响应面法优化丹磺酰氯衍生生物胺的衍生条件

通过采用响应面法优化丹磺酰氯同8种生物胺反应的衍生温度、衍生时间和衍生剂用量,研究不同衍生因素之间的交互作用,以期为生物胺衍生条件的优化提供更为科学、直观的理论依据。研究结果表明:用10mg/m L的丹磺酰氯衍生20μg/m L的8种生物胺混标(工作)溶液,在衍生温度40. 06℃,衍生时间34. 70 min,衍生剂添加量3 m L时,生物胺的衍生产率最高,由模型得到生物胺总面积的预测值与验证试验的吻合率达到100. 05%,说明该模型能较好地预测丹磺酰氯衍生生物胺的衍生效果,具有较高参考价值。同时,通过响应面交互作用分析表明,在以8种生物胺总面积为研究对象时,衍生温度与衍生时间、衍生剂用量并没有显著的交互作用(P> 0. 05),衍生时间与衍生剂用量之间存在显著(P <0. 05)的交互作用。     

响应面法优化藜芦胺的提取工艺

为了优化藜芦胺的提取工艺,采用响应面法优化提取时间、提取温度和料液比,分析并建立数学模型.结果表明.提取时间、提取温度、料液比对藜芦胺的提取量均有极显著影响(p＜0.01);并确定最佳提取工艺参数为时间2h,提取温度75.0℃,料液比1∶9(g/mL)在此优化条件下,20.0g干燥藜芦根提取得到藜芦胺的量为98.23mg,与模型预测值得比较误差为2.66％.与模型预测值吻合,说明所建立的模型符合实际操作.     

生物胺丹磺酰氯衍生条件优化研究

研究了丹磺酰氯作为衍生试剂同5种生物胺反应的条件,优化了丹磺酰氯的浓度、反应体系的p H值、反应的温度和反应时间。研究结果表明:在腐胺、尸胺、组胺、酪胺和精胺浓度分别为8μg/m L的混合生物胺溶液时,5mg/m L丹磺酰氯丙酮溶液与生物胺底物溶液用量比为1∶1时衍生效果最佳,最佳反应p H值为10.5,最佳反应温度为60℃,最佳反应时间为20min。反应条件的初步确立对于进一步优化反应条件和准确定量生物胺含量具有重要意义。     

高效液相色谱法测定黄酒中生物胺的含量

本文建立了测定酿造酒(黄酒)中生物胺含量的高效液相色谱法,优化的测定条件为:样品经0.4mol/L高氯酸提取后用丹磺酰氯柱前衍生,流动相为乙腈和水,采用梯度洗脱,流速为0.8ml/min,二极管阵列检测器,检测波长为254nm。该方法检测限为:尸胺、组胺和亚精胺为0.05μg/ml,酪胺为0.1μg/ml,精胺为0.25μg/ml。线性范围为2.0～40.0μg/ml(R>0.99),回收率分别为尸胺96.6%、组胺101.94%、酪胺101.90%、亚精胺98.65%、精胺107.4%。首次采用该法测定了黄酒中的生物胺,结果表明黄酒中生物胺的种类及含量因酒的品种而异,5种生物胺平均总量为114.45μg/ml,变异范围为39.27～241.07μg/ml。     

白酒和黄酒中生物胺的高效液相色谱分析法

本研究建立了1种检测某白酒与黄酒中常见的9种生物胺(尸胺、组胺、酪胺、色胺、盐酸吡哆胺、精胺、亚精胺、腐胺和2-苯乙胺)的柱前衍生-高效液相色谱法,并对其实际运用可行性进行验证与讨论。以丹磺酰氯(Dns-Cl)作为衍生试剂,采用Thermo Hypersil GOLD C18色谱柱(250 mm×4.6 mm, 5μm)分离,以乙腈和含0.1%甲酸的0.01 mol/L乙酸铵水溶液作为流动相A与B,流速0.8 mL/min,紫外检测器波长254 nm。分析结果显示,9种生物胺在质量浓度0.5～50 mg/L范围,生物胺浓度与峰面积间线性相关性良好(R20.998)。选取1.0,5.0,10.0 mg/L 3个水平加标,得到加标回收率83.30%～114.7%,相对标准偏差均低于4.56%,9种生物胺检出限为0.07～0.22 mg/L。采用柱前衍生-高效液相色谱法分离、检测白酒与发酵酒——黄酒中的生物胺,实现了白酒与黄酒中生物胺含量的准确检测。     

高效液相色谱法同时测定酱油中的8种生物胺

本文建立了高效液相色谱法同时检测酱油中色胺、苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺的方法。采用5%三氯乙酸溶液为样品提取溶剂,提取液经丹璜酰氯柱前衍生30 min,最终HPLC进行定性和定量分析。本方法中8种生物胺的线性范围为1.0~50μg/mL,相关系数R2大于0.99,检出限(S/N=3)为0.075~0.3μg/mL。在添加水平为1.00和5.00μg/mL时,样品的平均回收率在83%~111%之间,相对标准偏差为0.43%~19.0%。本方法具有线性范围广,灵敏度和准确度高等优点,适用于酱油中生物胺的检测。采用本方法对市售10种酱油进行检测,总生物胺含量的范围为50.82~1898.17μg/mL,其中酪胺、腐胺和苯乙胺是酱油样品中含量最多的生物胺。     

响应面法优化野菊花多糖的水解工艺

在单因素实验基础上,应用响应面法对野菊花多糖的水解条件进行优化。以野菊花为研究对象,通过单因素实验确定水解时间、温度和三氟乙酸用量对野菊花多糖水解的影响,以气相色谱图中单糖的总峰面积为响应值,结合响应面试验,确定野菊花多糖的最佳水解条件。结果表明,野菊花多糖的最佳水解条件为:水解时间9 h,水解温度110℃,三氟乙酸用量为3 m L,在该水解条件下经过气相色谱测得的各个单糖峰面积及其总峰面积最大;模型预测值为4162.26,实际值为4151.35,与预测值相近,二者相对误差为0.26%。说明该模型优化所得的野菊花多糖的水解工艺合理,条件稳定可行。     

高效液相法测定市售腌制蔬菜商品中8种生物胺的含量

采用丹磺酰氯为衍生试剂,建立反相高效液相色谱法同时测定腌制蔬菜中色胺,β-苯乙胺,腐胺,尸胺,组胺,酪胺,亚精胺和精胺含量的方法。色谱条件:C18色谱柱分离,以水和乙腈为流动相梯度洗脱,流速0.8 mL/min,紫外检测器,检测波长:254 nm。在设定的试验条件下,8种生物胺在30 min内实现了良好的分离,待测生物胺峰面积同其相应浓度呈良好的线性相关性(R2>0.998)。仪器重复性良好(RSD<0.63%),方法重复性在可接收范围内(RSD<10%),8种生物胺的平均回收率在89.19%～103.02%之间。结果表明生物胺的HPLC检测方法,灵敏度高,精密度和重复性好,可快捷、准确地对腌制蔬菜中的生物胺进行检测。对市售腌制蔬菜样品中的生物胺含量进行了分析,检测出生物胺总量范围在39.38～628.82 mg/kg,属于目前我国安全标准范围之内。     

响应面法优化天麻发酵液发酵工艺

以Box-Behnken中心组合响应面试验设计法,优化天麻发酵液的最优发酵工艺。以鲜天麻为原料,在单因素试验的基础上,建立发酵时间、菌种添加量以及初始pH值对活菌数影响的二次多项回归模型,并验证模型的有效性。结果显示,天麻发酵液的最优发酵工艺条件为发酵时间42 h、菌种添加量1.26%、初始pH5.5,以此条件得到的天麻发酵液中活菌数为8.7 lg（CFU/mL）,与模型预测值基本吻合。实测结果与回归模型的相对误差为0.73%,说明采用响应面法优化得到的发酵条件可靠,有助于天麻发酵液衍生产品的深度开发。     

超高效液相色谱-串联质谱法同时测定蜂蜜中8种生物胺

建立了同时测定蜂蜜中色胺(TR)、2-苯乙胺(PHE)、腐胺(PUT)、尸胺(CAD)、组胺(HIS)、酪胺(TYR)、亚精胺(SPD)和精胺(SP)8种生物胺的超高效液相色谱串联质谱柱前衍生的分析方法。样品经5%三氯乙酸溶液提取后用丹磺酰氯柱前衍生,采用Agilent Poroshell 120 SB-C₁₈(50 mm×2.1 mm,2.7 μm)柱,以0.1%甲酸水溶液-0.1%甲酸乙腈溶液为流动相进行定性和定量分析。采用本方法对22种市售蜂蜜进行检测,结果显示8种生物胺在不同的蜂蜜中检出,所有蜂蜜样品中均检测到2-苯乙胺和腐胺,其中2-苯乙胺也是蜂蜜样品中平均含量最多的生物胺。本方法中8种生物胺的线性相关系数均大于0.99,检出限为0.01~0.51 μg/L。在添加水平为100、500、1000 μg/kg时,样品的平均回收率在76%~112%之间,日内精密度小于等于9.8%,日间精密度小于等于13.1%。该方法快速简单,灵敏准确,且回收率稳定,适用于蜂蜜中生物胺的检测。     

反相高效液相色谱-柱后衍生法分析检测鱿鱼中的生物胺

建立了水产品中多组分生物胺的反相高效液相色谱-柱后衍生-荧光检测方法。采用荧光试剂邻苯二甲醛（OPA）为柱后衍生化试剂,在Capcell Pak MG-C18色谱柱上,通过梯度洗脱使酪胺、腐胺、尸胺、组胺、胍丁胺、精胺和亚精胺等7种生物胺得到良好分离,在给定的浓度范围内,各组分生物胺呈现良好线性相关（R^2〉0.999）。在水产品中添加生物胺混合标准溶液,平均回收率为88.3%～110.1%,相对标准偏差RSD小于10%。结合水产品的感官鉴定、pH值和TVBN值测定等方法检测水产食品的新鲜程度,分析了鱿鱼在不同保藏温度、保藏时间下的生物胺种类及含量的变化。其中胍丁胺和尸胺在鱿鱼的保藏过程中发生最显著变化,可以作为其质量变化的参考指标。     

Biogenic Amine Content of Beers Consumed in Turkey and Influence of Storage Conditions on Biogenic Amine Formation

The biogenic amine content of beers available in Turkey, both domestic production and imported products, was investigated. A total of 17 domestic and 13 imported beers were evaluated in terms of 8 different biogenic amines (histamine, tyramine, spermine, spermidine, 2‐phenylethylamine, putrescine, tryptamine and cadaverine). HPLC methodology with pre‐column derivatization and photodiode array detection after derivatization with dansyl chloride was used for quantification. In addition, the evolution of important amines such as histamine, tyramine, putrescine and cadaverine were investigated during different storage conditions by response surface methodology. The results indicated that both storage temperature and storage time were important factors affecting biogenic amine content. Histamine and putrescine increased during storage, but then decreased after reaching a maximum level after six weeks. With the biogenic amines tyramine and cadaverine, the amounts increased during the entire storage period. At higher storage temperatures, the formation of biogenic amines increased.     

Validation of an HPLC Methodology for the Identification and Quantification of Biogenic Amines in Chicken Meat

This study validated a high performance liquid chromatography (HPLC) method to determine biogenic amines in chicken meat. For the identification of biogenic amines, an isocratic elution system coupled with a UV detector (254 nm) was used after a perchloric acid (5 %) extraction and benzoyl chloride derivatization of the samples. The standards of tyramine, putrescine, cadaverine, spermidine, and spermine were used for the following validation parameters: selectivity, linearity, accuracy, recovery, limit of detection and quantification, and robustness. Finally, chicken meat commercialized in two types of packaging was evaluated. The results showed an excellent selectivity and separation of all amines, r ²?>?0.99, relative standard deviation <5 %, recovery between 64 and 112 %, limits of detection and quantification between 0.03–1.25 and 0.15–5.00 μg?L^?1, respectively, and appropriate robustness for the proposed methodology. Moreover, both chicken meat commercial packages had similar values for all amines; only tyramine was significantly different (P?≤?0.05). The proposed method was suitable to detection and quantification of simultaneous five biogenic amines in chicken meat.     

两种柱前衍生RP-HPLC法测定酱腌菜中生物胺的比较

本研究分别采用4-氟-3-硝基三氟甲苯(FNBT)柱前衍生RP-HPLC紫外检测分析方法(简称FNBT衍生法)和丹磺酰氯柱前衍生RP-HPLC紫外检测分析方法(简称DNS-Cl衍生法)测定酱腌菜中生物胺含量,并比较这两种方法对酱腌菜中8种生物胺(组胺、色胺、β-苯乙胺、腐胺、酪胺、尸胺、亚精胺及精胺)检测的差异。结果表明,在浓度范围1~100mg/L内,两种方法对8种生物胺的检测呈现良好的线性相关性,相关系数R~2均大于0.999,FNBT衍生法的检出限(0.016~0.071mg/L)优于DNS-Cl衍生法(0.066~0.191 mg/L)。方法的精确度试验表明:FNBT衍生法的加标平均回收率(92.87%~100.18%)略优于DNS-Cl衍生法的加标平均回收率(83.52%~102.33%),两种方法的测量精密度RSD%均小于5%。DNS-Cl衍生法衍生试剂消耗大、操作繁琐,对生物胺检测的灵敏度低,而FNBT衍生法衍生试剂消耗少、操作简便、快速、灵敏度及准确性更高,适用于大批量食品中微量生物胺的快速、准确分析。用两种方法检测酱腌菜中8种生物胺的含量存在显著差异(p0.05)。     

Biogenic amine content of kefir: a fermented dairy product

Kefir is a fermented dairy product. Ten samples of kefir supplied from different manufacturers in Turkey were analyzed for the first time to determine biogenic amine contents using HPLC with benzoyl derivatization. Of the 10 biogenic amines under study, putrescine, cadaverine and spermidine were detected in all samples. Tyramine was detected in all kefir samples except one. Tyramine was the prevailing biogenic amine. Tyramine concentrations of kefir samples changed from non detectable values to 12.8 mg/l. Total biogenic amine contents of kefir samples were between 2.4 and 35.2 mg/l. Concentrations of biogenic amines were far below the allowable limits. pH values of kefir samples were in the range from 4.11 to 4.53; acidities were in the range from 0.652 to 1.047% (as lactic acid, w/v); total dry matters were from 8.88 to 12.00% (w/w); total free amino acid contents were from 0.0070 to 0.0206% (as leucine, w/v). No significant correlations were detected between biogenic amine concentrations and pHs and total dry matter contents. Significant correlations were obtained between biogenic amine concentrations and acidities and total free amino acid contents of kefir samples.     

Changes of yolk biogenic amine concentrations during storage of shell hen eggs

The RP-HPLC/UV method, using dabsyl derivatization, optimised for the determination of biogenic amines in egg yolk, was appropriate for quantification of putrescine, cadaverine, histamine, ethylamine, propylamine, ethanolamine, tyramine, tryptamine, spermine, spermidine, phenylethylamine. Detection limits ranged between 0.05 and 0.06 mg of biogenic amine/kg of egg yolk. Two experiments using, respectively, farm and avian eggs were conducted to evaluate yolk biogenic amine concentrations of fresh and stored eggs, and to explain the effect of temperature and time of storage in the levels of biogenic amines during egg shelf-life. Only five of the 11 biogenic amines under study were detected: putrescine, cadaverine, propylamine, ethylamine and ethanolamine. Storage time during shelf-life presented a significant effect on the levels of the five amines (p < 0.01). On the contrary, storage temperature did not presented a significant effect on the levels of the mentioned amines, p > 0.01. The significant reduction of biogenic amine concentration during the shelf-life justified the application of a multiple linear regression using stepwise method to estimate the storage time. The regression equation was applied with success to confirm the storage time of farm eggs and avian eggs that were stored at two different temperatures.     

Biogenic amine content of boza: A traditional cereal-based, fermented Turkish beverage

Boza is a fermented beverage made from millet, maize, wheat or rice. Biogenic amine contents of 10 boza samples from different manufacturers in Turkey were analysed for the first time, using HPLC after derivatisation with benzoyl chloride. Of the 11 biogenic amines under study, putrescine, spermidine and tyramine were detected in all boza samples. Tyramine was the prevailing biogenic amine. Tyramine concentrations of boza samples were between 13 and 65 mg/kg. Total biogenic amine contents of boza samples were between 25 and 69 mg/kg. Consequently, consumption of boza might represent a health risk for patients being treated with drugs containing monoamine oxidase inhibitor (MAOI). The pH values of boza samples were in the range from 3.16 to 4.02; total dry matters were from 15.3% to 31.1% (w/w); protein contents were from 0.50% to 0.99% (w/w). No significant correlations were detected between biogenic amine concentrations and pH, protein content and total dry matter content.     

不同贮藏温度下鲐鱼生物胺变化的研究

为探究不同温度和预处理对鲐鱼生物胺含量变化的影响,用高效液相色谱法分析了整鱼和去内脏鱼在不同贮藏温度下8种生物胺以及总生物胺含量的变化,结果发现,鲐鱼中主要的生物胺为组胺、腐胺、尸胺、酪胺。随着温度的升高,鲐鱼中总生物胺含量的增加迅速变快。在0、4、10、15、20、25和30 ℃贮藏1 d后,整鱼中总生物胺的积累量分别为40.34、93.44、107.95、73.39、119.99、4649.90、6446.43 mg/kg。在0、4、10、15、20、25和30 ℃贮藏条件下,整鱼中组胺的含量超过国家规定的高组胺鱼类安全限量(400 mg/kg)的时间分别为12、5、4 d和48、36、15、14~16 h,-18 ℃贮藏6个月,组胺积累量仅为13.45 mg/kg。另外,去内脏能减少生物胺的生成,在-18、0、4、10、15、20、25和30 ℃贮藏条件下,去内脏后总生物胺最终积累量减少程度分别为14.22%、39.79%、13.83%、29.06%、22.60%、13.56%、26.13%、21.26%。因此,低温冷冻和去内脏处理能够有效控制鲐鱼中生物胺的生成,防止腐败。本研究为有效控制鲐鱼生物胺的产生提供了参考依据。     

A Direct and Sensitive Analysis Method for Biogenic Amines in Dairy Products by Capillary Electrophoresis Coupled with Contactless Conductivity Detection

A capillary electrophoresis method coupled to contactless conductivity detection was developed for the quantitative determination of biogenic amines. Complete resolution of five biogenic amines, which are mostly found in dairy products, were performed in less than 6 min. Separation electrolyte composition was selected as 500 mM α-hydroxyisobutyric acid (HIBA) at pH 2.05. The detection limits of the method for biogenic amines were found between 0.041 and 0.098 mg/L. The precisions for the corrected peak areas were calculated as between 2.35 and 4.25% RSD. The applicability of the method was demonstrated by analyzing biogenic amines in the cheese, yoghurt, and kefir samples. The method does not require any pre-concentration and derivatization steps and can be applied as a simple, rapid, and precise method for the direct determination of biogenic amines in food samples.