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两种固相萃取柱在乙酰氨基阿维菌素药动学前处理中的比较 总被引：1，自引：0，他引：1
为评价乙酰氨基阿维菌素(EPR)药动学前处理中两种固相萃取(SPE)柱的净化、萃取效果,首先在空白胎牛血清中加入一定量的EPR标准溶液,经色谱甲醇提取后,分别经Agilent和Waters公司的C_(18) SPE柱对其进行萃取,再经N-甲基咪唑和三氟乙酸酐衍生化,最后用超高效液相色谱-荧光检测器进行测定;同时,将等量的EPR标准溶液进行衍生化,用超高效液相色谱-荧光检测器进行测定,将其结果作为对照;以绝对回收率作为评价两种SPE柱净化、萃取EPR效果的标准。结果显示,含有EPR的胎牛血清分别经Agilent和Waters公司的C_(18) SPE柱萃取后,两者的EPR峰形均尖锐对称且无明显杂质峰,但绝对回收率有差异,分别为94.50%和85.79%。结果表明,Agilent公司的C_(18) SPE柱对EPR的萃取效果更理想。 相似文献
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初步评价在乙酰氨基阿维菌素药动学前处理中3种固相萃取柱对乙酰氨基阿维菌素的净化、萃取效果。首先在空白胎牛血清中加入一定量的乙酰氨基阿维菌素标准溶液,经色谱甲醇提取后,分别经Agilent公司、Waters公司和Agela Technologies公司的C18SPE柱进行萃取(分别简称A固相萃取柱、W固相萃取柱、AT固相萃取柱),再经N-甲基咪唑和三氟乙酸酐衍生化,最后用超高效液相色谱-荧光检测器进行测定。与此同时,将等量的乙酰氨基阿维菌素标准溶液进行衍生化,用超高效液相色谱-荧光检测器进行测定,其结果作为对照。以绝对回收率作为评价3种SPE柱净化、萃取EPR效果的标准。结果显示,含有EPR的胎牛血清经A固相萃取柱萃取后,EPR峰形尖锐对称且无明显杂质峰,其绝对回收率为80.79%;经W固相萃取柱萃取后,乙酰氨基阿维菌素峰形尖锐对称且无明显杂质峰,其绝对回收率为83.61%;经AT固相萃取柱萃取后,乙酰氨基阿维菌素峰形尖锐对称且无明显杂质峰,其绝对回收率仅为64.20%。相对于A与AT固相萃取柱,Waters公司的C18SPE柱萃取EPR的效果更好。研究结果为后期乙酰氨基阿维菌素的药物代谢动力学试验奠定了基础。 相似文献
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采用高效液相色谱法（HPLC），分离柱选用ODS色谱柱，以纯水为洗脱液，用示差折光检测。测定了饲料添加剂和功能食品中低聚糖的含量，即蔗糖、水苏糖和棉子糖。本法具有流动相价廉、无污染、快速简便和稳定性好的特点，色谱柱使用寿命长。 相似文献
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UPLC和HPLC法检测猪肉中磺胺类药物残留的比较 总被引：1，自引：0，他引：1
比较了超高效液相色谱法与高效液相色谱法测定猪肉中磺胺类药物残留的结果。采用农牧发[2001]38号动物源食品中磺胺类药物残留的检测方法有关标准,将原高效液相色谱方法转换并优化为超高效液相色谱方法。高效液相色谱方法色谱柱为Waters Symmetry C18柱(4.6 mm×250 mm,5μm),流动相:乙腈-甲醇-水-乙酸(2∶2∶9∶0.2),流速:1 mL/min,进样量:50μL;超高效液相色谱方法色谱柱为Waters Acquity UPLC BEH RP18柱(2.1 mm×50 mm,1.7μm),流动相:乙腈-甲醇-水-乙酸(2∶2∶10∶0.2),流速:0.4 mL/min,进样量:5μL。结果表明超高效液相色谱方法能够替代高效液相色谱分析方法测定猪肉中磺胺类药物残留,既加快了分析速度,达到了样品分析的高通量,减少有机溶剂的使用,又得到更高的分析灵敏度。 相似文献
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本文提供了同时测定牛奶中磺胺嘧啶(SDZ)、磺胺二甲基嘧啶(SMZ)、三甲氧苄二氨嘧啶(TMP)和对氨基苯甲酸(PABA)残留的快速、灵敏的高效液相色谱测定方法。样品的前处理非常简单,只需要脱蛋白和离心,没有费时繁琐的有机提取、相分离和蒸发浓缩等步骤。提取液在一反相μBondpak C_(18)径向加压柱上进行色谱分离,用254nm紫外检测。该方法测定范围宽,线性响应好,回收率高,值得推广使用。 相似文献
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Abomasal parasites inhibit gastric acid secretion and reduce the number of acid secreting parietal cells either through physical contact with gastric tissue, the release of inhibitory excretory/secretory (ES) products or by initiating the host inflammatory response. To examine the role of parasite ES products, adult Haemonchus contortus were incubated in a medium based on Hank's balanced salt solution and three incubates were tested for the presence of inhibitors of acid secretion by dispersed rabbit gastric glands in vitro, using the intracellular accumulation of 14C-aminopyrine (AP) as an indirect measure of acid secretion. Acceptably sensitive rabbit preparations (80%) for testing ES products showed increased 14C-AP accumulation with either 10(-5) or 10(-4)M histamine. Sheep glands proved unsuitable as a test system as only two of six preparations responded weakly to histamine. Inhibitory activity was demonstrated in all the three parasite incubates, although responses varied quantitatively between tests, even with the same batch of incubate. In single tests, 16% ES products significantly reduced the 14C-AP ratio (P<0.05) of 10(-4)M histamine-stimulated glands (15 of 19 tests with Batch 1 and two of six with Batch 3). Even at 1 and 5%, ES products (Batch 2) were inhibitory for 10(-5)M histamine-stimulated glands: at 1% the mean reduction was 26.0% (range 7.8-54.8%, n=10), four of which were statistically significant and at 5% was 17.6% (range -15.3-53.8%, n=17), four of which were statistically significant. ES products were overall inhibitory (-12%, P<0.05), when tested on glands exposed to increasing histamine concentrations from 10(-6)M to 10(-3)M (which increased the 14C-AP ratio (P<0.001)). Responses by individual gland preparations differed (P<0.001). The active component(s) of the parasite ES products were less than 5000 molecular weight. Ammonium chloride, tested over the range of concentrations of ammonia present in worm incubates (0.2-1 mM, final concentrations in the gland incubations of 0.005-0.1 mM), had variable, but overall inhibitory effects on 10(-5)M histamine-stimulated glands (n=3). When tested with a range of histamine concentrations, 0.01 mM NH4Cl overall reduced the response by 8.6% (P<0.05)(n=4). The similarity of responses of rabbit glands to parasite ES products and to NH4Cl suggests that ammonia may be the small molecular weight ES product of adult H. contortus, which inhibited acid secretion in vitro. 相似文献
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高效液相色谱法测定蜂蜜中β-呋喃果糖苷酶酶活 总被引：1，自引：0，他引：1
建立了一种检测蜂蜜中β-呋喃果糖苷酶酶活的方法，即样液中加棉籽糖作为反应底物，在pH为5．4的缓冲液中，45aC摇床200r／min条件下反应2h，测定样液中的密二糖含量。色谱条件为：Phenomenex NH2 Luan 5U 250×4．60mm；流动相：乙腈：水=80：20；进样量：10μL；流速：1．2mL／min；柱温：40℃；示差折光检测器，温度40cc。实验结果表明：β-呋喃果糖苷酶酶活在0．3—1．7U／mL范围内线性良好，相关系数R为0．998 10。相对标准偏差为1．72％（n=5），平均回收率为94．5％。该分析方法简便、准确、灵敏，重现性好，适用于蜂蜜中β-呋喃果糖苷酶酶活的检测。 相似文献
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试验旨在建立灵敏、快速、特异的氯霉素（CAP）检测方法,基于免疫亲和色谱技术和酶联免疫技术,研究了一种新型快速检测虾肉中CAP的免疫亲和色谱柱（ICTC）检测方法。采用溶胶-凝胶（sol-gel）法制备ICTC柱的检测层和控制层,优化检测层、控制层、酶标物（CAP-HRP）稀释度等条件,制备ICTC柱。测定ICTC柱的灵敏度、特异性等性能参数,并对虾肉样本前处理方法和基质干扰效应进行优化。本研究确定ICTC柱条件为：检测层,anti-CAP sol-gel：空白sol-gel为10：1 000（V/V）;控制层,anti-HRP sol-gel：空白sol-gel为13：1 000（V/V）;CAP-HRP的稀释度为1：105。采用检测层消色法判定结果,检测时间10 min,PBS缓冲液中CAP的检测限为0.5 μg/L,该方法与甲砜霉素、氟苯尼考、氟苯尼考胺同类药物或代谢产物均无可见交叉反应,在虾肉样本中的检测限为0.75 μg/kg。本研究建立的ICTC柱检测方法快速、灵敏,使用方便,为现场快速检测虾肉中CAP残留提供了新的技术手段。 相似文献
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受地形和气候的影响,高寒草地土壤经历着频繁的土壤水分波动过程,为探索土壤水分波动对青藏高原高寒草甸生态系统CO_2和N_2O排放的影响,采用原状土柱模拟土壤由高含水量(60 cm3·cm-3)到低含水量(30 cm3·cm-3)再到高含水量(60 cm3·cm-3)的波动过程,各阶段持续时间相应为38、57和46 d,并以恒定含水量(60 cm3·cm-3)为对照,研究了高寒草甸生态系统CO_2和N_2O的释放量及其与土壤温湿度、土壤铵态氮(NH4+-N)、硝态氮(NO3--N)和可溶性有机碳(DOC)的关系。结果表明,土壤水分波动增加了高寒草甸生态系统CO_2排放通量。在土壤水分波动过程中,CO_2排放通量与土壤温度呈正指数关系;在开始的高含水量阶段,CO_2排放与DOC含量有显著正相关关系(P0.05),但在低含水量阶段,相关性不明显(P0.05);土壤水分波动显著降低了N_2O的排放(P0.05)。恒湿对照的N_2O通量与其土壤NH4+-N含量呈极显著负相关关系(P0.01),土壤水分波动处理的N_2O通量与土壤NH4+-N和NO3--N含量相关性均不显著(P0.05)。 相似文献
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The objective of this study was to determine the agreement between ELISA tests conducted using three O. ostertagia antigens: crude adult worm, larval stage 4 (L4) excretory/secretory (ES) and adult ES. This study was carried out on 289 Holstein cows from five herds in Prince Edward Island and one herd in Nova Scotia. Composite milk samples of these cows were collected (between May and September 2002) from the respective provincial laboratories and sent to the Atlantic Veterinary College where each sample was tested for antibodies to O. Ostertagi using an indirect microtitre ELISA test. Results were expressed as optical density ratio (ODR) values. Each milk sample was tested with three ELISA tests, with each test using a different O. ostertagi antigen. There was a slight rise in ODR values of both adult antigens, between May and August, with higher values obtained using the adult ES antigen. L4 ES ODR values were generally higher than those for both adult antigens during the study period, except for May. There was a more dramatic rise in L4 ES ODR values between May and August. Rises in ODR in May and end of July coincided with periods of mass maturation of L4 to adult worms. The results of the study showed that the concordance correlation coefficient (CCC) between tests performed using both ES and the crude antigens were low (crude adult versus adult ES=0.31, crude adult versus L4 ES=0.30). The highest CCC was observed between tests done using both ES antigens (CCC=0.56). Generally, the study results suggest that the antibody response (detectable by the ELISA) is mainly directed against ES antigens (especially L4) than the crude adult worm antigen. 相似文献
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Qualitative and quantitative evaluation of coprological and serological techniques for the diagnosis of fasciolosis in cattle 总被引：1，自引：0，他引：1
Charlier J De Meulemeester L Claerebout E Williams D Vercruysse J 《Veterinary parasitology》2008,153(1-2):44-51
The objectives of this study were (1) to evaluate available coprological and serological tests for detection of Fasciola hepatica infection in field conditions, (2) to investigate if the season when samples were collected affects the interpretation of the test results, and (3) to evaluate if the test results are associated with the level of infection. During weekly visits to an abattoir, the whole liver, a rectal faecal sample and a blood sample were collected from 100 cows in two seasons each ("spring"=February-May 2006 and "autumn"=October-December 2006). A sedimentation-flotation technique on 4g (SF 4g) or 10g (SF 10g) of faeces, a copro-antigen ELISA and two indirect serum F. hepatica ELISAs (excretory-secretory (ES) and Pourquier ELISA) were performed and the test results were compared with the presence of infection and worm counts at liver necropsy. Over both seasons the sensitivity (Se) and specificity (Sp) were for the SF 4g 43% and 100%, for the SF 10g 64% and 93%, for the copro-antigen ELISA 94% and 93%, for the ES ELISA 87% and 90% and for the Pourquier ELISA 88% and 84%. Significant between-season differences (P<0.05) were observed in the sensitivities of the two serological ELISAs: whereas the Pourquier ELISA had a higher sensitivity in spring than in autumn, the opposite was true for the ES ELISA. There were no significant between-season differences in the specificity for any of the tests. The test results of the SF 4g, copro-antigen ELISA and ES ELISA were associated with the level of infection of the animal. Given a positive test result of the SF 4g it is at least 11 times more likely that the animal is carrying a heavy infection (>10 flukes) than that is free of infection or lightly infected (< or = 10 flukes). Weak ( approximately 0.3) and moderate ( approximately 0.6) correlations were observed within infected animals of level of infection with ES and copro-antigen ELISA results, respectively. 相似文献
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蓝马鸡和藏马鸡的血清酯酶同工酶酶谱 总被引：2，自引：1，他引：1
采用聚丙烯酰胺凝胶垂直平板电泳法对 1 9只蓝马鸡和 1 5只藏马鸡的血清酯酶同工酶酶谱进行了研究。结果发现 :(1 ) 2种马鸡的血清酯酶存在ES1 ,ES2和ES3 3种同工酶 ;(2 )蓝马鸡的ES1同工酶存在显现酶活性的ES1A (36 8% )和不显现酶活性的ES1O (63 2 % ) 2种表型 ,而藏马鸡全部为ES1O型 ;(3)ES2同工酶只有一条浓染的酯酶区带 ;(4)ES3同工酶存在ES3ABC ,ES3AB和ES3BC 3种表型 ,蓝马鸡和藏马鸡均以ES3ABC为优势表型 (分别为 68 4%和 60 0 % )。 相似文献
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不同越冬饲料对蜜蜂中肠消化酶活性、组织发育状态以及抗氧化酶基因表达的影响 总被引：1，自引：0，他引：1
本试验旨在研究不同越冬饲料对蜜蜂中肠消化酶活性、组织发育状态以及抗氧化酶基因表达的影响,以便为广大蜂农选择越冬饲料提供参考。在2015年10月底选取群势相当的本地意大利蜜蜂(Apis mellifera L.)越冬蜂群(群内无储备越冬饲料脾)9群(5框蜂/群),随机分为3个试验组(3群/组),从11月2号开始分别以蜂蜜(蜂蜜组)、果葡糖浆(果葡糖浆组)和白砂糖水(白砂糖∶水=2∶1,蔗糖组)为越冬饲料进行饲喂,饲喂至11月下旬蜂群进入越冬期。整个试验期为2015年10月底至2016年3月初。分别于越冬前(11月初)、越冬中期(1月初)和越冬后(3月初)采集蜜蜂中肠,测定中肠消化酶(淀粉酶、蔗糖酶和蛋白酶)活性,在越冬中期采集蜜蜂中肠用于中肠组织发育状态和抗氧化酶基因[超氧化物歧化酶1(Sod1)、超氧化物歧化酶2(Sod2)和过氧化氢酶(CAT)]相对表达量等指标的检测。结果表明:在越冬中期,果葡糖浆组和蜂蜜组的蜜蜂中肠内淀粉酶活性显著高于蔗糖组(P0.05)。在越冬中期和越冬后,饲喂不同越冬饲料的蜜蜂中肠内蔗糖酶活性差异不显著(P0.05)。在越冬中期,果葡糖浆组的蜜蜂中肠内蛋白酶活性显著高于蔗糖组和蜂蜜组(P0.05)。蜂蜜组和蔗糖组的蜜蜂中肠肠壁厚度极显著大于果葡糖浆组(P0.01),且蜂蜜组和蔗糖组的蜜蜂中肠隐窝深度也极显著大于果葡糖浆组(P0.01)。在越冬中期,蔗糖组的蜜蜂中肠Sod1基因的相对表达量显著高于蜂蜜组和果葡糖浆组(P0.05)。由此得出,蜂蜜能够提高越冬蜂中肠消化酶活性,蜂蜜和白砂糖有利于越冬蜂中肠组织的发育,而且白砂糖能够提高越冬蜂中肠抗氧化酶基因的表达。因此,蜂蜜和白砂糖比果葡糖浆更适合作为蜜蜂的越冬饲料。 相似文献
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《Journal of Equine Veterinary Science》1996,16(8):334-338
Weight maintenance and blood chemistry parameters of geriatric (>20 yrs) horses and ponies housed at a farm for retired horses were compared between animals fed a 14% protein pelleted/extruded feed (ES) formulated specifically for geriatric horses versus those fed the textured sweet feed used at the facility. A total of 48 geriatric horses and 3 ponies were initially examined, 28 of which were assigned to be fed ES, the other 23 continued to be fed an 8.5% protein textured sweet feed mix (SF). Horses and ponies were fed the feeds in amounts estimated to be necessary for weight maintenance or gain according to initial body condition scores, with hay being offered free choice. The horses were weighed, assigned body condition scores and had blood drawn for complete blood counts and blood chemistry panels in December, 1992 before feed changes were initiated. The same measurements were taken on surviving horses and ponies three months after the feed changes had been made (March, 1993).For statistical analyses, data were grouped according to the animal's initial body condition score. Horses (n=17) and a pony with initial body condition scores of three or less (Group 1) which were fed ES had greater weight gain (ES: 32kg±4; SF: −9kg±7; p<.001); higher final condition scores (ES: 4.5±2; SF: 3.6±.3; p<.05) and plasma total protein (ES: 6.3 gm/dl±.2; SF: 5.2 gm/dl±.3; p<.005) in March than those fed SF (n=14). Group 1 horses fed SF experienced a decrease (p<.0001) in hematocrit (December: 41.4%±1.3; March: 34.4±1.2) and hemoglobin (December: 14.6±.4 gm/dl; March: 11.9±.3 gm/dl, p<.001), whereas those fed ES had a slight increase (p<.15) in hematocrit and no change (p>.2) in hemoglobin over the same period. Blood phosphorus was increased (December: 2.21±.08 mg/dl; March: 2.48±.07 mg/dl; p<.05) in Group 1 horses fed ES and tended (p<.15) to be higher than in Group 1 horses fed SF (March: 2.27±.12 mg/dl). In horses and the pony with initial condition scores greater than 3 (Group 2), condition scores, body weights and hematocrit did not differ (p>.2) between animals fed ES (n-9) versus SF (n=9) in December or March. However Group 1 animals fed ES experienced greater weight gains (p<.001) and increases in body condition score (p<.05) than Group 2 SF fed horses during this period. Blood phosphorus in Group 2 ES fed horses tended to increase (December: 2.3±.1 mg/dl; March: 2.6±.1 mg/dl; p<.10) and was higher (p<.01) in March than in Group 2 horses fed SF (March: 2.1±.1 mg/dl). In both groups, liver enzymes and creatinine did not differ (p>.2) between diets in March. Blood urea nitrogen (BUN) was higher in ES fed horses than the SF fed horses in March in both Group 1 (p<.02) and Group 2 (p<.07) but was within the normal range for adult horses.Horses which were fed the ES were subjectively assessed by the farm manager to have better hair coats and higher activity levels than when they had been fed textured sweet feed and relative to horses that were not fed ES. Geriatric horses, especially those unable to maintain adequate weight on standard rations in the absence of hepatic or renal disease, appear to benefit from the special formulation of ES. 相似文献
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白唇鹿血清同工酶酶谱的电泳研究 总被引：1，自引：1，他引：0
采用聚丙烯酰胺凝胶电泳地对３４头白唇夜以继日的４种血清同工酶酶谱及其多态性进行研究。结果（１）酯酶包括ＥＳ１，ＥＳ２和ＥＳ３三种同工酶，其中ＥＳ２存在多态性；（２）碱性磷酸酶存在在ＡＬＰＢ和ＡＬＰＯ两种表型，以ＡＬＰＯ型为优势表型（６７．６５％）；（３）淀粉酶包括ＡＭＹ１，ＡＭＹ２和ＡＭＹ３三种同工酶，ＡＭＹ１和ＭＡＹ２两种同工酶存在多态性，（４）白唇鹿的乳酸脱氢酶显现出与众不同的同工酶酶谱。 相似文献