UFLC-MS/MS法同时测定枳实中6个活性成分的含量

为了建立超快速液相色谱-串联质谱法同时测定枳实中6个活性成分(异柚皮苷、柚皮苷、橙皮苷、新橙皮苷、柚皮素和橙皮素)的含量,采用Shim-Pack XR-ODS柱(75 mm×3.0 mm,2.2μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱(0～10 min,10%→30%B;10～12 min,30%→60%B;12～15 min,60%→85%B),平衡时间为3 min,流速为0.5 mL/min,柱温为30℃,进样量为5μL。质谱采用3200 QTRAP三重四极杆串联质谱仪,离子源为电喷雾电离源(ESI源),检测方式为负离子检测,扫描模式为多重反应检测(MRM)。结果显示,异柚皮苷、柚皮苷、橙皮苷、新橙皮苷、柚皮素和橙皮素浓度分别在0.2～8.0μg/mL(r=0.999 2)、1.0～40μg/mL (r=0.999 4)、0.2～8.0μg/mL (r=0.999 7)、0.5～20μg/mL(r=0.999 2)、0.02～0.8μg/mL (r=0.999 7)和0.02～0.8μg/mL (r=0.999 7)浓度范围内与峰面积呈现良好的线性关系;平均回收率(n=9)分别为99.2%、98.7%、98.6%、98.8%、99.3%和99.1%。6批枳实样品测定结果显示:异柚皮苷1.70%～1.97%,柚皮苷11.6%～12.4%,橙皮苷0.481%～0.576%,新橙皮苷3.15%～4.13%,柚皮素0.293%～0.391%和橙皮素0.363%～0.395%。表明该方法快速、准确、重复性好,可用于枳实的质量控制,同时为新兽药开发利用提供参考。     

QuEChERS/高效液相色谱串联质谱法测定渔业饲料中敌百虫、敌敌畏、蝇毒磷的含量

本文建立了一种简单、快速测定渔业饲料中敌百虫、敌敌畏、蝇毒磷的QuEChERS-液相色谱串联质谱分析方法。样品经酸化乙腈提取,C18与中性氧化铝净化后,采用HPLC-MS/MS选择反应监测(SRM)正离子模式测定进行定性、定量分析。结果表明:在2.0～200 ng/mL范围内,峰面积与浓度线性良好,相关系数均大于0.9992;方法的检出限(LOD)为2.0μg/kg,定量限(LOQ)为4.0μg/kg,检测结果的相对标准偏差为1.62%～8.68%(n=6),平均加标回收率82.3%～105.2%。该方法具有较高的重现性和选择性,适用于渔业饲料中敌百虫、敌敌畏、蝇毒磷的残留分析检测。     

牛奶中有机磷农药残留的气相色谱法测定

提出同时测定牛奶中敌百虫、甲胺磷、敌敌畏、马拉硫磷、倍硫磷五种有机磷农药残留的检测方法。在选定的色谱条件下,添加浓度在0.5～10.0μg/mL之间时5种农药的检测限分别为0.031μg/mL、0.013μg/mL、0.023μg/mL、0.019μg/mL和0.014μg/mL,回收率范围分别达81.0%～85.5%、84.5%～89.4%、79.0%～81.8%、77.3%～82.5%和77.9%～89.5%,变异系数在0.29%～3.96%之间。本方法对牛奶中有机磷农药残留的检测分析准确可靠。     

梯度洗脱HPLC法同时测定4种解热镇痛类药物的含量

建立梯度洗脱HPLC法同时测定4种解热镇痛类药物的含量方法。采用十八烷基硅烷键合硅胶色谱柱(4.6 mm×250 mm,5μm),以甲醇-0.05 mol/L醋酸铵溶液为流动相进行梯度洗脱,流速1.0 mL/min,柱温为30℃,检测波长254 nm。结果表明,在该色谱条件下,对乙酰氨基酚、安乃近、安替比林、氨基比林分离良好;对乙酰氨基酚在50～500μg/mL范围内线性良好,R=0.999 8,平均回收率为99.2%(n=9),RSD为0.6%;安乃近在10～250μg/mL范围内线性良好,R=0.999 8,平均回收率为100.6%(n=9),RSD为0.8%;安替比林在10～100μg/mL范围内线性良好,R=0.999 6,平均回收率为99.4%(n=9),RSD为1.0%;氨基比林在10～250μg/mL范围内线性良好,R=0.999 7,平均回收率为98.9%(n=9),RSD为0.8%。本测定方法操作简便,结果准确、可靠,可用于同时测定以上4种解热镇痛类药物的含量。     

毛细管胶束电动色谱法检测饲料中防腐剂

建立了同时分离测定山梨酸(SA)、苯甲酸(BA)、对羟基苯甲酸甲酯(MP)、对羟基苯甲酸乙酯(EP)、对羟基苯甲酸丙酯(PP)、对羟基苯甲酸丁酯(BP)6种防腐剂的胶束电动毛细管色谱方法。以十二烷基硫酸钠(SDS)-磷酸盐-硼砂缓冲体系为缓冲液,pH为7.73,电压为20 kV,于245 nm波长下对样品进行检测。结果表明,6种防腐剂均在14 min内实现很好分离,SA、BA的线性范围分别为0.5～150μg/mL和15～200μg/mL,MP、EP的线性范围均为2～200μg/mL,PP的线性范围为5～200μg/mL,BP的线性范围为1～200μg/mL,线性相关系数为0.9987～0.9995,平均回收率为89.7%～96.7%,RSD≤5.91%,检测限为0.25～10μg/mL。     

HPLC法测定鸡肉中恩诺沙星、环丙沙星残留的研究及应用

建立高效液相色谱(HPLC法)测定动物组织中恩诺沙星、环丙沙星的残留,具有很好的分离度,浓度5～500μg/mL呈良好的线性关系,R=0.99998;仪器检出限为5～10μg/mL;回收率范围为80.6%～99.7%;批内变异系数≤2.6%,批间变异系数≤7.6%。检测50批鸡肉中恩诺沙星、环丙沙星的残留,检出率为16.0%,检出的样品中,残留的量范围为9.04～79.7μg/kg,无超标现象。     

绒山羊瘤胃内环境参数及血清中微量元素的测定

对36只健康放牧绒山羊的瘤胃内环境参数及血清中微量元素含量进行了测定。瘤胃内环境参数:pH值6.57±0.33,总酸度(33.66±6.18)U,氨氮浓度(3.51±1.10)mg/100mL,活纤毛虫数(38.93±7.54)万/mL,总纤毛虫数(53.00±19.97)万/mL,纤毛虫活率(66.54±12.07)%,细菌以革兰氏阴性为主。瘤胃液中微量元素含量为:磷(5.35±1.39)μg/mL,钾(889.40±224.67)μg/mL,钠(2423.79±495.22)μg/mL,钙(78.34±28.32)μg/mL,镁(34.31±8.44)μg/mL,铜(1.43±0.55)μg/mL,锌(4.81±1.73)μg/mL,钴(0.55±0.16)μg/mL,硒(0.10±0.05)μg/mL。血清中微量元素含量为:钾(139.91±67.51)μg/mL,钠(3273.75±579.79)μg/mL,钙(45.01±17.28)μg/mL,镁(23.28±8.22)μg/mL,铜(1.42±)μg/mL,锌(5.02±)μg/mL,1.001.84钴(0.51±0.19)μg/mL,硒(0.30±)0.13μg/mL。     

紫外分光光度法测定盐酸沙拉沙星可溶性粉中沙拉沙星方法的建立

为了快捷简便地测定盐酸沙拉沙星可溶性粉生产过程中沙拉沙星的含量,采用紫外分光光度法测定沙拉沙星的含量。结果表明,沙拉沙星对照品质量浓度在2.4μg/mL～12.0μg/mL范围内与吸收度呈良好的线性关系,线性回归方程A=0.088C+0.09,r=0.9999(n=5),平均回收率为99.74%,RSD=0.47%(n=5),与高效液相色谱法比较结果一致。     

HPLC法同时测定清胃润肠合剂中4种成分含量

建立高效液相色谱法同时测定清胃润肠合剂中芒果苷、盐酸巴马汀、盐酸小檗碱、甘草酸含量的方法。采用Welchrom?C₁₈(4.6 mm×250 mm,5μm)色谱柱,流动相为乙腈-0.05 mol/L磷酸二氢钾溶液(用磷酸调节pH值至3.5),梯度洗脱,流速1.0 mL/min,柱温30℃,进样量10μL,检测波长258 nm。结果表明,芒果苷、盐酸巴马汀、盐酸小檗碱、甘草酸分别在13.600～217.550μg/mL、9.410～150.560μg/mL、35.670～570.690μg/mL、40.910～654.560μg/mL范围内线性关系均良好(r>0.999 0),平均加样回收率(n=6)分别为102.88%、102.60%、99.50%、100.77%,RSD分别为0.49%、0.91%、0.49%、0.41%。该方法快速简便、精密度好、灵敏度高,可用于清胃润肠合剂的质量控制。     

高效毛细管电泳法同时检测饲料中六种抗氧化剂

实验以25 mmol/L硼砂-硼酸(Na2B4O7-H3BO3)和10 mmol/L十二烷基硫酸钠(SDS)的混合液(pH=7.00)作为缓冲液,采用高效毛细管胶束电动色谱法测定样品中丁基羟基茴香醚(BHA)、二叔丁基甲基苯酚(BHT)、特丁基对苯二酚(TBHQ)、没食子酸异丙酯(PG)、没食子酸辛酯(OG)、乙氧基喹啉(EQ)六种抗氧化剂。该方法在15 min内实现了六种抗氧化剂的分离;TBHQ和BHA的线性范围为50～500μg/mL,PG、EQ、OG和BHT的线性范围分别为10～200、20～200、25～400μg/mL和25～500μg/mL,线性相关系数≥0.996。TBHQ、PG、BHA、OG、EQ、BHT的检测限分别为30、5、25、15、10、10μg/mL;样品平均回收率为81.8%～102.0%,相对标准偏差≤6.5%。     

白术、微米白术和白术多糖对断奶仔猪生长性能和肠道形态及微生态区系的影响

试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。     

Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia

Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.     

Phase I evaluation of CCNU (lomustine) in tumor-bearing cats

1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.     

Prevalence of gastric lesions in racing Alaskan sled dogs

Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large.     

Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects

Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.     

Y‐chromosomal variation of local goat breeds of Turkey close to the domestication centre

Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.     

The Epidemiology of Gastrointestinal Nematodes of Dairy Cattle in Central Kenya

The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.     

Diversity of Ectoparasites in Sheep Flocks in Sa~o Paulo,Brazil

The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.     

Feline intracranial neoplasia: retrospective review of 160 cases (1985-2001)

The purpose of this study was to determine the frequency of different tumor types within a large cohort of cats with intracranial neoplasia and to attempt to correlate signalment, tumor size and location, and survival time for each tumor. Medical records of 160 cats with confirmed intracranial neoplasia evaluated between 1985 and 2001 were reviewed. Parameters evaluated included age, sex, breed, FeLV/FIV status, clinical signs, duration of signs, number of tumors, tumor location(s), imaging results, treatment, survival times, and histopathologic diagnosis. Most of the cats were older (11.3 +/- 3.8 years). Primary tumors accounted for 70.6% of cases. Metastasis and direct extension of secondary tumors accounted for only 5.6 and 3.8% of cases, respectively. Twelve cats (7.5%) had 2 or more discrete tumors of the same type, whereas 16 cats (10.0%) had 2 different types of intracranial tumors. The most common tumor types were meningioma (n = 93, 58.1%), lymphoma (n = 23, 14.4%), pituitary tumors (n = 14, 8.8%), and gliomas (n = 12, 7.5%). The most common neurological signs were altered consciousness (n = 42, 26.2%), circling (n = 36, 22.5%), and seizures (n = 36, 22.5%). Cats without specific neurological signs were common (n = 34, 21.2%). The tumor was considered an incidental finding in 30 (18.8%) cats. In addition to expected relationships (eg, meninges and meningioma, pituitary and pituitary tumors), we found that lesion location was predictive of tumor type with diffuse cerebral or brainstem involvement predictive of lymphoma and third ventricle involvement predictive of meningioma.