TLC和HPLC法相结合分析盐水鸭中的生物胺

应用薄层层析(TLC)和高效液相色谱(HPLC)两种方法分析盐水鸭中生物胺的组成及含量。薄层层析结果显示,盐水鸭样品中存在着腐胺、尸胺、亚精胺、精胺、酪胺和2-苯乙胺;利用高效液相色谱法也检测到了以上6种生物胺,且各生物胺的含量均在204μg/g以下。两种方法结果相吻合,TLC法可以作为肉制品中生物胺定性分析的一种经济便捷的方法。     

黄酒发酵液中产生物胺乳酸菌的分离鉴定与评价

从黄酒发酵液中分离得到6株乳酸菌,16S rDNA序列分析和生理生化鉴定结果表明:菌株R1、R4、R5、R8、R20为Lactobacillus rossiae,菌株R2为Lactobacillus casei。采用平板检测法对6株乳酸菌产生物胺能力进行了评价,结果显示只有乳酸菌R1具有产生物胺的能力。利用HPLC检测对平板检测结果进行了验证,结果表明乳酸菌R1能产598.61 mg/L的腐胺,其他菌株不具有产生物胺的能力。平板检测与HPLC检测结果相一致,表明平板检测可作为一种有效、操作简单、费用低的定性评价乳酸菌产生物胺能力的手段。     

一种新的柱前衍生RP-HPLC法分析酱腌菜中的生物胺

以4-fluoro-3-nitrobenzo-trifluoride(FNBT)为衍生剂,N,N-diisopropylethyl-amine(DIPEA)为催化剂,建立了一种新的柱前衍生-反高效液相色谱检测法(RP-HPLC)同时测定酱腌菜中的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)。采用硼酸盐缓冲溶液为衍生介质,在60℃下衍生反应30 min,使用C18色谱柱分离,以乙腈和超纯水为流动相梯度洗脱,流速1.00m L/min,紫外检测波长242 nm。在0.5~100 mg/L浓度范围内,8种生物胺呈现良好的线性相关(r2≥0.999),方法的检出限(LODs)为0.013~0.053 mg/L,定量限(LOQs)为0.044~0.176 mg/L,加标平均回收率为93.47%~99.87%,相对标准偏差(RSD)为0.47%~3.67%。采用该方法对26种市售酱腌菜中的生物胺进行了分析,发酵型酱腌菜中的生物胺总量范围为40.57~692.82 mg/kg,非发酵型酱腌菜中的生物胺总量范围为9.86~270.93 mg/kg。发酵型酱腌菜中8种生物胺单体的平均含量均高于非发酵型酱腌菜。该方法简单、准确和可靠,可适用于酱腌菜中生物胺的快速分析。     

工厂实测冷鲜鸡冷却贮藏过程品质的变化

探讨冷鲜鸡在冷却加工及贮藏过程中的品质变化情况。检测指标包括:p H、挥发性盐基氮(TVB-N)、菌落总数、尸胺含量、蒸煮损失率、质构指标(TPA),测试时间点为工厂环境下鸡刚宰杀、冷却排酸结束、冷藏1 d、冷藏2 d及冷藏6 d。研究表明:冷藏2 d时,鸡腿肉及鸡胸肉蒸煮损失率分别为11.86%、16.80%,为整个加工冷藏过程的最低值,且维持较好质构特性。随冷藏时间延长,鸡肉TVB-N值、菌落总数、尸胺含量均不同程度增加。冷藏6 d时,鸡腿肉TVB-N值趋近15 mg/100 g,菌落总数已略微超过一级鲜肉菌落总数值(106CFU/g),尸胺含量也达到1.820 mg/kg;而鸡胸肉TVB-N值与菌落总数均低于一级鲜肉标准值,尸胺含量为1.585 mg/kg。研究所得结论是:冷藏1 d内冷鲜鸡处于僵直期,鸡肉尚未成熟,理想食用阶段在冷藏2 d左右,且冷鲜鸡货架期不宜超过6 d。     

紫色甘薯醇提物抗疲劳机制研究

为了研究紫色甘薯醇提物的抗疲劳效应,设计小鼠运动耐力实验,检测小鼠血液生化指标和小鼠骨骼肌核蛋白PGC-1α蛋白表达的影响。结果显示,20mg/m L、10mg/m L组小鼠的运动耐力有显著性延长(P<0.01),表明紫色甘薯醇提物可以提高肌肉的耐力。生化检测结果表明,10mg/m L、20mg/m L组能降低血清尿素、丙二醛、血乳酸的含量(P<0.05),能显著增加肝糖原存储量(P<0.01),显著提高总超氧化物歧化酶的活性(P<0.01)。Western Blot检测结果显示,静止状况下,PGC-1α表达提高了3%;运动状况下,PGC-1α表达提高了6.81%。实验结果表明,紫色甘薯醇提物能提高小鼠体内抗氧化能力,增加糖原含量,降低代谢产物累积,提高PGC-1α蛋白表达来提高小鼠体内能量代谢,达到抗疲劳能力。     

黄酒酿造过程中优势细菌产生物胺的检测与评价

采用显色培养基法、脱羧酶序列特异PCR技术和HPLC分析脱羧酶液体培养基生物胺含量变化3种方法检测和评价来自黄酒酿造过程的41株细菌产生物胺的情况。研究表明,3种方法检测结果大致相同,互补使用能够准确分析细菌产生物胺的情况;41株细菌中,53.7%以上都能产酪胺,31.7%以上产腐胺,24.3%以上产组胺;14株短乳杆菌均携带酪氨酸脱羧酶基因,其中11株菌的酪胺产量超过50.00 mg/L,且多数菌能代谢产生多种生物胺,因此推断其对黄酒中生物胺含量影响最大;细菌代谢是否产生物胺及产生物胺的能力无种属特异性,但有菌株特异性。     

黄酒酿造过程中产生物胺菌株的筛选及其特性研究

从工业化黄酒酿造环境中筛选具有产生物胺能力的微生物,通过显色反应对菌株的产胺能力进行定性分析,用反相高效液相色谱(RP-HPLC)进行定量分析,获得12株产生物胺的阳性菌株。其中2株菌产生物胺含量较高,经16S r DNA序列分析,菌株AR281为短乳杆菌,菌株AR193为棒状乳杆菌。37℃时,菌株AR281总生物胺的产生量最高为154.76 mg/L;菌株AR193总生物胺的产生量最高为173.78 mg/L。本文研究了发酵温度和发酵时间对其产生物胺能力的影响,为黄酒酿造过程中生物胺含量的控制提供了理论依据。     

冰鲜鸡中主要腐败菌的分离鉴定及其产生物胺特性分析

本文用不同选择性培养基对冰鲜鸡的主要腐败菌进行分离,利用16S r RNA基因序列测定法对分离菌进行鉴定,最后利用薄层层析和HPLC法对分离菌是否代谢产生物胺进行分析。研究发现:从冰鲜鸡中共分离筛选得到11株腐败菌,分别命名为C1、C4、C7、C8、PCA1、PCA2、PCA3、PCA4、PCA5、PCA7、PCA8。PCA1与Stentrophomona(狭长平胞属)、PCA4与Kouria(考克氏菌属)、PCA3与Enterobacter asburiae(阿氏肠杆菌)、PCA7与Enterobacter cancerogenus(生癌肠杆菌)、C1与Enterobacter ludwigii(肠杆菌属)、C4与Enterobacter cloacae(阴沟肠杆菌)、C7与Enterobacter hormaechei(肠杆菌属)、C8与Enterobacter hormaechei(肠杆菌属)、PCA2与Kocuria rhizophila(克氏库克菌属)、PCA5和PCA8与Staphylococcus(葡萄球菌属)亲缘关系近。通过薄层层析初步鉴定菌是否有产生物胺能力,发现PCA2、PCA5、PCA8不产生物胺,其余菌均产生物胺。利用HPLC进一步确定产生物胺菌发酵液中生物胺组成和含量,结果 PCA7只产生腐胺,含量为844.937μg/m L;其余菌株都产含量不同的腐胺和尸胺。本文研究表明,冰鲜鸡易受肠道菌污染,且产腐胺与尸胺,为冰鲜鸡的防腐与安全食用提供一定的参考依据。      

黄酒发酵过程中乳酸菌的分离及对其产生物胺能力的评价

对黄酒酿造过程中乳酸菌进行分离,并对乳酸菌产生物胺能力进行检测。利用脱羧培养基对分离出的82株乳酸菌进行培养,结合反相高效液相色谱技术(reversed-phase high-performance liquid chromatography,RPHPLC)测定发酵液中生物胺含量,具体色谱条件如下:使用丹磺酰氯(Dansyl Chloride,Dns-CL)进行柱前衍生,以乙腈-乙腈(φ=50%)为流动相梯度洗脱,流速1.0 m L/min,254 nm紫外波检测。该方法在给定浓度范围内线性关系良好(R~20.996),平均回收率为94.46%~105.20%,相对偏差(RSD)均小于5%。检测到组胺产生菌株11株,最大生成量为18.19 mg/L,高产菌株分离自黄酒前酵第2天;酪胺产生菌株28株,最大生成量为30.38mg/L,高产菌株分离自黄酒前酵第2天;腐胺产生菌株51株,最大生成量为299.94 mg/L,高产菌株分离自浸米水。由此表明,有必要控制黄酒发酵过程中的乳酸菌,采用低产或不产生物胺的乳酸菌作为强化菌株,可降低黄酒中的生物胺含量。     

二十二碳六烯酸海藻糖酯的分离纯化及鉴定

在叔丁醇中以二十二碳六烯酸(DHA)乙酯和海藻糖为原料,通过固定化脂肪酶催化合成DHA海藻糖酯,并对其分离纯化方法进行研究。确定DHA海藻糖酯的薄层层析(TLC)条件:展开剂为乙酸乙酯/甲醇/水(8.5/1/0.5,v/v/v),碘蒸气显色10min;硅胶柱层析条件:正己烷/异丙醇/甲醇(5/4/1和4/4/2,v/v/v)梯度洗脱,流速1.3mL/min,1管/7min收集洗出液;用半制备高效液相色谱(HPLC)进一步纯化单酯收集液,经分析型HPLC检测纯度后用核磁共振(NMR)方法进行结构鉴定,确定为DHA藻糖单酯。     

冷藏兔肉中嗜冷菌的分离鉴定及其产生物胺的特性分析

从冷藏兔肉中分离出10 株嗜冷菌,用16S rDNA对菌株进行鉴定。B25为沙雷氏菌（Serratia）,其他9 株为假单胞菌（Pseudomonas）。采用薄层层析法检测10 株菌的产生生物胺能力,检测到B25可产生多种生物胺,其他9 株没产生生物胺。将B25菌接到加氨基酸前体的假单胞菌肉汤培养基中培养24 h,经高效液相色谱法分析该菌株产生生物胺的种类和含量。最终测得该菌株可产生腐胺、苯乙胺和尸胺,产生这3 种生物胺的能力均比较强,测得腐胺的含量为0.41 mg/mL,尸胺的含量为0.23 mg/mL,苯乙胺的含量为0.05 mg/mL。因此,该菌对冷藏兔肉产品的质量安全威胁较大。     

江西产腐乳生物胺水平调查及产胺菌的分离与鉴定

采用高效液相色谱法对江西产腐乳生物胺含量水平进行调查分析,通过微生物学分离结合高效液相色谱确认,筛选出生物胺产生菌,并对其进行鉴定。结果表明,12种腐乳样品中总生物胺含量范围为68.5～1084.0 mg/kg,色胺、腐胺、组胺和酪胺是主要生物胺,其中腐胺和酪胺的含量较高。筛选出的9株产胺菌产生物胺的含量范围为93.8～369.2 mg/L。对产胺量较高的6株菌进行表型鉴定和基因型16S rDNA序列测定,得到耐热芽孢杆菌（Bacillus sporothermodurans）1株,鲁梅利杆菌（Rummeliibacillus stabekisii）1株,耐硼赖氨酸芽孢杆菌（Lysinacillus boronitolerans）3株和皮脂葡萄球菌（Staphylococcus piscifermentans）1株。研究结果对腐乳食用安全性评价提供了理论依据,为进一步探清如何抑制腐乳中产胺菌的活性提供了理论研究基础。     

不同培养条件及群体感应信号分子对蜂房哈夫尼亚菌生物被膜的影响

为研究蜂房哈夫尼亚菌(Hafnia alvei)Ha-01生物被膜形成的过程及不同培养条件(碳源、p H值、Na Cl质量分数、黏附材料)对其生物被膜形成的影响,采用超声波平板法及扫描电镜法研究不同培养条件下菌株Ha-01生物被膜活菌数及生长情况,并通过添加外源标准群体感应信号分子(N-酰基高丝氨酸内酯(N-acyl-homoserine lactones,AHLs))中的C6-HSL研究了AHLs与其生物被膜形成的关系。结果显示,菌株Ha-01生物被膜的形成与培养时间密切相关;在不同碳源的培养条件下形成生物被膜能力不同,其中在以木糖为培养基时形成量最大,达到7.51(lg(CFU/cm~2)),与在LB培养基中相比增加10.28%;在中性培养条件下更利于其生物被膜的形成,活菌数为7.77(lg(CFU/cm~2));在Na Cl质量分数为2%时,其生物被膜产生量最大,活菌数为7.18(lg(CFU/cm~2));在不同黏附材料上生物被膜形成能力从大到小依次为铝片、锌片、玻璃片,活菌数分别为7.22、6.48、6.11(lg(CFU/cm~2));且添加C6-HSL量越多,其生物被膜产生能力越强。研究表明,培养条件能够影响菌株Ha-01生物被膜形成,且AHLs可以调控其生物被膜形成。     

Gas formation in ground beef chubs due to Hafnia alvei is reduced by multiple applications of antimicrobial interventions to artificially inoculated beef trim stock

Gas-forming microorganisms were isolated from gas-swollen ground beef chubs obtained from a commercial source and were phenotypically identified as Hafnia alvei. In in situ experiments, the isolated H. alvei strains produced gas in inoculated irradiation-sterilized ground beef chubs. A five-strain cocktail of H. alvei isolates was inoculated on beef trim. The inoculated beef trim samples were treated with either a water wash (W) at 65 psi for five passes (a pass refers to the application of successive multiple antimicrobial treatments to inoculated beef trim on a moving processing conveyor belt at a speed of 1 cm/s under heat ducts or oscillating spray nozzles), W plus a 2% (vol/vol) lactic acid wash (L) at room temperature at 30 psi for three passes (W/L), or a combination treatment (COMB) consisting of W plus 82 degrees C water for three passes plus 510 degrees C hot air for six passes plus L, or were not treated (control). After treatment, the beef trim was ground and vacuum packaged. The numbers of H. alvei were reduced with water alone and with the aforementioned antimicrobial intervention treatments. For the untreated and inoculated control samples, the numbers of H. alvei increased from 7.03 to 8.40 log CFU/g after 7 days of incubation at 4 degrees C. However, the numbers of H. alvei treated by successive antimicrobial interventions (COMB) were initially reduced to 5.25 log CFU/g and increased to just 6.9 log CFU/g after 7 days of incubation at 4 degrees C. Gas was produced in untreated control samples after 3 days at 15 degrees C (15 of 15 inoculated chubs). However, in meat treated with W, W/L, and COMB, gas was produced after 4 to 5, 7 to 8, and 9 to 10 days of storage at 15 degrees C, respectively. These results demonstrate the effectiveness of multiple antimicrobial interventions in reducing H. alvei numbers on beef trim and subsequently delaying gas formation in the resulting ground beef chubs.     

鱼露中组胺降解菌的分离筛选及耐受性

生物胺(biogenic amines)在某些食品尤其是发酵食品中广泛存在,具有一定的食用安全隐患.为获得用于鱼露等发酵食品的生物胺降解菌,从天然发酵鱼露中采用双层显色培养基法初步筛选出不产生物胺的菌株,再用高效液相色谱(HPLC)法进行复筛,得到一株具有高效组胺降解能力的菌株MZ5.经鉴定该菌株为库德毕赤酵母(Pic...     

Photobacterium phosphoreum caused a histamine fish poisoning incident

An incident of histamine fish poisoning (HFP) occurred due to the consumption of iwashi maruboshi (dried sardine) in Osaka, Japan in March 2002. A histamine-producing bacterial strain, YS4-7, was isolated from iwashi maruboshi that contained 1700 mg of histamine per kilogram. This strain was identified as Photobacterium phosphoreum by biochemical examinations and partial sequencing of 16S rDNA. P. phosphoreum YS4-7 showed greater capability as a histamine producer at 4 and 12 degrees C than Morganella morganii JCM 1672. Strain YS4-7 produced 546 mg of histamine per kilogram in a sardine homogenate stored for 12 h at 20 degrees C. M. morganii, Raoultella planticola and Hafnia alvei have been isolated from fish implicated in HFP incidents, whereas this is the first report of P. phosphoreum being the causative bacterium in a sporadic case of histamine food poisoning.     

产赤藓糖醇菌株的筛选与鉴定

利用含有300g/L 葡萄糖的高渗培养基从蜂蜜、花粉、土壤等样品中筛选耐高渗酵母菌,经薄层层析和高效液相色谱分析得到两株产赤藓糖醇且不产甘油的酵母菌,通过高碘酸氧化法筛选出其中赤藓糖醇产量较高的一株菌株E54。菌株E54 在含葡萄糖200g/L、酵母膏5g/L 的发酵培养基中发酵90h,赤藓糖醇产量为41.1g/L,转化率为22.8%。通过形态观察、生理生化实验、5.8S rDNA 序列分析并构建系统进化树,初步鉴定E54 为Moniliellaacetoabutans(丛梗孢酵母)。     

Formation of biogenic amines by Gram-negative rods isolated from fresh,spoiled, VP-packed and MAP-packed herring (<Emphasis Type="Italic">Clupea harengus</Emphasis>)

Bacterial strains (120) were isolated from fresh, spoiled, VP-packed and MAP-packed herring. Identified bacterial strains were investigated for their abilities to produce biogenic amines in histidine, lysine and ornithine decarboxylase broth by a rapid high-performance liquid chromatography (HPLC) method. The microflora of fresh herring was dominantly Pseudomonas (30%), Enterobacteriaceae (23.2%), Vibrio (13.3%) and Moraxella (13.3%) but, the microflora of herring stored in VP and MAP was dominated by species belonging to Vibrio (23.3%) and Moraxella (20%), which indicates that these packaging systems prevented the growth of Pseudomonas and Enterobacteriaceae. In a laboratory medium containing amino acid (histidine, ornithine and lysine), most of bacterial strains produced histamine, putrescine and cadaverine. The highest histidine decarboxylase activities were observed in Klebsiella oxytoca, Hafnia alvei and Proteus vulgaris which produced 396, 232 and 54 mg histamine/L, respectively in histidine-enriched broth. The accumulation of cadaverine by Klebsiella oxytoca and Hafnia alvei was 325 and 208 mg/l, respectively. All strains isolated produced putrescine in an ornithine-enriched broth, ranging from 3 to 249 mg/l. The production of putrescine by Klebsiella oxytoca and Hafnia alvei was 249 and 195 mg/l, respectively. Moraxella spp and Acinetobacter spp did not produce histamine which indicates they did not have histidine decarboxylase activity.