不同茶叶中游离氨基酸的对映异构体

【目的】游离氨基酸是茶叶的主要化学成分之一,与茶叶的滋味品质密切相关。大多数氨基酸具有手性中心,因此存在滋味特性及生理活性均可能截然不同的L型和D型两种对映异构体。由于检测技术的限制,过去的研究多集中在L型氨基酸上,而茶叶中D型氨基酸罕有系统的研究报道。因此,研究茶叶中游离氨基酸对映异构体,对茶叶化学理论体系的深化及滋味品质的提升与调控具有重要的推进作用。【方法】采用手性HPLC法、MTBSTFA硅烷衍生化法和酯化-PFP酰化法3种不同的分析方法对比茶叶中游离氨基酸对映异构体的分离性能,结果表明采用酯化-PFP酰化法结合手性气相色谱-质谱联用技术（gas chromatography-mass spectrometry,GC-MS）,可实现茶叶中丙氨酸、缬氨酸、苏氨酸、脯氨酸、亮氨酸、异亮氨酸、丝氨酸、半胱氨酸、天冬氨酸、蛋氨酸、苯丙氨酸、谷氨酸、酪氨酸、赖氨酸及色氨酸等15种重要游离氨基酸对映异构体的有效分离。继而建立茶叶中游离氨基酸对映异构体的高效衍生化及定性定量分析方法,并查明市售11种代表性白茶、乌龙茶及普洱茶中游离氨基酸对映异构体的分布规律。【结果】衍生化条件为：将氨基酸反应液在100℃下酯化105 min,继而以四氢呋喃为反应溶剂,在100℃下五氟丙酰化10 min。游离氨基酸对映异构体的回收率介于75.26%—123.6%（低浓度）、81.23%—121.8%（高浓度）,RSD值介于2.09%—13.12%（低浓度）、1.48%—10.59%（高浓度）。茶样分析结果表明,大部分氨基酸的1—2个对映异构体可以在茶样中检测出,且D-氨基酸普遍存在于茶样中,尤其是D-苏氨酸、D-天冬氨酸、D-半胱氨酸、D-苯丙氨酸、D-茶氨酸、D-谷氨酰胺及D-谷氨酸混合物在大部分茶样中均有分布,而D-蛋氨酸、D-赖氨酸及D-色氨酸在所有茶样中均未被检测出。在具体含量分布上,除了L-茶氨酸、L-谷氨酸及L-谷氨酰胺混合物之外,L-半胱氨酸（1.48—2.08 mg?g^-1）、D-半胱氨酸（1.46—1.49 mg?g^-1）、L-丝氨酸（0.15—1.80 mg?g^-1）及D-天冬氨酸（1.02—1.14 mg?g^-1）在白茶中的含量普遍较高;L-半胱氨酸（1.52—1.70 mg?g^-1）、D-半胱氨酸（1.45—1.49 mg?g^-1）、L-丝氨酸（1.03—1.50 mg?g^-1）、L-蛋氨酸（1.03—1.52 mg?g^-1）、L-酪氨酸（1.32—1.35 mg?g^-1）及D-天冬氨酸（1.01—1.15 mg?g^-1）在乌龙茶中具有较高含量。在普洱茶中,L-茶氨酸、L-谷氨酸及L-谷氨酰胺混合物的含量（1.04 mg?g^-1）与其他氨基酸相比并无显著性差异,而L-苏氨酸、L-色氨酸及L-丝氨酸含量较高,介于0.61—0.84 mg?g^-1。在氨基酸总量上,各茶类呈现出白茶（最高为40.61 mg?g^-1）>乌龙茶（最高为25.43 mg?g^-1）>普洱茶（8.01 mg?g^-1）的分布规律。多元统计分析结果表明,L-茶氨酸、L-谷氨酸及L-谷氨酰胺混合物、L-色氨酸及L-天冬氨酸在部分白茶中的含量显著高于乌龙茶,而L-酪氨酸与L-蛋氨酸则在乌龙茶中含量较高。【结论】D-氨基酸存在于大部分茶叶中,对茶汤滋味品质存在潜在影响,但在本研究样例中,D-氨基酸的含量分布在不同茶类间未表现出显著性差异。     

液液萃取-QuEChERS净化-高效液相色谱-串联质谱法测定饲料添加剂可溶性粉中五氯酚酸钠含量

建立液液萃取-QuEChERS净化-高效液相色谱-串联质谱法测定可溶性粉中五氯酚酸钠含量的新方法。 可溶性粉先用水溶解后加入1moL/L 盐酸1mL调pH至4左右,加入乙醚液液萃取到乙醚层,蒸干乙醚,乙腈复溶,QuEChERS净化管净化后高效液相色谱-串联质谱测定,标准曲线外标法定量。 五氯酚酸钠在0.50~100μg/L质量范围内线性关系良好,相关系数为0.9994,检出限0.5μg/kg,定量限1.0μg/kg,可溶性粉剂中五氯酚酸钠添加量为1.0、5.0、10.0μg/kg时,五氯酚酸钠的平均回收率为80.2%~94.0%,相对标准偏差小于7%。 该方法快速、高效且价格低廉,适用于饲料添加剂可溶性粉中五氯酚酸钠的快速筛查分析测定。     

养猪废水恶臭挥发性物质释放特征及其组分源解析

为探究猪场曝气池和出水口废水水样的恶臭挥发性物质（Malodorous Volatile Compounds,MVCs）组成和释放特征,本文采用质子转移反应质谱（PTR-MS）检测了养猪废水在实验室模拟存储过程中顶空MVCs组成,检出了含苯芳香烃、胺类、硫化物、酮类、酚类、羧酸和吲哚7类共22种MVCs。两种废水中MVCs浓度较高是氨、硫化氢、丙酮、二甲胺、甲基乙基酮、甲硫醇和丙酸等。通过臭味活度值（OAV_i）分析了两种废水的臭气组分相对贡献率,曝气池废水释放的MVCs臭味贡献度较高的是甲硫醇（28.77%）、3-甲基吲哚（19.05%）、对甲苯酚（18.45%）、硫化氢（15.05%）和二甲胺（7.15%）;而出水口液样释放的MVCs臭味贡献度较高的是对甲苯酚（47.17%）、甲硫醇（38.26%）和3-甲基吲哚（14.57%）。胺类和羧酸类物质在整个储存过程中的浓度变化较小,硫化物在所有挥发性有机物中所占的比例较低,在存放过程中浓度变化差异较小。曝气池废水中释放速率最高的MVCs是氨、丙酮和二甲胺,分别为47 928、27 μg·h^-1·m^-2和15 μg·h^-1·m^-2。主成分分析结果显示MVCs组分来源为4类,分别为含碳有机物的厌氧消化、蛋白质分解、粪浆及冲刷水中的颗粒物和动物排泄。     

三种消解方法测定天然橡胶镁含量的研究

为研究更适合于天然橡胶中镁含量检测的方法,采用干灰化法、微波消解法和石墨消解法三种消解方法研究不同消解条件对天然橡胶中镁含量检测的影响。结果表明,三种消解方法对天然橡胶镁含量的测定均可以满足要求。干灰化法对实验结果影响较小,对低镁含量的样品应适当增加样品量;微波消解法操作简便且检测耗时较短;石墨消解法操作简单,工作效率较高,适用于批量样品分析,设备以及耗材成本与前两种方法相比具有较大优势,为未来我国制定天然橡胶中镁含量的检测标准提供了理论依据。     

海产品中溴酚类海洋风味素的气相色谱—质谱联用分析

建立了海产品中的溴酚类海洋风味素的毛细管气相色谱-质谱联用检测法。样品中溴酚类化合物在挥发油测定器中用乙酸乙酯通过循环蒸馏萃取,浓缩后用电子轰击源在特征离子碎片扫描检测模式下进行检测。结果表明,5种溴酚类化合物在SBP-50柱上得到良好的分离,色谱出峰尖锐,没有杂峰干扰。本方法在2.5～40 mg·L~(-1)浓度范围内线性关系良好,最低检测限低于1ng·mL~(-1)。当向25.00g样品中添加溴苯酚各2.0μg时,平均回收率分别达到90.45％(2-BP)、60.49％(4-BP)、85.79％(2,4-DBP)、84.98％(2,6-DBP)、90.80％(2,4,6-TBP),标准相对偏差为1.37％～4.16％。对市场上销售的几种海洋经济动物测定,溴酚类风味素含量在5.40～31.38 ng·g~(-1)范围内,而所测得的淡水青虾中溴酚类风味素含量只有0.18 ng·g~(-1)。     

Hepatic emphysema associated with ultrasound-guided liver biopsy in a dog

An eleven-year-old Chinese Crested Powder Puff dog presented with polydipsia/polyuria, inappetence, diarrhea and vomiting underwent an ultrasound-guided percutaneous liver biopsy. Two days post-biopsy the clinical condition of the dog acutely deteriorated with fever, dyspnea, ataxia and subcutaneous emphysema. Radiographs and ultrasound showed focal severe hepatic emphysema in the region of the previous liver biopsy. Post-mortem examination revealed chronic hepatitis with dissecting fibrosis, acute hepatitis with hemorrhage and in the hindlimb musculature extensive hemorrhage and necrosis. Pure cultures of the gas producing bacteria Clostridium perfringens were isolated in samples from the hind limb musculature. We propose that the hepatic emphysema in the region of the biopsy site was a result of a clostridial infection.     

Investigation of the Role of Androstenedione-19-oic Acid in the Presence of 19-Norandrostenedione in Intact Male Horse Plasma Using Liquid Chromatography–Tandem Mass Spectrometry

A liquid chromatography–tandem mass spectrometry method was developed to confirm the presence of androstenedione-19-oic acid in intact male equine plasma and to show the source of 19-norandrostenedione in equine plasma. Androstenedione-19-oic acid was recovered from acidified plasma by liquid–liquid extraction using methyl tert-butyl ether and separated on an Ace 5 C₈ column. A triple quadrupole mass spectrometer was used to detect the analytes in negative electrospray ionization mode. Limits of detection, quantification, and confirmation of the method were 0.1, 0.5, and 1.0 ng/mL, respectively. The linear dynamic range of quantification was 0.5–50 ng/mL. The presence of androstenedione-19-oic acid was confirmed in all plasma samples obtained from intact male horses but not those from gelded and female horses; the average concentration was 3.1 ± 1.6 ng/mL, suggesting androstenedione-19-oic acid is an endogenous compound only in intact male horse plasma samples. The conversion of androstenedione-19-oic acid to 19-norandrostenedione in equine plasma was demonstrated by spiking androstenedione-19-oic acid into blank plasma and monitoring the generation of 19-norandrostenedione and its increase in concentration during storage. Results indicated that androstenedione-19-oic acid was readily converted into 19-norandrostenedione; the higher the storage temperature, the faster the conversion. The conversion was not affected by the types of plasma samples collected from gelded and female horses or by anticoagulants used in blood collection to harvest plasma. Compared with other matrices such as water, methanol, and phosphate-buffered saline, the conversion of androstenedione-19-oic to 19-norandrostenedione in equine plasma was faster, suggesting that there is an unknown factor(s) in equine plasma that enhances the conversion.     

氢化物发生－原子荧光光谱法测定止痢散中三氧化二砷的含量

为完善止痢散质量标准,采用氢化物发生-原子荧光光谱法对止痢散中三氧化二砷的含量测定进行了研究。载流为3．0％盐酸溶液,还原剂为含0．5％氢氧化钾的1％硼氢化钾溶液,氩气载气流量为600mL／mL,砷空心阴极灯电流为60mA。砷元素进样浓度在0～100ng／mL范围内与荧光强度值呈良好的线性关系（r=0．9999）;样品平均回收率为95．3％（n=6）,RSD为1．9％。本方法简便、准确度高、重复性好,为进一步控制止痢散的质量提供了依据。     

Characterization of hepatic and extrahepatic glutathione S-transferases in rainbow trout (Oncorhynchus mykiss) and their induction by 3,3′,4,4′-tetrachlorobiphenyl

Liver, kidney, gill and olfactory epithelium cytosolic fractions of rainbow trout (Oncorhynchus mykiss) were examined for glutathione S-transferase (GST) contents. Proteins retained on a glutathione (GSH)-affinity matrix were separated as monomers by reversed-phase HPLC and characterized by immunoblotting, mass spectrometry and partial amino acid sequence. For each organ concerned, a specific pattern of these proteins was determined and appeared similar for liver and kidney on one hand, and for gill and olfactory epithelium on the other hand. It was confirmed that the prominent hepatic GST is a class enzyme, also constitutively expressed as a major isoform in the four organs studied. Moreover, a class variant and two new class GST subunits were characterized in minor fractions. An unknown protein, which was found major in gills and olfactory epithelium, exhibited some characteristics of class GSTs. Occurrence of possible GSH-adduct formation observed on two distinct monomers in specific experimental conditions is discussed. These results and methods were used to investigate the effect of 3,3,4,4-tetrachlorobiphenyl (TCB), a polychlorinated biphenyl (PCB), on GST expression in trout liver. From HPLC-profiling, significant co-induction of the major class and the two minor class GST subunits was observed in trout after waterborne exposure to TCB which was followed by a slight increase in 1-chloro-2,4-dinitrobenzene (CDNB) activity. The present work allows qualitative evaluation of the specific detoxification potential of rainbow trout. The use of HPLC-profiling of GSTs as a possible tool for the biomonitoring of polluted aquatic environment is suggested.     

液相色谱-串联质谱法测定水产品中孔雀石绿的残留量

建立了液相色谱-串联质谱测定水产品中孔雀石绿及其代谢物残留的检测方法。样品经乙酸铵盐溶液和乙腈提取,经二氯甲烷萃取,PRS阳离子交换柱净化,然后选用电喷雾离子源,在正离子、多反应监测方式模式下进行定性,以内标法进行定量。方法的检出限为0.10μg·kg-1,工作曲线的线性范围为0.10~4.00μg·kg-1。在添加浓度为0.10~4.00μg·kg-1的范围内,孔雀石绿和隐色孔雀石绿的平均回收率均分别为83.4%和95.0%,相对标准偏差分别为6.23%和6.74%。