1、专业可行性报告作者简介: 刘美娜(1982) ,女,汉族,山东烟台人,烟台南山学院纺织服装学院,教师。罗布麻纤维有效成分抑菌机理初探刘美娜(烟台南山学院,山东烟台,265713)摘要:采用烧瓶振荡法测得罗布麻纤维、提纯后的罗布麻纤维的对大肠杆菌抑菌率分别为 57%、27%,对金黄色葡萄球菌的抑菌率分别为 34%、21.7%。经过提纯后罗布麻纤维的抑菌率明显下降,这说明罗布麻纤维中含有抑菌成分,但通过质谱分析,罗布麻纤维中并不含黄酮类化合物关键词:罗布麻纤维; 黄酮类化合物; 质谱分析; 抑菌中图分类号:F810.42 文献标识码:AThe study on antibacterial mech
2、anism of the active substances extracting from the Apocynum Venetum fiber (AV fiber)Liu Meina (Yantai Nanshan Unversity, Yantai, Shandong, 265713)Abstract: In this paper, flask-shaking method is used to experiment on the antibacterial ratios of AV fibers and extracted AV fibers, which separately are
3、 57%, 27% to colibacillus, 34%, 21.7% to staphylococcus. The MS analysis results indicate the disappearance of flavone.Key words: AV fibers; flavone; MS analysis; antibacterial目前,临床和试验结果都表明罗布麻纤维具有抑菌性,但目前其抑菌机理尚不清楚。罗布麻是一种中草药,其叶、茎、根、花中均含有黄酮类、强心甙、甾醇、鞣质、酚类、萜类等中药成分 1,2,这些物质对细菌有一定的抑杀作用 3。罗布麻纤维由罗布麻麻皮脱胶制得,所以
4、现在普遍认为它的抑菌性主要是由于其含有的黄酮类化合物所致。本文通过烧瓶振荡法 4,5来研究罗布麻纤维中有效成分的抑菌性能,并通过质谱分析法来确定罗布麻纤维中含有的黄酮类化合物。专业好文档1 实验1.1 实验材料与菌种 棉纤维和罗布麻纤维;菌种包括大肠杆菌和金黄色葡萄糖球菌。棉纤维煮炼(NaOH 1.5%,3h)水洗酸洗水洗中和水洗漂白(H 2O2 3 g/L4 g/L)热水洗水洗烘干检验罗布麻纤维:煮炼(NaOH 2%,2h)水洗酸洗水洗中和热水洗水洗烘干提取罗布麻纤维:煮炼(NaOH 2%,2h)甲醇回流提取(水浴加热 60,4h)乙醇回流提取(水浴加热 70,2h)80%乙醇回流提取(水浴
5、加热80,2h)热水洗(6080)水洗烘干罗布麻麻皮/罗布麻纤维(各 8 克)甲醇回流提取(水浴加热 60,6h,2h 更换一次甲醇)浓缩(10ml)1.2 试验方法与仪器抗菌实验按照新的纺织行业标准FZ/T73023-2006抗菌针织品,采用烧瓶振荡法。使用的仪器为GSP-9270MBE隔水式恒温培养箱,上海博讯实业有限公司;SHZ-82恒温振荡器,广州国华电器有限公司; Re-52a型旋转蒸发器,上海亚荣生化仪器厂;APEX IIFT-ICR质谱仪。2 实验结果与讨论2.1 抑菌性能本次试验采用大肠杆菌和金黄色葡萄糖球菌,对罗布麻纤维、提取后的罗布麻纤维做了抑菌性能的对比试验,实验结果如图
6、1.1、1.2所示:棉纤维 罗布麻纤维 提取后的罗布麻纤维图1.1 培养4h的大肠杆菌专业好文档棉纤维 罗布麻纤维 提取后的罗布麻纤维图1.2 培养4h的金黄色葡萄糖球菌从两个图中我们可以明显的看到罗布麻纤维菌悬液中的菌落数明显少于棉的,这说明罗布麻纤维对大肠杆菌和金黄色葡萄糖球菌都具有不同程度的抑制性。我们用甲醇、乙醇、乙醇和水的混合物对罗布麻纤维中的有效成分进行提取后,纤维相对于没有经过提取的罗布麻纤维,菌落数明显增多,各种纤维的抑菌率如表1所示。从抑菌率我们可以明显的看出提取的罗布麻纤维的抑菌性能下降了很多,也就是说罗布麻纤维中的有效成分起到了抑菌作用。表1 几种纤维的抑菌率罗布麻 提取
7、罗布麻大肠杆菌 57% 27%金黄色葡萄糖球菌 34% 21.7%2.2 罗布麻纤维中黄酮类化合物的质谱分析现在普遍认为罗布麻纤维的抑菌性主要是由于其所含有的黄酮类化合物所致,所以我们就从黄酮类化合物入手,对罗布麻麻皮和罗布麻纤维中的黄酮类化合物进行了分析,结果如图 1.3 和 1.4 所示。对图 1.3 中的 A 和 B 进行比较,两个样品的质谱图存在明显的不同。麻皮质谱图中的比较明显的特征峰在罗布麻纤维中都已经消失,并且纤维质谱图中出现了许多新的特征峰。这说明罗布麻麻皮经过脱胶以后其所含有的中药成分都发生了明显的变化。A 图中有三个比较明显的负离子质谱特征峰,这三个负离子质谱特征峰的质荷比
8、即M-H -分别为 377、463、625,由于 z=1,所以该峰的质荷比就是该分子的分子量,经初步推测认为这三种分子可能为黄酮类化合物。为了进一步确定其成分,我们对原麻中的三个分子离子再轰击,使其进一步的裂解,得到二级质谱图,图 1.4,1.5。通过分析裂解碎片的成分,进一步确定原来的负离子质谱特征峰所对应的物质。专业好文档A 罗布麻麻皮提取液B 罗布麻纤维提取液图 1.3 质谱图专业好文档图 1.4 M-H-=377 负离子质谱特征峰的二级质谱图如图 1.4 所示,质荷比为 377 的负离子质谱特征峰被轰击裂解成质荷比为341 和 179 的两个离子碎片峰,M-H -为 179 的特征峰显
9、然是葡萄糖(C 6H12O6) ,它的分子量为 180;而M-H -为 341 的特征峰则对应双糖(C 12H22O12) ,它的分子量为 342.所以质荷比为 377 的负离子质谱特征峰所对应的物质是糖类,并非黄酮类化合物。A M-H-=463 负离子质谱特征峰专业好文档B M-H-=625 负离子质谱特征峰图 1.5 二级质谱图如图 1.5 A 所示,质荷比为 463 的负离子质谱特征峰被轰击裂解成质荷比为 301 的离子碎片峰,在前人研究的基础上,罗布麻纤维中含有的黄酮类化合物主要为槲皮素(C 15H10O7) ,计算其分子量为 302,与该碎片峰的M-H -相吻合,所以该峰所对应的物质
10、应该是五羟基黄酮即槲皮素。如图 1.5 B 所示,质荷比为 625 的负离子质谱特征峰被轰击后所形成的碎片峰与图 1.5A 中的相同,也就是说质荷比为 625 的分子离子峰对应的物质也是槲皮素,其比前者的质荷比多了 162,恰是 9 个 H2O 分子。所以,通过罗布麻麻皮的质谱图分析,原麻中的黄酮类化合物是质荷比为 463、625 的负离子质谱特征峰所对应的槲皮素。从图 1.5 中我们可以看到,罗布麻麻皮中含有的黄酮类化合物所对应的质谱特征峰在罗布麻纤维提取物的质谱图中并不存在,也就是说罗布麻纤维中实际上并不含有黄酮类化合物。这说明麻皮经过一系列的脱胶处理后,黄酮类化合物已经消失。麻皮中的黄酮
11、类化合物可能存在于木质素、果胶当中,在脱胶过程中,随着木质素和果胶一同被去除,也有可能罗布麻纤维中含有黄酮类化合物,但是在酸碱处理过程中被破坏。所以罗布麻纤维的抑菌性能与黄酮类化合物是无关的。在罗布麻纤维提取液的质谱图中也出现了许多新的特征峰,这些特征峰所对应的物质可能是中药成分经过一系列碱处理过程后发生变化后生成的新物质,也可能是甾醇、鞣质等其他中药成分,例如荷质比M-H -为 431的峰,推测可能为 -谷甾醇(C 29H50O),其分子量为 414,差值为 18 恰为一个水分子。正是这些物质的存在使得罗布麻纤维具有抑菌性。专业好文档3 结论罗布麻麻皮经过脱胶后,其所含有的黄酮类化合物已经消
12、失,所以推测罗布麻麻皮中的有效成分主要存在于果胶或木质素中,在纤维的脱胶过程中,果胶和木质素大多被去除,所以纤维中的有效成分明显减少。再者,在脱胶过程中,有效成分也可能被破坏掉。但是经过提取后的罗布麻纤维抑菌率明显降低,这说明罗布麻纤维中仍含有抑菌成分,这些有效成分可能是鞣质、- 谷甾醇等其他中药成分。参考文献:1 江佩芬.罗布麻叶脂溶性化学成分的研究中药通报,1985,122 陈奇云比色法测定罗布麻制剂中黄酮类化合物的含量中成药研究。1985,63 邢声远.天然医疗保健纤维.北京纺织,2001,3.4 卫生部消毒技术规范北京:中华人民共和创卫生部,19995 王俊起,王有斌纺织品抗菌功能方法
13、研究中国卫生工程学,2003,2专业好文档Editors note: Judson Jones is a meteorologist, journalist and photographer. He has freelanced with CNN for four years, covering severe weather from tornadoes to typhoons. Follow him on Twitter: jnjonesjr (CNN) - I will always wonder what it was like to huddle around a shortwave
14、radio and through the crackling static from space hear the faint beeps of the worlds first satellite - Sputnik. I also missed watching Neil Armstrong step foot on the moon and the first space shuttle take off for the stars. Those events were way before my time.As a kid, I was fascinated with what go
15、es on in the sky, and when NASA pulled the plug on the shuttle program I was heartbroken. Yet the privatized space race has renewed my childhood dreams to reach for the stars.As a meteorologist, Ive still seen many important weather and space events, but right now, if you were sitting next to me, yo
16、ud hear my foot tapping rapidly under my desk. Im anxious for the next one: a space capsule hanging from a crane in the New Mexico desert.Its like the set for a George Lucas movie floating to the edge of space.You and I will have the chance to watch a man take a leap into an unimaginable free fall f
17、rom the edge of space - live.The (lack of) air up there Watch man jump from 96,000 feet Tuesday, I sat at work glued to the live stream of the Red Bull Stratos Mission. I watched the balloons positioned at different altitudes in the sky to test the winds, knowing that if they would just line up in a
18、 vertical straight line “we“ would be go for launch.I feel this mission was created for me because I am also a journalist and a photographer, but above all I live for taking a leap of faith - the feeling of pushing the envelope into uncharted territory.The guy who is going to do this, Felix Baumgart
19、ner, must have that same feeling, at a level I will never reach. However, it did not stop me from feeling his pain when a gust of swirling wind kicked up and twisted the partially filled balloon that would take him to the upper end of our atmosphere. As soon as the 40-acre balloon, with skin no thic
20、ker than a dry cleaning bag, scraped the ground I knew it was over.专业好文档How claustrophobia almost grounded supersonic skydiverWith each twist, you could see the wrinkles of disappointment on the face of the current record holder and “capcom“ (capsule communications), Col. Joe Kittinger. He hung his
21、head low in mission control as he told Baumgartner the disappointing news: Mission aborted.The supersonic descent could happen as early as Sunday.The weather plays an important role in this mission. Starting at the ground, conditions have to be very calm - winds less than 2 mph, with no precipitatio
22、n or humidity and limited cloud cover. The balloon, with capsule attached, will move through the lower level of the atmosphere (the troposphere) where our day-to-day weather lives. It will climb higher than the tip of Mount Everest (5.5 miles/8.85 kilometers), drifting even higher than the cruising
23、altitude of commercial airliners (5.6 miles/9.17 kilometers) and into the stratosphere. As he crosses the boundary layer (called the tropopause), he can expect a lot of turbulence.The balloon will slowly drift to the edge of space at 120,000 feet (22.7 miles/36.53 kilometers). Here, “Fearless Felix“
24、 will unclip. He will roll back the door.Then, I would assume, he will slowly step out onto something resembling an Olympic diving platform.Below, the Earth becomes the concrete bottom of a swimming pool that he wants to land on, but not too hard. Still, hell be traveling fast, so despite the distan
25、ce, it will not be like diving into the deep end of a pool. It will be like he is diving into the shallow end.Skydiver preps for the big jumpWhen he jumps, he is expected to reach the speed of sound - 690 mph (1,110 kph) - in less than 40 seconds. Like hitting the top of the water, he will begin to
26、slow as he approaches the more dense air closer to Earth. But this will not be enough to stop him completely.If he goes too fast or spins out of control, he has a stabilization parachute that can be deployed to slow him down. His team hopes its not needed. Instead, he plans to deploy his 270-square-
27、foot (25-square-meter) main chute at an altitude of around 5,000 feet (1,524 meters).In order to deploy this chute successfully, he will have to slow to 172 mph (277 kph). He will have a reserve parachute that will open automatically if he loses consciousness at mach speeds.Even if everything goes a
28、s planned, it wont. Baumgartner still will free fall at a speed that would cause you and me to pass out, and no parachute is guaranteed to work higher than 25,000 feet (7,620 meters).It might not be the moon, but Kittinger free fell from 102,800 feet in 1960 - at the dawn of an infamous space race t
29、hat captured the hearts of many. Baumgartner will attempt to break that record, a feat that boggles the mind. This is one of those monumental moments I will always remember, because there is no way Id miss this.专业好文档WXmA&UE9aQGn8xp$R#͑GxGjqv$UE9wEwZ#QcUE%&qYpEh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3
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33、NGpP$vSTT#&ksv*3tnGK8!z9JWKffwvG#tYM*Jg&6a*CZ7H$dq8KqqfHVZFedswSyXTy#&QA9wkxFyeQ!djs#XuyUP2kNXpR89AmYWpazadNu#KN&MuWFA5uxY7JnD6YWRrWwcvR9CpbK!zn%Mz849GxG89AmUE9aQGn8xp$R#͑GxGjqv$UE9wEwZ#QcUE%&qYpEh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu#KN&MuWFA5uxY7JnD6YWRrWwcvR9CpbK!zn%Mz849GxG
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35、KqqfHVZFedswSyXTy#&QA9wkxFyeQ!djs#XuyUP2kNXpRWXmA&UE9aQGn8xp$R#͑GxGjqv$UE9wEwZ#QcUE%&qYpEh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu#KN&MuWFA5uxY7JnD6YWRrWwcvR9CpbK!zn%Mz849GxGjqv$UE9wEwZ#QcUE%&qYpEh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu#KN&MuWFA5uxGjqv$UE9wEwZ#QcUE
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37、R9CpbK!zn%Mz849GxGjqv$UE9wEwZ#QcUE%&qYpEh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu#KN&MuWFA5uxGjqv$UE9wEwZ#QcUE%&qYpEh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu#KN&MuWFA5uxY7JnD6YWRrWwcvR9CpbK!zn%Mz849GxGjqv$UE9wEwZ#QcUE%&qYpEh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89Am
38、YWv*3tnGK8!z89AmYWpazadNu#KN&MuWFA5uxY7JnD6YWRrWwcvR9CpbK!zn%Mz849GxGjqv$UE9wEwZ#QcUE%&qYpEh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNuGK8!z89AmYWpazadNu#KN&MuWFA5uxY7JnD6YWRrWwcvR9CpbK!zn%Mz849GxGjqv$UE9wEwZ#QcUE%&qYpEh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu#KN&MuWFA5uxG
39、jqv$UE9wEwZ#QcUE%&qYpEh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWpazadNu#KN&MuWFA5uxY7JnD6YWRrWwcvR9CpbK!zn%Mz849GxGjqv$UE9wEwZ#QcUE%&qYpEh5pDx2zVkum&gTXRm6X4NGpP$vSTT#&ksv*3tnGK8!z89AmYWv*3tnGK8!z89AmYWpazadNu#KN&MuWFA5uxY7JnD6YWRrWwcvR9CpbK!zn%Mz849GxGjqv$U*3tnGK8!z89AmYWpazadNu#KN&MuWFA5uxY
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