利用CO2超临界二次萃取方法提取玫瑰精油

玫瑰采摘后用乙醇浸提、浓缩,利用CO2超临界二次萃取方法萃取高质量的玫瑰精油,同时回收乙醇。结果表明：玫瑰粗油得率约为0．35％,精油的得率为0．10％-0．12％,同时乙醇的回收率约为23％～25％;获得的玫瑰精油质量符合FCC第V版要求。使用色谱／质谱联用仪分别对CO2超临界萃取玫瑰精油的成分进行了初步分析,结果分别测出香茅醇等成分42种。     

荞麦籽粒乙醇提取物抗氧化成分的高效液相色谱电喷雾质谱研究

 【目的】研究甜荞麦和苦荞麦籽粒乙醇提取物中的主要抗氧化成分。【方法】将荞麦籽粒乙醇提取物与二苯基苦基苯肼（DPPH）自由基反应,然后用高效液相色谱仪识别乙醇提取物中的抗氧化成分,并用电喷雾质谱检测器对抗氧化成分的结构信息进行表征,以确定荞麦籽粒乙醇提取物中的抗氧成分。【结果】苦荞麦和甜荞麦籽粒乙醇提取物对DPPH自由基均有清除作用;高效液相色谱分析表明,甜荞麦和苦荞麦乙醇提取物中均含有两种主要抗氧化峰,且对应的色谱保留时间和光谱信息相同;电喷雾质谱分析表明,荞麦籽粒乙醇提取物中抗氧化成分的分子量和质谱碎裂行为分别与对照品芦丁和槲皮素的相同。【结论】采用HPLC-MS/MS法可以快速筛选荞麦籽粒提取物中的抗氧化成分,荞麦籽粒提取物中的抗氧化成分主要为芦丁和槲皮素,其中槲皮素的抗氧化活性高于芦丁。     

利用CRISPR/Cas9和λ-Red级联技术构建产肠毒素大肠杆菌LT敲除菌株

本研究旨在利用CRISPR/Cas9和λ-Red级联的技术对产肠毒素大肠杆菌（enterotoxigenic Escherichia coli,ETEC）K88的热不稳定性肠毒素（heat-labile toxin,LT）基因进行无痕敲除并获得K88 LT^-缺陷菌株。通过序列比对获取LT两端同源序列,并构建包含LT边界、氯霉素筛选标记、sgRNA和LT同源臂的供体片段;将供体片段转化至ETEC K88,同时分别利用λ-Red同源重组系统和CRISPR/Cas9基因编辑系统,对LT基因进行敲除;通过PCR验证获得了K88 LT^-缺陷菌株,并通过试验测定了敲除菌株的溶血能力和生长曲线。结果显示,λ-Red同源重组系统可成功地将LT基因替换为相应的供体片段,CRISPR/Cas9基因编辑系统可高效地对筛选标记进行删除,最终通过λ-Red和CRISPR/Cas9结合的基因编辑系统可成功对ETEC K88的LT基因进行无痕敲除。体外试验结果表明,K88 LT^-缺陷菌株的溶血能力丧失,并且生长速度比野生型菌株减缓,LT可能和ETEC K88的致病能力和生长性能有关。表明λ-Red和CRISPR/Cas9级联的基因敲除方法可用于LT毒素基因及其他一些大肠杆菌基因的敲除。K88 LT^-缺陷菌株的构建为下一步研究LT毒素的致病机制奠定基础。     

Microbial properties and litter and soil nutrients after two prescribed fires in developing savannas in an upland Missouri Ozark Forest

On some landscapes periodic fire may be necessary to develop and maintain oak-dominated savannas. We studied the effects of two annual prescribed burns to determine their effect on microbial activity and soil and litter nutrients 1 year after the last burn. Surface litter and soil from the upper 0–5 cm soil layer in three developing savannas (oak-hickory, Quercus-Carya), oak-hickory-pine (Quercus-Carya-Pinus), and pine (Pinus) were collected one year after the second of two annual prescribed burns. Surface litter was analyzed for nutrients and soil was analyzed for phospholipid fatty acids (PLFAs) and nutrients. Surface litter chemistry differed across the three savannas for potassium (K) and boron (B), being significantly (P < 0.05) higher for unburned forest than for burned forest. Among savannas, only sulfur (S) was higher for the pine savanna and B for the oak-hickory savanna, both were higher for unburned forest than for burned forest. For soil, calcium (Ca) and B differed across savannas, being higher for burned forest than for unburned forest. Among savannas, soil pH, Ca, and B concentrations were higher in soil from burned forest than from unburned forest. Total PLFA differed among savannas, but was not affected by burning treatments. However, the amounts of biomarkers for Gram-positive and Gram-negative bacteria were higher while the amount of biomarker for fungal PLFA was lower for burned forest than for unburned forest. Our results indicate that the two annual prescribed burns moderately affected PLFA microbial community structure and litter and soil nutrient concentrations. However, the long-term effects of fire on these study sites are not known and merit further study.     

超高效液相色谱串联质谱法同时检测复杂基质中四环素类抗生素及其代谢产物

为同时检测复杂基质中（由猪粪和蘑菇渣混合而成的堆肥原料）3种四环素类抗生素（四环素TC、土霉素OTC和金霉素CTC）及其代谢产物,建立了超高效液相色谱串联质谱检测方法（UPLC—MS／MS）。该方法同时采用pH值=4的Na2EDTA-Mellvaine缓冲溶液和乙腈为提取溶液,经过固相萃取净化后,以乙腈和0．1％的甲酸水溶液为流动相,采用超高效液相色谱柱进行分离,在电喷雾正离子模式下,用四极杆串联质谱仪进行定性和定量分析。3种四环素类抗生素及其代谢产物均在7min内完成分离,总共分析时间为12min。在0-6mg·kg^-1DW（Dryweight）浓度范围内,3种四环素类抗生素及其代谢产物的标准曲线线性良好,线性相关系数R2均大于0．9960,重现l生也较好（n=11,相对标准偏差均小于15％）。在3个加标水平0．2mg·kg^-1DW、1mg·kg^-1DW和4mg·kg^-1DW下,TC、OTC、CTC的回收率分别为71％-89％、66％～94％和66％～84％;去甲基金霉素（DMCTC）的回收率为52％-64％;差向异构产物的回收率在32％～51％之间;脱水产物以及差向脱水产物的回收率均低于30％。3种四环素类抗生素及其代谢产物的检出限和定量限分别在1．668～17．270μg·kg^-1和5．561—45．918μg·kg^-1范围内,表明该方法具有较高的灵敏度。对北京市某露天堆肥场中的样品进行测定发现,TC、OTC、CTC的浓度分别为0．4、1．6、2．9mg·kg^-1,检测到的代谢产物主要为相应的差向异构体,其中差向金霉素（ECTC）的浓度最高,达到2．7mg·kg^-1,和母体的含量水平比较接近,其他代谢产物也有不同程度的检出。     

纳米ZnO对浮萍（Lemna minor L.）的生态毒性及其聚集性和溶解性分析

纳米颗粒的大量生产和应用增加了其环境释放风险,为此以不同浓度的纳米ZnO（0、1、10mg·L-1和50mg·L-1）处理浮萍（LemnaminorL.）7d,分析了纳米颗粒对植物D665/D665a值（叶绿素与脱镁叶绿素的比率）以及超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、过氧化物酶（POD）和Na＋K＋-ATP酶（Na＋K＋-ATPase）活性的影响,并对纳米ZnO的聚集性与溶解性进行了测试。研究结果显示,浓度为50mg·L-1的纳米颗粒显著抑制D665/D665a值和Na＋K＋-ATPase活性,而抗氧化酶活性则显著升高;纳米颗粒在培养液中易发生聚集作用而沉积,12h后基本上完全沉积到底部。结果说明,50mg·L-1的纳米ZnO对浮萍产生了显著的胁迫作用,其对浮萍的毒性作用主要来源于其溶出的Zn2＋。     

缓控释氮素肥料的研究现状与展望

在前人有关缓控释肥料文献的基础上,介绍了缓控释氮肥的缓效途径和研究发展情况。重点阐述了缓控释氮肥的分类和缓控释机理。此外,对今后缓控释肥料研究发展提出了一些见解。     

超高效液相色谱—串联质谱法测定牛奶中6种玉米赤霉烯酮类霉菌毒素的残留

试验旨在建立牛奶中6种玉米赤霉烯酮类霉菌毒素残留的超高效液相―串联质谱检测方法。牛奶样品经乙腈提取,免疫亲和柱净化,C₁₈色谱柱分离,以0.02%（V/V）乙酸―水/甲醇为流动相,采用梯度洗脱方式进行液相色谱分离,多反应监测模式,外标法进行定量。牛奶中6种玉米赤霉烯酮类霉菌毒素的检测限可达到1.0 μg/L,且在2.5～100.0 μg/L的浓度范围内,线性关系良好。该方法专属性强、灵敏度高、分离度好,适用于牛奶中的6种玉米赤霉烯酮类霉菌毒素残留的测定。     

Effects of Momordica grosvenori Saponins on Type 2 Diabetes Rats Induced by High-sucrose/High-fat Diet and Streptozotocin

The purpose of the experiment was to study the effect of Momordica grosvenori saponins on improving the effect of high-sucrose/high-fat diet combined with streptozotocin induced type 2 diabetes in rats.The type 2 diabetes rat model was established with a high-sucrose and high-fat diet combined with streptozotocin.Successfully modeled rats were given the total saponins of Momordica grosvenori (100,200,400 mg/kg) or metformin (268 mg/kg) by gavage,once a day for 4 weeks.Recording the "three more and one less"index (24 h internal urine output,water intake and feed intake,body weight);Detecting of biochemical indicators,analysis of fasting blood glucose (FBG),serum total cholesterol (TC),triglycerides (TG),low-density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol (HDL-C),insulin (INS),catalase (CAT),superoxide dismutase (SOD),malondialdehyde (MDA),alanine aminotransferase (AST),aspartate aminotransferase (ALT),blood urea nitrogen (BUN) and creatinine (Cr) levels.Hematoxylin-eosin staining (HE) was used to observe the pathological changes of the pancreas.Compared with the model group,the feed intake,water intake,urine output,blood indicators FBG,INS,MDA,TC,TG,BUN and Cr levels,AST and ALT activities were significantly or extremely significantly decreased in the total saponins of Momordica grosvenori each dose group and metformin group (P<0.05;P<0.01),while CAT,SOD activities and HDL-C level were significantly or extremely significantly increased (P<0.05;P<0.01).In pancreatic tissue,the number of pancreatic cells was increased significantly,and the pathological damage of the pancreas was significantly reduced. Momordica grosvenori saponins and metformin had obvious effects on improving glucose metabolism disorders,and at the same time could significantly reduce insulin resistance,enhance the body’s ability to resist oxidative stress and improve liver and kidney functions.