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1.
为了了解菊黄东方鲀(Takifugu flavidus)、暗纹东方鲀(T.obscurus)及其杂交F1代的肌肉营养特征,利用生物化学方法,从每类实验样本中取9尾对其肌肉中的粗蛋白、粗脂肪、水分、粗灰分和氨基酸成分进行了测定和分析。结果显示:(1)杂交F1代在生长方面具有明显的杂交优势,与亲本之间存在着显著差异(P0.05),杂交F1代的体重为其亲本的1.48~1.77倍;(2)杂交F1代肌肉水分含量与其母本含量相近,但粗脂肪含量均较亲本少(P0.05),粗蛋白含量则与亲本差异不显著(P0.05);(3)除色氨酸和胱氨酸外,16种氨基酸均在肌肉样本中被检测到,除甲硫氨酸外,其余15种氨基酸间含量均存在着显著性差异(P0.05)。菊黄东方鲀(♀)×暗纹东方鲀(♂)杂交F1代的总氨基酸含量最高,而暗纹东方鲀(♀)×菊黄东方鲀(♂)F1代总氨基酸含量则介于两亲本之间。对其必需氨基酸总量进行分析发现,菊黄东方鲀与其正反杂交F1代之间均存在显著性差异(P0.05),而暗纹东方鲀与其正反杂交F1代之间差异不显著(P0.05);(4)肌肉营养品质评价结果表明,菊黄东方鲀(♀)×暗纹东方鲀(♂)杂交F1代的鲜味氨基酸含量为26.68%,明显高于双亲样本(菊黄东方鲀22.28%、暗纹东方鲀25.20%),而暗纹东方鲀(♀)×菊黄东方鲀(♂)F1代的鲜味氨基酸总量(23.30%)较其父本偏高,但低于其母本。研究结果表明,杂交东方鲀的肌肉营养综合了双亲的优良特性,特别是是菊黄东方鲀(♀)×暗纹东方鲀(♂)杂交F1代,拥有最高的鲜味氨基酸含量,具有推广价值。  相似文献   

2.
在基础饲料中分别添加不同浓度的益生菌(0.1%、0.2%、0.4%)、壳聚糖(0.2%、0.5%、1.0%)、壳聚糖与益生菌混合物、甘露聚糖与益生菌混合物,在室内水泥池(5.0m×2.0m×1.0m)中喂养暗纹东方鲀幼鱼(3.15±0.05g),用基础饲料投喂暗纹东方鲀作为对照,每组三个重复,60d后测定鱼体的生长、肌肉生化成分、肝胰脏、肠道蛋白酶和淀粉酶活性和白肌RNA/DNA。结果显示,对照组相对增重率为476.4%,饲料中添加0.2%的壳聚糖、0.1%的益生菌对暗纹东方鲀的促生长作用最明显,相对增重率显著提高。壳聚糖与益生菌混合物、甘露聚糖与益生菌混合物对生长无显著影响。对照组肌肉蛋白质和脂肪含量分别为82.94%、11.21%,壳聚糖、益生菌与甘露聚糖在饲料中的添加,能够显著降低肌肉脂肪含量,显著提高肌肉蛋白质含量,也能在一定程度上降低肝体比。0.2%壳聚糖和0.1%益生菌能使肠道淀粉酶活性显著增强(P<0.05),白肌RNA/DNA显著增大(P<0.05),说明肠道淀粉酶活性增强和蛋白质合成增加是壳聚糖和益生菌促进暗纹东方鲀生长的重要原因。  相似文献   

3.
为探讨菊粉对暗纹东方鲀(Takifugu obscures)幼鱼生长、消化及免疫能力的影响,利用单因素实验设计,选取640尾暗纹东方鲀,体重(6.97 ± 1.32)g,随机分为4组,每组设4个重复,在日粮中添加不同浓度的菊粉(0%、0.25%、0.5%、1%),连续投喂8周后测定鱼体生长、肌肉营养成分、肠道消化酶和肝非特异性免疫酶活性。结果显示,各菊粉添加组的生长指标无显著性差异(P > 0.05);0.25%组肌肉的粗脂肪含量(0.87%)显著高于0.5%组(0.82%)和1%组(0.76%);0.25%组肌肉的粗蛋白含量(18.75%)显著高于1%组(18.50%),与对照组(18.62%)和0.5%组(18.60%)无显著差异。暗纹东方鲀幼鱼肠道组织的胃蛋白酶、胰蛋白酶、脂肪酶和淀粉酶活性均随着菊粉添加量的增加而呈逐渐上升的趋势。0.5%菊粉添加组肝组织的谷胱甘肽过氧化物酶和总超氧化物歧化酶活性极显著(P < 0.01)高于其余处理组;0.5%组的过氧化氢酶活性也极显著(P < 0.01)高于0.25%和1%组;丙二醛含量随着菊粉添加量的增加而呈递减的趋势。因此,菊粉添加量以0.5%为宜,对暗纹东方鲀幼鱼的生长无显著影响,但对其消化酶活性和非特异性免疫酶活性均具有促进作用。  相似文献   

4.
本实验采用薄层色谱法对川楝子中香草酸、异香草酸进行定性鉴别;采用高效液相色谱法对川楝子中香草酸进行含量测定。色谱柱:Hypersil BDS C18(250 mm×4.60 mm,5μm),柱温:30℃,流动相:甲醇∶水∶冰乙酸=25∶75∶0.5,流速:1.0 mL.min-1,检测波长:260 nm。最终,定性鉴别斑点清晰。结果表明香草酸含量在1.022~16.352μg范围内,进样量与峰面积呈良好线性关系,相关系数r=0.9998;香草酸的平均回收率为102.45%,RSD=1.22%(n=6)。  相似文献   

5.
暗纹东方鲀线粒体COI及其侧翼tRNA基因的克隆与序列分析   总被引:8,自引:0,他引:8  
邵爱华  朱江  陈葵  史全良  姚炜雯 《遗传》2006,28(8):963-971
以暗纹东方鲀(Takifugu fasciatus)肝脏的线粒体DNA为模板,按照红鳍东方鲀线粒体DNA序列设计合成特异引物进行PCR扩增,克隆并测定了线粒体细胞色素氧化酶I亚基(COI)及其侧翼tRNA基因的全序列,结果显示,克隆了暗纹东方鲀COI基因1546bp及其5′端上游的tRNATyr基因和3′端下游的tRNASer基因序列共1766bp。用DNA分析软件对暗纹东方鲀与GenBank中10个目13种鱼类的COI序列进行比较分析,显示暗纹东方鲀与这些鱼类的COI基因具有较高的同源性,与同属红鳍东方鲀的同源性最高为97.6%,与同目不同科的矛尾翻车鲀和翻车鲀的同源性为76.5%和75.4%。根据暗纹东方鲀与其他13种鱼的COI基因序列同源性所建立的进化树,与传统的分类地位基本吻合。推定的这二种tRNA的二级结构都具有典型的三叶草型结构。  相似文献   

6.
林子安  昌水平  刘庭恩 《蛇志》2014,(2):153-155
目的研究拔毒消炎软膏的质量控制方法。方法用薄层色谱法(HPLC)对制剂中大黄、黄柏进行定性鉴别,高效液相色谱法测定大黄酚的含量。结果定性鉴别薄层色谱斑点特征明显;高效液相色谱法测定含量,大黄酚在0.061~0.304μg/ml范围内呈良好的线性关系(r=0.9954),平均加样回收率为98.68%,RSD=1.39%。结论采用薄层色谱法定性、高效液相色谱法定量,简便准确、重现性良好,可有效控制该制剂质量。  相似文献   

7.
为了解雌激素在鱼类雌性性别分化中的作用,在克隆暗纹东方鲀(Takifugu obscurus)性腺CYP19A和DMRT1基因部分序列基础上,采用不同浓度芳香化酶抑制剂来曲唑(letrozole,LE)(0、25、125、625、3 125μg/L)处理暗纹东方鲀初孵仔鱼,每个平行组各20尾鱼,观察CYP19A和DMRT1基因的表达情况和组织学变化。RT-PCR结果显示:25μg/L LE实验组中样本CYP19A和DMRT1表达水平与对照组相比无显著差异;LE 125μg/L以上各浓度组中约30%的样本同时表达CYP19A和DMRT1基因,且随着LE浓度增加,CYP19A表达量下调,而DMRT1表达量上调;组织学研究表明,125μg/L LE以上各实验组样本中可见由雌性向雄性转变的间性体性腺。孵化后56 d,125μg/L LE实验组样本中约有20%显示雌性;625μg/L LE实验组样本显示表型雄性和间性体,未见雌性;最高浓度3 125μg/L LE组中所有样本均显示表型雄性,CYP19A和DMRT1基因表达与对照组雄性相同。上述结果说明,抑制内源雌激素合成可使暗纹东方鲀仔稚鱼CYP19A基因表达下调,同时DMRT1基因表达上调,并发生雄性化性逆转。  相似文献   

8.
反相高效液相色谱法检测脂必妥片中洛伐他汀的含量   总被引:1,自引:0,他引:1  
对脂必妥片样品进行预处理,得到的混合物用薄层色谱、反相高效液相色谱法进行定性、定量分析,建立检测其含量的方法。当色谱条件为色谱柱:Kromasil(C18250 mm×4.6 mm,5μm);流动相体积比:乙腈∶水为85∶15,;流速:1.0 ml.min-1;检测波长:420 nm;柱温:30℃;测定结果表明被测峰和其它峰可完全分离,在每毫升10.21~200.03μg内具有良好的线性关系,r=0.9997,测得其中洛伐他汀含量为0.25%,回收率:97.73%,RSD:0.56%。这种方法准确、可靠,可用于含洛伐他汀药品的质量控制。  相似文献   

9.
长江的鲀形目鱼类资源主要有两种,暗纹东方鲀(Fugu obscurus Abe)和弓斑东方鲀Fugu ocellatus (Linnaeus).研究结果表明:与野生的弓斑东方鲀相比较,野生的暗纹东方鲀的毒性比较低.现在已有人工养殖的暗纹东方鲀,那么它们的毒性是否也像红鳍东方鲀一样,经过人工养殖后毒性有所下降,甚至变为无毒,因此食用更加安全了呢?为此,作者进行了研究.  相似文献   

10.
建立了自动在线柱前衍生反相高效液相色谱法同时测定γ-氨基丁酸(GABA)和17种游离氨基酸含量的方法.以邻苯二甲醛-9-芴基甲基氯甲酸酯(OPA-FMOC)为衍生试剂进行衍生,Agilent Hypersil AA-ODS-C18色谱柱分离,梯度洗脱,二极管阵列检测器检测,在19min内分离测定了马尾松苗木针叶中GABA 和17种游离氨基酸的含量.该方法测定氨基酸的回收率高于90.1%,精密度和重现性均较好(相对标准偏差为0.21%~2.81%),经测定,发现马尾松被马尾松毛虫取食后,所测18种氨基酸总量明显降低,从418.3μg · g-1降低到310.4μg · g-1鲜重.  相似文献   

11.
We studied the role of induced plant phenols as a defense response to insect herbivory. Phenolic compounds were induced in Capsicum annuum L., the source of many culinary peppers, after feeding by different stages of the insect pest, Spodoptera litura F. The phenols were identified and quantified using high performance liquid chromatography (HPLC) and effects produced by these phenols on larval development were studied. Vanillic acid was identified in plants challenged by second, fourth, and fifth instar larvae, but not in plants challenged by third instar nor unchallenged plants. Syringic acid production was induced in chili plants infested with second (0.429 ± 0.003 μg/g fresh weight, fourth (0.396 ± 0.01 μg/g fresh weight), and fifth instar (5.5 ± 0.06 μg/g fresh weight) larvae, compared to untreated plants (0.303 ± 0.01 μg/g fresh weight) plants. Leaves surface treated with the rutin deterred oviposition. Dietary exposure to chlorogenic acid, vanillic acid, syringic acid, sinapic acid, and rutin led to enhanced activities of detoxifying enzymes, β‐glucosidase, carboxyl esterase, glutathione S‐transferase, and glutathione reductase in the midgut tissues of all the larval instars, indicating the toxic nature of these compounds. Protein carbonyl content and acetylcholinesterase activity was analyzed to appreciate the role of induced plant phenols in insect protein oxidation and terminating nerve impulses.  相似文献   

12.
采用高效液相色谱法对不同时期、不同品种的梨花中熊果苷的含量进行了分析,色谱柱为Hypersil BDSC18(4.6 mm×250 mm,5μm),流动相为甲醇∶水(6∶94),加入甲酸0.05%,检测波长280 nm。结果表明:熊果苷在0.01~5.00μg范围内线性关系良好(r=0.9999),平均加标回收率为97.7%。不同生长期的鸭梨梨花中熊果苷的含量一般在10 mg/g以上,尤以花芽萌动期时含量最高,达到35.7 mg/g(鲜重计)。不同品种的9份梨花样品中,熊果苷含量在3.5~10.5 mg/g之间。  相似文献   

13.
Proportions between oxidized and reduced glutathione forms were determined in vacuoles isolated from red beet (Beta vulgaris L.) taproots. The pool of vacuolar glutathione was compared with glutathione pools in isolated plastids and mitochondria. The ratio of glutathione forms was assessed by approved methods, such as fluorescence microscopy with the fluorescent probe monochlorobimane (MCB), high-performance liquid chromatography (HPLC), and spectrophotometry with 5,5′-dithiobis-2-nitrobenzoic acid (DTNB). The fluorescence microscopy revealed comparatively low concentrations of reduced glutathione (GSH) in vacuoles. The GSH content was 104 μM on average, which was lower than the GSH levels in mitochondria (448 μM) and plastids (379 μM). The content of reduced (GSH) and oxidized (GSSG) glutathione forms was quantified by means of HPLC and spectrophotometric assays with DTNB. The glutathione concentrations determined by HPLC in the vacuoles were 182 nmol GSH and 25 nmol GSSG per milligram protein. The respective concentrations of GSH and GSSG in the plastids were 112 and 6 nmol/mg protein and they were 228 and 10 nmol/mg protein in the mitochondria. The levels of GSH determined with DTNB were 1.5 times lower, whereas the amounts of GSSG were, by contrast, 1.5–2 times higher than in the HPLC assays. Although the glutathione redox ratios depended to some extent on the method used, the GSH/GSSG ratios were always lower for vacuoles than for plastids and mitochondria. In vacuoles, the pool of oxidized glutathione was higher than in other organelles.  相似文献   

14.
目的建立心痛宁滴丸中香附的薄层色谱鉴别和阿魏酸的分析方法。方法用TLC法鉴别香附中的α-香附酮;用HPLC法测定川芎中阿魏酸的含量,Hypersil ODS-C18分析柱(250 mm×4.6 mm,5μm)检测,波长:323 nm,流动相:甲醇-水-冰醋酸(30∶70∶0.7),流速:l ml/min,柱温:室温。结果薄层法鉴别香附中的香附酮结果满意;HPLC法测阿魏酸,线性范围0.005-0.5μg(r=0.9999),回收率分别为98.0%(n=6)。结论上述方法准确、专属性强,可作为心痛宁滴丸的香附定性和阿魏酸定量的分析方法。  相似文献   

15.
5-(Pentafluorobenzoylamino)fluorescein (PFB-F), a new thiol-reactive molecule was synthesized to improve the detection limits and specificity of the assays for glutathione S-transferase (GST) activity and glutathione (GSH). A rapid assay method to measure GSH concentration or GST activity and the simultaneous analysis of multiple samples is possible because the glutathione adduct, GS-TFB-F, is separated from PFB-F by thin-layer chromatography (TLC) and can be quantitated by a fluorescence scanner. The detection limits for GSH and for GST activity using TLC were found to be as low as 10 pmol/microl and 1 ng/microl using equine liver GST, respectively. Determination of GSH concentration or GST activity in bovine pulmonary artery endothelial (BPAE) cell lysates gave a linear response for samples corresponding to 500-2500 cells. PFB-F could also measure GST activities of GST fusion proteins and prove to be a suitable substrate for determining the activities of human GST isozymes and other sources of mammalian GST. The selectivity of PFB-F with GSH was proven by comparing trace amount of the adducts that formed with cysteine and beta-galactosidase to that formed with GSH. The HPLC profile of a reaction mixture where cell lysate was used in place of purified GST, also shows only two main peaks, corresponding to GS-TFB-F and unreacted PFB-F. The selectivity of PFB-F for GSH was further confirmed by exposing BPAE cells to dl-buthionine-[S,R]-sulfoximine (BSO). Our results of GS-TFB-F determination indicate that 12-, 24-, or 36-h incubations with BSO caused 2-, 6-, or 7.6-fold reductions in GSH levels, respectively.  相似文献   

16.
A monoclonal antibody produced to abscisic acid (ABA) has been characterised and the development of a radioimmunoassay (RIA) for ABA using the antibody is described. The antibody had a high selectivity for the free acid of (S)-cis, trans-ABA. Using the antibody, ABA could be assayed reliably in the RIA over a range from 100 to 4000 pg (0.4 to 15 pmol) ABA per assay vial. As methanol and acetone affected ABA-antibody binding, water was used to extract ABA from leaves. Water was as effective as aqueous methanol and acetone in extracting the ABA present. Crude aqueous extracts of wheat, maize and lupin leaves could be analysed without serious interference from other immunoreactive material. This was shown by measuring the distribution of immunoreactivity in crude extracts separated by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC), or by comparing the assay with physicochemical methods of analysis. Analysis of crude extracts by RIA and either, after TLC purification, by gas chromatography using an electron-capture detector or, after HPLC purification, by combined gas chromatography-mass spectrometry (GC-MS) gave very similar ABA concentrations in the initial leaf samples. However, RIA analysis of crude aqueous extracts of pea seeds resulted in considerable overestimation of the amount of ABA present. Determinations of ABA content by GC-MS and RIA were similar after pea seed extracts had been purified by HPLC. Although the RIA could not be used to analyse ABA in crude extracts of pea seeds, it is likely that crude extracts of leaves of several other species may be assayed successfully.Abbreviations ABA abscisic acid - DW dry weight - FW fresh weight - GC-ECD gas chromatography using an electron capture detector - GC-MS combined gas chromatographymass spectrometry - HPLC high-performance liquid chromatography - McAb monoclonal antibody - PVP soluble polyvinylpyrrolidone - RIA radioimmunoassay - TLC thin-layer chromatography  相似文献   

17.
为选择一种准确快捷的方法测定银耳多糖的单糖组成,对薄层色谱法(TLC)、气相色谱法(GC)、高效液相色谱法(HPLC)三种色谱方法进行比较。结果表明,前两种方法的测定结果均不理想,而HPLC法,操作简便,灵敏度高,分离效果好,信息完整。测定结果为由葡萄糖、甘露糖、葡萄糖醛酸、木糖、岩藻糖组成,其摩尔比为0.24∶1.00∶0.06∶0.29∶0.25。HPLC法对酸性杂多糖组成糖分析是一种比较理想的选择。  相似文献   

18.
para-Sulfonylbenzoyloxybromobimane (sBBr) was shown to be similar to the fluorescent labeling agent monobromobimane (mBBr) in reacting rapidly and selectively with thiols to produce stable derivatives which are readily separated by HPLC. Chromatography of the sBBr derivative provides a useful means of confirming the identification of an unknown thiol based upon the chromatography of its mBBr derivative and can be useful for quantitative determination of polycationic thiols for which chromatography of the mBBr derivative is unsatisfactory. Unlike mBBr, which readily penetrates cells, sBBr was found not to be taken up by cells. These characteristics allow sBBr to be used, in conjunction with mBBr, to quantify the export of thiols from cells, as illustrated for GSH and the radioprotective drug WR1065, from V79 cells. Simultaneous determination of GSH and glutathione disulfides in cell culture medium could be achieved by labeling of thiols with sBBr followed by reduction of disulfides with dithiothreitol, labeling of the resulting thiols with mBBr, and HPLC analysis for both glutathione derivatives.  相似文献   

19.
Winter wheat (Triticum sativum L. ev. Nisu) was grown in sand which contained 0, 0.25, 0.5 and 1.0 mg S-ethyl dipropylthicarbamate (EPTC) per kg air dry sand. In 21 day old roots, the phospholipids (PL) were extracted in ice-cold chloroform/methanol (2:1, v/v) and isolated by thin layer chromatography (TLC). The PL fatty acids were analysed by gas liquid chromatography (GLC). The major fatty acids of the root PL fraction were palmitic, oletic, linoleic, and linolenic (22.4, 6.8, 39.2, and 23.1 μ/g root fresh weight, respectively). Total fatty acid content of the PL fraction was decreased to 39% by 1 mg EPTC/kg sand in which linolenic acid was decreased to 28%. The remainder of the major fatty acid constituents were decreased to 12–47%. The general quality of fatty acids in the PL fraction was slightly altered, while a 60% inhibition of total PL production resulted.  相似文献   

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