离子色谱-脉冲安培检测法测定食品中的聚葡萄糖

目的建立食品中添加的聚葡萄糖(polydextrose, PDX)的离子色谱-脉冲安培检测方法。方法采用热水浸提聚葡萄糖,超滤离心去除高分子量的干扰物,滤液经酶解去除其他低聚糖的干扰,采用Hamilton RCX-30-250/4.6色谱分离柱;以含有A:150 mmol/L氢氧化钠和B:150 mmol/L氢氧化钠+500 mmol/L乙酸钠的淋洗液进行梯度淋洗,采用三电位波形,以同源标准物质建立标准曲线,流速1.2 ml/min,柱温32℃。结果本方法在30～200μg/g范围内,线性良好,r~2=0.9993。在检测固体样品时检出限为1.6699 mg/kg,在检测液体样品时检出限为0.9555 mg/kg,重复性测量相对标准偏差小于4.88%(n=6),不同食品基质聚葡萄糖的平均加标回收率为95.06%～110.11%。结论该方法灵敏度高,准确性好,结果稳定,适用于不同食品中添加的聚葡萄糖的测定。     

离子色谱-脉冲安培法测定饮料中的聚葡萄糖的含量

通过优化标准方法AOAC 2000.11和GB 5009.245-2016的样品前处理过程,简单、高效准确地分析饮料中添加的聚葡萄糖含量。方法 样品经离心超滤净化,去除基质干扰,采用色谱柱分离,离子色谱-脉冲安培检测器测定,外标法定量。结果 聚葡萄糖在0.20 ~ 2.00 g/L的浓度范围内呈良好线性关系 (R2 > 0.999)。方法检出限和定量限分别为0.01 和0.03 g/L,三个添加水平的平均加标回收率为90.0%~98.8%,相对标准偏差为0.63%~2.36%。同时考察了市场上10种不同品类的饮料,加标回收率为93.0%-105%。结论 本方法样品前处理简单、快速、广泛适用于饮料中添加的聚葡萄糖含量测定。     

离子色谱-脉冲安培检测器分析奶粉中低聚果糖

建立了离子色谱-脉冲安培检测分析奶粉中低聚果糖的方法。采用CarboPacPA200分离柱对分离条件进行了优化,确定柱温为30℃、淋洗液梯度洗脱,在10．0～50．0mg·L^-1范围内低聚果糖呈良好线性,回收率蔗果三糖（GF3）73．1％-82．6％、蔗果四糖（GF4）76．4％-82．9％、蔗果五糖（GF5）73．8％-77．6％,检出限分别为蔗果三糖10mg·kg^-1、蔗果四糖20mg·kg^-1、蔗果五糖24mg·kg～,精密度分别为蔗果三糖3．22％～4．61％、蔗果四糖4．60％-4．94％、蔗果五糖1．82％-3．26％。检测了乳粉中的低聚果糖．效果良好。     

抑制性电导检测-离子色谱法快速测定 水产品中的生物胺

目的建立抑制性电导检测-离子色谱法快速测定水产品中常见的几种生物胺。方法以甲基磺酸(MSA)为淋洗液通过梯度洗脱分离目标生物胺,采用Ionpac CS17(4 mm×250 mm)分析柱和电导检测器进行分析。进样体积为25μL,采用峰面积定量。通过比较不同的提取剂(TCA(三氯乙酸)、HCl O4、MSA)和提取方式(振荡、超声)的提取效果优化前处理方法。结果使用MSA超声提取效果较好,该方法测定生物胺的线性范围为0.02 mg~10.0 mg,检出限在0.5 mg/kg以下,加标回收率分别为85.2%~106.9%。结论该方法前处理简单快捷,结果灵敏、准确,适用于水产品中常见生物胺的快速检测。     

Dabsyl derivatisation as an alternative for dansylation in the detection of biogenic amines in fermented meat products by reversed phase high performance liquid chromatography

The commonly applied HPLC method to determine biogenic amines in dry fermented meat after dansylation was compared with an alternative dabsylation procedure. The use of dabsyl chloride at 70 °C resulted in a 25-min reduction of the derivatisation time, in comparison with the dansylation at 40 °C. Furthermore, the use of irritating ammonia to remove the excess of dansyl chloride can be avoided. Introduction of the SPE cleaning procedure on the C18 cartridge resulted in a reliable and sensitive method of biogenic amines determination in a complex protein–fat matrix, which is typical of dry fermented sausages.     

Optimisation of a chromatographic procedure for determining biogenic amine concentrations in meat and meat products employing a cation-exchange column with a post-column system

The aim of this study was to optimise and validate a chromatographic method for determining biogenic amine (BA) in meat and meat products separated by a cation-exchange column with a post-column system, using o-phthalaldehyde as a derivatising reagent. A perfect separation of nine BA (tyramine, histamine, β-phenylethylamine, putrescine, cadaverine, tryptamine, agmatine, spermidine and spermine) was obtained in 22 min. The conditions were: column Tª 40 °C and coil Tª 45 °C, pump flow rate 0.8 mL/min, pH phase A 6.33, B 5.63 and C 13.00. The method was adjusted linearly in a range of 0.10–12 mg/L with a correlation coefficient superior to 0.998. Detection and quantification limits were between 0.03–0.10 mg/L and 0.10–0.20 mg/L, respectively. Precision studies were satisfactory, with RSD less than 2% and meat extracts recovering over 98%. This method showed an appropriate, precise, fast and versatile procedure for determining nine BA simultaneously in different meat product matrices.     

离子色谱-脉冲安培法测定白酒中氰化物

目的建立离子色谱-脉冲安培法检测白酒中氰化物含量的方法。方法白酒样品稀释10倍后直接进样,以氢氧化钾溶液(12 mmol/L)进行等梯度淋洗,采用银工作电极,以Ag/AgCl复合参比电极模式,脉冲安培三点位波形方式进行检测。结果该方法在5～200μg/L浓度范围内线性关系良好,相关系数R~2为0.999 9。该方法对白酒类基质检出限为3.5μg/L,定量限为9.5μg/L。对不同类型白酒样品,在10、30和50μg/L三个加标浓度水平下回收率在91.7%～102.0%之间。日内相对标准偏差(RSD)在2.2%～4.8%之间,日间RSD在1.5%～1.9%之间。本方法与GB 5009.36—2016《食品安全国家标准食品氯化物的测定》中的气相色谱法进行比较,结果差异无统计学意义(P0.05)。结论该方法操作步骤简单快速,结果准确可靠,适用于实际白酒样品中氰化物的检测。     

In vitro synthesis of biogenic amines by Brochothrix thermosphacta isolates from meat and meat products and the influence of other microorganisms

Twenty Brochothrix thermosphacta strains tested for biogenic amines (BAs) production, formed histamine (6.6-16.2 mg/kg) and tyramine (18.7-35.4 mg/kg) but neither putrescine nor cadaverine. Six of the twenty strains were also investigated in respect of their influence on the synthesis of BAs by Escherichia coli, Pseudomonas sp., Proteus mirabilis and Lactobacillus sakei. In pure culture Escherichia coli produced all of the studied amines (histamine, tyramine, putrescine and cadaverine) with a total concentration of 167.7 mg/kg, P. mirabilis produced a total of 56.7 mg/kg histamine, tyramine and cadaverine, while Lactobacillus sakei and Pseudomonas sp. produced histamine and tyramine, totaling 37.9 and 35.2 mg/kg respectively. All B. thermosphacta promoted cadaverine production by Escherichia coli which increased by 12-68%, and some of them contributed to the appearance of this amine among the metabolites of Pseudomonas. The presence of B. thermosphacta decreased the potential ability of P. mirabilis to produce BAs.     

超高效液相色谱紫外-柱前衍生法测定肉制品中的9种生物胺

摘 要：目的 建立超高效液相色谱紫外-柱前衍生法测定肉制品中的9种生物胺。方法 实验选取西式火腿肠为样品,以三氯乙酸/高氯酸混合溶液对样品中生物胺进行提取,调节提取液于pH 10碱性环境下,使用1 mL的10 mg/mL丹磺酰氯溶液进行衍生化处理,60℃水浴衍生15 min,100 μL氢氧化钠中止衍生,乙酸乙酯进行最终萃取。使用超高效液相色谱仪紫外配二极管阵列检测器进行测定,采用流动相：1%乙酸乙腈-10 mmol/L乙酸铵溶液;波长：254 nm;柱温：35℃;进样量：3 μL;流速：0.3 mL/min;ACQUITY BEH C18(2.1 mm*100 mm, 1.7 μm)色谱柱进行分析。结果 在线性范围5~250mg/kg内9种生物胺的相关性均＞0.999,检出限范围为0.2~0.5 mg/kg,定量限范围为0.5~1.5 mg/kg,9种生物胺的日内精密度相对标准偏差RSD (relative standard deviation)范围为1.58%~ 5.21%,回收率均≥85%。结论 本研究所建方法具有良好的准确性、精密度、灵敏度,可为我国肉制品中生物胺的检测提供参考依据。     

Determination of chlorine ions in raw milk by pulsed amperometric detection in a flow injection system

Chloride ion concentration in milk was determined by pulsed amperometric detection in a flow injection system. Results showed that the Au electrode lost 3 electrons at 1.10 V and formed chloroaurate ions (AuCl₄^?) by combining with chloride ions, after which AuCl₄^? was partly reduced to Au at 0.6 V. Based on the electrochemical process, a triple waveform with detection potential of 1.15 V, detection time of 150 ms, oxidation potential of 1.4 V, oxidation time of 550 ms, reduction potential of 0 V, and reduction time of 400 ms was applied to the Au electrode for detecting chloride ion concentration in milk. The approach is rapid and automatic and features a detection limit of 0.005 g/L. The relative standard deviation obtained by 60 repetitive injections reached 1.48% at 2 g/L of NaCl. The method developed using the Au electrode without modification was used to analyze the chloride ion concentration in raw milk without preprocessing. The method showed good agreement with potentiometric titration.     

离子色谱法测定奶糖中的蔗糖、葡萄糖和乳糖

建立离子色谱法分离检测奶糖中蔗糖、葡萄糖和乳糖的分析方法。采用METROSEPCARB1（150mm×4.0mm）阴离子交换分离柱和脉冲安培检测器,对淋洗分离条件进行优化。确定柱温为32℃、淋洗液为37.0mmol/L NaOH溶液,淋洗液流速为1.0mL/min,总分析时间20min。在1.0～60.0mg/L范围内蔗糖、葡萄糖和乳糖分别呈良好线性,相关系数分别为0.9996、0.9998和0.9996,检出限分别为0.091、0.014和0.083mg/L,RSD为1.42%～5.17%,回收率为91.27%～101.31%。测定结果为大白兔奶糖中含葡萄糖1.56%、蔗糖21.40%、果糖3.75%;旺仔牛奶糖中含葡萄糖1.17%、蔗糖30.29%、果糖3.30%;金丝猴奶糖中含葡萄糖1.08%、蔗糖20.86%、果糖3.31%。用该方法检测了奶糖中的三种糖,效果良好,同时具有操作简单,结果准确,快速有效等优点,可为奶糖的检测提供一个简单有效的方法。     

Determination of biogenic amines in wines by HPLC with precolumn dansylation and fluorimetric detection

Thirty-two samples of commercially available Greek wines were analysed in order to determine the content of biogenic amines. The method involves pre-column dansylation of the amines and subsequent solid phase extraction (SPE) of the derivatives through C₁₈ cartridges. For the analysis, RP-HPLC (reversed phase-high performance liquid chromatography) coupled with fluorimetric detection at excitation and emission wavelengths of 320 and 523 nm, respectively was used. All amines measured had recoveries over 85%. The highest detection limit was for agmatine (0.18 mg l⁻¹). Putrescine, cadaverine, histamine and isoamylamine are the most abundant amines in the samples analysed. The relative concentrations of biogenic amines expressed in mg l⁻¹ had as follows: putrescine > histamine > isoamylamine > ethylamine > methylamine > cadaverine = tyramine = agmatine = tryptamine. Higher amounts of biogenic amines were generally detected in wines, aged for long periods in barrels or in bottles. However, young wines contained lower amounts of these compounds as they were directly bottled after winemaking and have not undergone any further maturation processing. Moreover, less acid wines gave rise to higher histamine contents.     

离子色谱-脉冲安培检测法测定原桃胶中的多糖含量

建立离子色谱-脉冲安培检测法检测桃胶中的糖类（鼠李糖、阿拉伯糖、半乳糖、葡萄糖、甘露糖）含量的方法,采用METROSEP CARB 1（150 mm×4.0 mm）分离柱,以10 mmol/L Na OH溶液在1.0 m L/min下洗脱样品溶液,总分析时间为22 min。鼠李糖、甘露糖线性范围为5.0¹00.0μg/m L,阿拉伯糖、半乳糖、葡萄糖线性范围为2.0⁶0.0μg/m L,鼠李糖、阿拉伯糖、半乳糖、葡萄糖、甘露糖检出限分别为（进样量20μL,S/N=3）0.34、0.18、0.17、0.08、0.39μg/m L,样品加标回收率为92.0%⁹8.3%,使用样品溶液连续进样6次,测得5种糖RSD为2.25%⁴.23%。此方法无需衍生、操作方便、方法快捷、检测灵敏度高,适用于桃胶类糖含量的检测。      

离子色谱法测定玄参中的单糖和低聚寡糖

建立阴离子交换色谱-脉冲安培检测法测定玄参中的葡萄糖、蔗糖、果糖和水苏糖的方法。以METROSEPCARB1(150mm×4.0mm)阴离子交换柱为分离柱,脉冲安培检测,30mmol/LNaOH溶液为淋洗液等度洗脱,超声水提法提取玄参中的糖。峰面积的相对标准偏差为0.56%～9.01%,线性范围1～50mg/L,检出限为0.0452～0.0921mg/L,相关系数R为0.9931～0.9998,加标回收率在96.52%～109.56%之间。该方法预处理简单,准确度高,适用于快速测定玄参中的单糖和低聚寡糖。     

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。     

Determination of biogenic amines in foods using ultra-performance liquid chromatography (UPLC)

A rapid ultra-performance liquid chromatographic (UPLC) method for the determination of biogenic amines (putrescine, cadaverine, spermidine, spermine, phenylethylamine, histamine, tyramine and tryptamine) in selected food samples is described. The eight biogenic amines, which are the most important to be determined in food samples, were derivatized with dansyl chloride prior to UPLC separation. The dansylated amines were separated on an Agilent Zorbax Eclipse XDB – C18 column (50 × 4.6 mm ID, 1.8 μm) using gradient elution with a binary system of acetonitrile–water, a flow rate of 1.0 ml/min and UV detection at 225 nm. The analysis is very fast, all amines are well resolved and are eluted from the column in less than 6 min. The average repeatability of the method ranged between 1.02% and 2.14%. Limits of detection (LODs) for considered amines ranged between 0.032 and 0.098 μg/l; calibration curves showed very good linearity (r = 0.9994–1.0000). The method was applied to the analysis of amines in pork, beef, chicken and fish meat, cheese and edible mushrooms.     

离子交换色谱法测定10种热带水果中的葡萄糖、蔗糖和果糖

建立了高效阴离子交换-脉冲安培检测（High performance anion exchange-pulsed amperometric detection,HPAE-PAD）测定火龙果、莲雾、牛油果等10种热带水果中葡萄糖、蔗糖和果糖的检测方法。热带水果中的多糖经超声萃取后,用METROSEP CARB 1（150 mm×4.0 mm）色谱柱进行分离,以30.0 mmol/L Na OH为流动相,等度洗脱,用安培检测器检测,18 min可完成对样品多糖的分离和定量分析。经测定,葡萄糖和蔗糖的检出限分别为0.1209μg/m L和0.2827μg/m L,线性范围为1.0⁷0.0μg/m L;果糖的检出限为0.5501μg/m L,线性范围为5.0⁸0.0μg/m L。样品溶液连续5次进样,3种糖的相对标准偏差为0.67%⁷.04%,平均回收率为78.70%¹17.75%。该方法前处理简单、选择性好、灵敏度高,可用于热带水果中可溶性糖的测定。      

Biogenic amines in meat inoculated with Lactobacillus sake starter strains and an amine-positive lactic acid bacterium

 Formation of biogenic amines in minced meat inoculated with two Lactobacillus sake starter culture strains and an amine-positive lactic acid bacterium (G-106) was studied. The effects of these starter cultures against the formation of biogenic amines were dependent on the kind of decarboxylating microorganisms present in the raw material and the effects were different for each amine. Starter strains maintained the microbiological quality of the meat kept at 20°C for 7 days, and inhibited the formation of putrescine and cadaverine. They also inhibited the formation of phenylethylamine caused by G-106 when it was present at an initial level of 10² colony-forming units (CFU)/g. However, the formation of tyramine was not affected and the formation of histamine was increased when starters were used in samples inoculated with G-106. Received: 4 December 1996 / Revised form: 27 January 1997     

Biogenic amines in meat inoculated with Lactobacillus sake starter strains and an amine-positive lactic acid bacterium

 Formation of biogenic amines in minced meat inoculated with two Lactobacillus sake starter culture strains and an amine-positive lactic acid bacterium (G-106) was studied. The effects of these starter cultures against the formation of biogenic amines were dependent on the kind of decarboxylating microorganisms present in the raw material and the effects were different for each amine. Starter strains maintained the microbiological quality of the meat kept at 20°C for 7 days, and inhibited the formation of putrescine and cadaverine. They also inhibited the formation of phenylethylamine caused by G-106 when it was present at an initial level of 10² colony-forming units (CFU)/g. However, the formation of tyramine was not affected and the formation of histamine was increased when starters were used in samples inoculated with G-106. Received: 4 December 1996 / Revised form: 27 January 1997     

发酵肉制品中生物胺及其产生菌株研究进展

生物胺是一类具有生物活性的含氨基小分子有机化合物,其广泛存在于蛋白丰富的食品中。生物体内适量生物胺具有增强代谢、促进生长等功能,但当含量达到临界值时,会产生诸多不良影响。存在大量微生物的食品中,生物胺的含量较高,同类发酵肉制品中生物胺的种类和含量也有很大不同,如发酵时间、贮藏时间和包装材料等都会对此产生影响。本文主要介绍生物胺的常见种类、生理作用,发酵肉制品中产生物胺的微生物种类、形成途径、菌株筛选方法及检测方法,旨在为今后发酵肉制品中生物胺的进一步研究提供帮助,以期为发酵肉制品的质量保障提供参考。