮吹浓缩后溶于0.1%（v/v）氨水溶液。以Ionpac AS 19型阴离子色谱柱为分析柱,在线自动产生的氢氧化钾作为淋洗液进行梯度分离,柱流出液经阴离子抑制器抑制后进入质谱系统。采用电喷雾电离源,在负离子、多离子监测（MRM）模式下检测,¹³C₂-氟乙酸稳定同位素内标法定量。血浆和尿液样品中氟乙酸的平均加标回收率为96.2%~120%,相对标准偏差为1.1%~13.1%（n=6）,方法的检出限（S/N=3）分别为0.03 μg/L和0.1 μg/L。该法简单、灵敏、准确,可用于生物样品中氟乙酸的检测。     

离子色谱-柱后衍生紫外检测法测定化妆品中碘酸盐与溴酸盐

建立了同时检测化妆品中溴酸盐和碘酸盐的离子色谱-柱后衍生紫外检测器定量分析方法。样品用水提取,经RP柱净化,采用IonPac AS11-HC色谱柱分离,以EG40自动淋洗液发生器生成的KOH为淋洗液梯度洗脱,溴化钾、亚硝酸钠为衍生试剂,二极管阵列检测器检测,外标法定量。结果表明:该方法在2.0～1 000 mg/kg范围内线性关系良好(r≥0.999 8);加标回收率为84%～99%;相对标准偏差小于2%。方法简单、快速、选择性强,可用于化妆品中碘酸盐和溴酸盐的测定。     

固相萃取-离子色谱法测定中药萝芙木中常见有机酸和阴离子

采用固相萃取-电导抑制离子色谱法测定了萝芙木中常见有机酸和无机阴离子。以Ionpac AS11为分离柱,Ionpac AG11为保护柱,萝芙木样品过IC SPE-C18前处理柱去除杂质和有机物,在确定的色谱条件下,柱温30℃,淋洗液为7.5 mmol/L Na OH溶液,流速为0.7 m L/min,电导检测器-离子色谱法直接测定样品,方法的相关系数在0.9987~0.9999之间,样品回收率在83.0%~100.8%之间,RSD5%。方法适用于中草药中常见有机酸和无机阴离子含量的测定。     

Determination of biogenic amines in fresh and processed meat by suppressed ion chromatography-mass spectrometry using a cation-exchange column

A new method for simultaneous determination of underivatized biogenic amines based on the separation by cation-exchange chromatography and suppressed conductivity coupled with mass spectrometry detection has been developed. The method has been applied to the analysis of cadaverine, putrescine, histamine, agmatine, phenethylamine and spermidine in processed meat products. The amines were extracted from muscle tissue with methanesulfonic acid without any additional derivative step or sample clean-up. Biogenic amines were separated by the IonPac CS17 column, a cation-exchange column used with gradient elution, and detection was done by suppressed conductivity and mass spectrometry. Tyramine was simultaneously analysed by using a spectrophotometer (275 nm) before the suppressed conductivity detection. Linearity of response was obtained in the range 0.25-25 microg mL(-1). The detection limits ranged from 23 microg L(-1) for putrescine to 155 microg L(-1) for spermidine (suppressed conductivity) and from 9 microg L(-1) for agmatine to 34 microg L(-1) for spermidine (MS). Average recoveries from meat samples ranged from 85 to 97% and coefficients of variation ranged from 4.5 to 9.7%. The analysis of biogenic amines in fresh and processed meats (dry-cured, cooked and fermented products) can be used as a quality marker of raw material and for studying the relationship between their changes and the fermentation process involved in dry sausage ripening.     

Determination of Biogenic Amines in Digesta by High Performance Liquid Chromatography with Precolumn Dansylation

A rapid high-performance liquid chromatographic method for the determination of nine biogenic amines in digesta sample from pig cecum and colon was developed using a simple direct derivatization with dansyl chloride. After precipitation with trichloroacetic acid and extraction by n-hexane, the amines were separated on a C₁₈ column using a water–acetonitrile gradient elution program. The calibration curve for each amine was linear over the range of 50 to 250 µmol/L, and the relative standard deviation for intra-assay precision was below 0.5%. The recovery ranged from 86.79% to 117.62% while the limit of detection was 0.1 µmol/L for the amines except for spermine with a value of 0.03 µmol/L. This procedure was successfully applied to identify and quantify biogenic amines in the hindgut digesta of the pig.     

Simultaneous determination of twelve biogenic amines in serum by high performance liquid chromatography

In this study, we established a method for simultaneously determining twelve biogenic amines in serum by using reversed phase high performance liquid chromatography (RP-HPLC). The biogenic amines were first extracted from human serum by perchloric acid solution and derivatized by dansyl chloride. An ODS column was selected as separation column at 40 °C. The mobile phase solutions were consisted of A, 0.1 mol/L ammonium acetate and B, acetonitrile. A gradient elution was carried out with a flow rate at 1.0 ml/ml. The results show that the detection limit for twelve biogenic amines ranged between 0.0621 and 0.628 μg/L. All the correlation coefficients were above 0.999. The linearity was over the range from 0.001 to 20 mg/L depending on individual biogenic amine. The intra-day and inter-day coefficients of variations were from 0.53% to 7.50%,and from 1.10% to 7.25% respectively. The average analytical recovery in serum was from 92.02% to 107.65%. Moreover, the serum concentrations of tryptamine, tyramine and histamine in healthy females were found lower than that in healthy males significantly. The method is sensitive, convenient, and reliable, and suitable for simultaneous analysis of multiple biogenic amines in the clinical diagnosis and drug discovery.     

Determination of trace biogenic amines with 1,3,5,7-tetramethyl-8-(N-hydroxysuccinimidyl butyric ester)-difluoroboradiaza-s-indacene derivatization using high-performance liquid chromatography and fluorescence detection

A reversed-phase high-performance liquid chromatographic method based on chemical derivatization with fluorescence detection has been developed for analyzing biogenic amines in food and environmental samples. A BODIPY-based fluorescent reagent, 1,3,5,7-tetramethyl-8-(N-hydroxysuccinimidyl butyric ester)-difluoroboradiaza-s-indacene (TMBB-Su), was employed for the derivatization of these biogenic amines at 20 °C for 20 min in pH 7.20 borate buffer after careful investigation of the derivatization conditions including reagent concentration, buffer solution, reaction temperature and reaction time. Separation of biogenic amines with gradient elution was conducted on a C8 column with methanol-tetrahydrofuran-water as mobile phase. The detection limits were obtained in the range from 0.1 to 0.2 nM (signal-to-noise=3). This procedure has been validated using practical samples. The study results demonstrated a potential of employing high-performance liquid chromatography (HPLC) with 1,3,5,7-tetramethyl-8-(N-hydroxysuccinimidyl butyric ester)-difluoroboradiaza-s-indacene labeling as a tool for quantitative analysis of biogenic amines involved in various matrices.     

在线固相萃取-毛细管高效液相色谱联用测定奶酪中的生物胺

采用双二元泵毛细管液相色谱,通过六通阀实现了样品的在线净化与分离定量的自动切换,建立了同时测定奶酪中的15种生物胺的在线固相萃取-毛细管高效液相色谱联用方法。通过优化毛细管高效液相色谱的分离条件,考察在线固相萃取流动相的组成、上样溶液pH值以及六通阀的切换时间对生物胺回收率的影响,确定最佳分析条件为：5%乙腈-水作为固萃柱(Zorbax SB-C18)的流动相,上样溶液pH=11,上样3 min后切换六通阀。采用内标法定量,15种生物胺标准曲线的线性范围为0.25~50.0 mg/L,检出限( LOD)为0.05~0.25 mg/L,定量限(LOQ)为0.15~0.80 mg/L。除了甲胺、乙胺、3-甲基丁胺和5-羟基色胺外,其余生物胺的不同添加水平(1,20和40 mg/kg)下的加标回收率为79.6%~118.7%;除3-甲基丁胺和5-羟基色胺外,其余生物胺的RSD在0.3%~14.9%之间,可用于奶酪中多种生物胺的快速检测。     

多壁碳纳米管净化-超高效液相色谱-质谱法测定水产品中头孢菌素残留量

建立了水产品可食部位中头孢哌酮、头孢哇肟、头孢洛宁、头孢唑啉、头孢匹林、头孢噻呋、头孢匹罗和头孢氨千8种头孢菌素的超高效液相色谱-质谱测定法.样品经乙睛-水溶液提取、多壁碳纳米管固相萃取净化后,以Acquity Xselect CSH C18柱为分离柱,用乙睛和0.1%甲酸溶液进行梯度洗脱,电喷雾正离子多反应模式监测.结果表明,8种头孢菌素均呈良好的线性关系(R2≥0.995),定量限(S/N=10)在2~10μg/kg之间;在阴性样品采取梯度加标,添加回收率为67.3%~94.2%,RSD为3.3%~14%.本方法检测成本低、准确度高、精密度好,能够满足水产品中头孢菌素检测的要求.     

反相硅胶整体柱离子对色谱法快速测定吡啶离子液体阳离子

建立了整体柱离子对色谱-紫外检测法梯度淋洗快速分离测定4种吡啶离子液体阳离子的方法。分离采用C18反相硅胶整体柱,以离子对试剂(用柠檬酸调节pH值)-乙腈为淋洗液,并采用多级梯度洗脱程序。实验考察了色谱柱、离子对试剂、乙腈浓度、色谱柱温度及流速对吡啶阳离子保留的影响,并讨论了其保留规律。咪唑阳离子的保留符合碳数规律。最佳色谱条件是:在流速3.0 mL/min,柱温30℃下,以1.0 mmol/L庚烷磺酸钠(pH 4.0)(A)+乙腈(B)为淋洗液进行梯度洗脱。淋洗梯度为0～2.0 min,10%B;2.0～2.5 min,10%～15%B;2.5～4.0 min,15%B;4.0～4.5 min,15%～20%B;4.5～10.0 min,20%B。在此条件下,4种吡啶阳离子可在7 min内基线分离。所测阳离子的检出限(S/N=3)为0.05～0.17 mg/L;峰面积的相对标准偏差(n=5)小于0.6%。将本方法用于实验室合成的离子液体样品和污水样品的分析,加标回收率在95.7%～99.0%之间。本方法准确、快速,具有较好的实用性。     

Determination of tocopherols,tocopherolquinones and tocopherolhydroquinones by gas chromatography-mass spectrometry and preseparation with lipophilic gel chromatography

A method has been developed for the simultaneous determination of a range of aromatic amines using cation-exchange chromatography performed on a standard ion chromatography column using d.c. amperometric detection. The analytes separated were 2,4- and 2,6-toluenediamine (2,4- and 2,6-TDA), aniline, o-toluidine, benzidine, p-chloroaniline, 4,4'-diaminodiphenyl (4,4'-DDP), m-nitroaniline and 1-naphthylamine. A Dionex CS12 column was used with gradient elution from an initial eluent of 5% CH3CN+35 mM H2SO4 to 27% CH3CN+35 mM H2SO4 (at 35 min). Detection limits in the range 2.6-22.6 microg/l were observed for all analytes except m-nitroaniline, for which the detection limit was 201 microg/l. Linear calibrations and good precision were observed and the method was applied to the determination of benzidine, p-chloroaniline and 1-naphthylamine in wastewater samples. Further, the separation was also used (after some modification of the eluent conditions) for the determination of 2,4- and 2,6-toluene diisocyanate (2,4- and 2,6-TDI) and 4,4'-methylenediphenyl diisocyanate (4,4'-MDI) after their hydrolysis to 2,4-TDA, 2,6-TDA and 4,4'-DDP. Detection limits for 2,6- and 2,4-TDI and 4,4'-MDI were 3.8, 8.2, and 11.2 microg/l, respectively. The method was applied to the determination of diisocyanates in air.     

Selective liquid-chromatographic determination of native fluorescent biogenic amines in human urine based on fluorous derivatization

A liquid chromatographic (LC) derivatization method for simple and selective determination of catecholamines and indoleamines in human urine has been developed. This method uses "fluorous interaction" in which perfluoroalkyl compounds show affinity with each other. The amino groups of native fluorescent analytes are precolumn derivatized with a non-fluorescent fluorous isocyanate, 2-(perfluorooctyl)ethyl isocyanate, and the fluorous-labeled analytes are retained in the fluorous LC column, whereas underivatized substances are not. Only the retained fluorous-fluorescent analytes are detected fluorometrically at appropriate retention times, and retained amines without fluorophores are not detected. In this study, 3,4-dihydroxyphenylalanine, dopamine, norepinephrine, epinephrine, and metanephrine were used as the representative of catecholamines. Tryptophan, 5-hydroxytryptophan, and 5-hydroxytryptamine were used as the representative indoleamines. This method was applied to determine eight biogenic amines in urine from healthy humans. The fluorous-labeled amines could be separated by fluorous LC column under conditions of isocratic elution within 35 min and simultaneously determined without interference from contaminants in biological samples. The detection limits for eight biogenic amines were 31-640 fmol on column. Calibration curves of them were linear over the range of at least 10-100 nmol/mL urine (r2 > 0.9989) with good repeatability.