三氮脒含量测定的高效液相色谱法研究

建立了HPLC法测定注射用三氮脒中三氮脒含量的方法。采用C18色谱柱,以0.05 mol/L磷酸溶液-乙腈(95∶5,V/V)为流动相,流速为1.0 m L/min,检测波长为370 nm,柱温为30℃。结果三氮脒在1~100μg/m L浓度范围内线性关系良好,相关系数r=0.9998。建立的方法操作简便、准确、可靠,可用于三氮脒的质量控制。     

三氮脒与含苯磺酰胺结构单元的新型三氮脒衍生物对小鼠急性毒性试验

为了比较三氮脒及含苯磺酰胺结构单元的新型三氮脒衍生物在不同给药方式下对小鼠的毒性,以评估两种药物的安全性,根据改良寇氏法测定三氮脒及三氮脒衍生物在不同给药方式下对小鼠的半数致死量(LD_(50))及95%的可信限,给药方式分为腹腔注射和皮下注射两种。试验结果显示,腹腔注射时三氮脒和新型衍生物的LD_(50)分别为235.0 mg/kg、251.8 mg/kg,95%可信限为157.0～351.7 mg/kg、168.6～376.0 mg/kg;皮下注射时三氮脒的LD_(50)为257.6 mg/kg,95%可信限为171.2～387.8 mg/kg,新型衍生物的LD_(50)500.0 mg/kg。以上试验结果表明,新合成的衍生物其毒性和三氮脒相比,仅皮下注射时LD_(50)显著降低。对含苯磺酰胺结构单元的新型三氮脒衍生物的研发将具有重要的意义。     

三氮脒注射液在山羊体内的药动学比较

研究新制剂三氮脒注射液在山羊体内的药动学特征。用三氮脒和三氮脒注射液给山羊单剂量(3.5mg·kg-1)肌内注射,采用高效液相色谱法测定山羊体内三氮脒含量,3P97软件拟合血药浓度-时间曲线,计算药动学参数。三氮脒和三氮脒注射液药动学参数符合一级吸收二室模型,主要药动学参数分别为:达峰浓度(Cmax)为(5.97±0.37)μg·mL-1和(7.63±0.41)μg·mL-1;达峰时间(Tmax)为(0.22±0.03)h和(0.40±0.10)h;吸收半衰期(t1/2ka)为(0.13±0.04)h和(0.04±0.01)h;药时曲线下面积(AUC)为(53.96±6.87)μg·mL-1*h和(77.43±6.71)μg·mL-1*h。三氮脒注射液与三氮脒相比,具有吸收缓慢、分布广、安全性好和作用时间长等特点,此结论为临床合理使用剂型提供依据和指导。     

山羊血浆中三苯双脒的高效液相色谱检测方法的研究

建立了山羊血浆中三苯双脒的高效液相色谱检测方法.山羊血浆用氢氧化钠和三乙铵处理,再用甲苯萃取三苯双脒,高速离心后,上清液真空干燥,残留物用含内标物的甲醇溶解后分析.采用Waters C18色谱分析柱(3.9 mm×150 mm,10 μm),流动相为甲醇-水(50:50,pH 3),流速0.6 mL/min,UV检测波长249 nm,柱温为25℃,内标物为水杨酸(5μg/mL).测得三苯双脒保留时间为4.3 min,水杨酸保留时间为8.0 min.在50～1 000 ng/mL浓度范围内三苯双脒和内标物峰面积之比与三苯双脒浓度具有良好的线性关系,回归方程为A=1 456.67C-60.01(r=0.974 7,n=5),RSD为5.1%.1 000、500和100 ng/mL质控浓度添加样品的日内(n=5)和日间(n=5)RSD分别为3.8%、9.7%、11.7%和5.2%、8.0%、11.1%.回收率为(94.3±1.1)%、(91.9±0.9)%和(94.7±5.0)%.最低检测限为20 ng/mL.本方法操作简便,灵敏度高,为三苯双脒兽用制剂临床前及临床药物代谢动力学研究提供了分析方法.     

三氮脒和强力霉素对猪附红细胞体病疗效的比较试验

为研究三氮脒和强力霉素对猪附红细胞体病的治疗效果,为该病防治提供参考依据。本试验共分为2个试验组和1个对照组,仔猪舍之间完全隔开,30头/组,对试验组和对照组红细胞进行镜检,并进行胆红素代谢试验以及三氮脒和强力霉素治疗对比试验。结果发现,发病猪红细胞周围呈刺状或无规则形状,可不规则移动,在血浆中可见到游离活附红细胞体。治疗后,血浆中游离的附红细胞体消失。发病猪血清TBIL和IBIL明显升高,治疗后,与对照组相比,治疗组TBIL和IBIL明显下降(P0.05),三氮脒组和强力霉素组之间TBIL存在显著差异(P0.05),并且强力霉素组IBIL降低至正常范围内(3.51μmol/L)。得出结论,附红细胞体病可引起发病猪只IBIL显著升高,三氮脒或强力霉素均有较好的治疗效果,且强力霉素较三氮脒效果好。     

氟腺嘌呤原料药中有关物质检查及含量测定的HPLC法

为了建立氟腺嘌呤原料药中有关物质检查及含量测定的高效液相色谱法,采用色谱柱Unitary C18(4.6 mm×250 mm,5.0μm),流动相为乙腈-水=40∶60(v/v),流速为1.0 mL/min,柱温为30℃,紫外检测波长为261 nm。结果:氟腺嘌呤在50¹00μg/mL的浓度范围内,峰面积与浓度呈良好线性关系(r=0.999 8,n=5)。精密度试验RSD=0.66%,稳定性试验(90 d)RSD=1.86%。定量限为24 ng/mL,检测限为64 ng/mL。有关物质检查中杂质总量均未超过1.0%。该方法为氟腺嘌呤的含量测定和有关物质检查提供了方便、灵敏、精确、可靠的方法,为氟腺嘌呤的质量控制提供依据。     

“复方三氮脒注射液”治疗猪附红细胞体病临床疗效观察

以三氮脒、磺胺二甲氧嘧啶钠注射液为对照,用高、中、低剂量的"复方三氮脒注射液"对临床诊断为附红细胞体病的病猪进行临床疗效观察。结果高、中、低剂量的"复方三氮脒注射液"对猪附红细胞体病的有效率分别为100%、100%、96.67%,治愈率分别为98.33%、95.00%和85.00%;有效率和治愈率均明显高于三氮脒、磺胺类药物注射液对照组,差异极显著(P<0.01)。临床推荐使用剂量为中剂量。     

北京地区25例犬吉氏巴贝斯虫病例分析

为调查北京地区犬吉氏巴贝斯虫病的感染与治疗情况,2014年10月至2016年1月期间,本试验对中国农业大学动物医院就诊的25例犬吉氏巴贝斯虫感染病例进行调查统计,记录其临床症状,分析全血细胞计数(Complete Blood Count,CBC)、生化检测结果和三氮脒的治疗效果。结果显示,25例患犬均出现不同程度的贫血,73.3%(11/15)的患犬出现了免疫介导性溶血性贫血(Immune mediated hemolytic anemia,IMHA),使用三氮脒和输血疗法治疗后,71.2%(10/14)的患犬取得良好的疗效,但目前跟踪的病例中,21.4%(3/14)的患犬已复发。     

中西医结合治疗羊泰勒虫病效果观察试验研究

本试验对"碘硝酚+三氮脒+当归补血汤"中西医结合的治疗方法与羊泰勒虫病较敏感的三氮脒、黄色素进行了疗效对比。结果表明应用三氮脒、黄色素和中西医结合治疗羊泰勒虫病治愈率分别为78.46%、56.43%和92.4%。"碘硝酚+三氮脒+当归补血汤"组的治愈率均极显著地高于其他两组(P0.01),这种方法不仅能提高发病后期羊的治愈率,而且可缩短治疗周期,加快羊的体质恢复,具有很高的临床价值。     

紫外分光光度法测定三氮脒含量

三氮脒为抗原虫药 ,主要用于防治家畜巴贝斯焦虫病、泰勒氏焦虫病。该药为《中国兽药典》所收载 ,由兰州中亚药械厂独家生产。其含量测定采用传统的永停滴定法 ,测定条件要求严格 ,不易控制和掌握。笔者利用三氮脒在紫外区有明显的吸收峰的特点 ,成功地对三氮脒含量进行测定 ,取得了满意的结果。1　实验材料1.1　仪器岛津 UV- 2 5 0 1紫外分光光度计。1.2　试剂蒸馏水。1.3　对照品三氮脒原料精制品 ,含量 99.8%。1.4　样品三氮脒成品 ,由兰州中亚药械厂库房随机抽取。2　方法与结果2 .1　测定波长的选择精确称取三氮脒细粉适量 ,加蒸馏水…     

Effect of tetracycline administration on the efficacy of diminazene aceturate therapy and prophylaxis in Trypanosoma brucei infections of mice

The simultaneous administration, to Trypanosoma brucei infected mice, of rolitetracycline or oxytetracycline and diminazene aceturate appeared to have no effect on the initial trypanocidal action of the diminazene aceturate in that trypanosomes were cleared from the circulation. It also had no effect on the duration of the aparasitaemic period which follows diminazene aceturate treatment and the mice remained free of circulating trypanosomes for some time. However, if used prophylactically, relatively large amounts of tetracyclines (4 x 10 mg kg-1) administered with 40 mg kg-1 diminazene aceturate caused a reduction of the prophylactic period compared with those mice given only diminazene aceturate. This reduction in the prophylactic period is unlikely to have any practical significance in the combination diminazene aceturate/tetracycline treatment of domestic animals as diminazene aceturate is used therapeutically and not prophylactically in the control of trypanosomiasis.     

附红清治疗小白鼠人工感染猪附红细胞体试验观察

将人工感染附红细胞体小白鼠50只分为附红清治疗高、中、低剂量组和三氮脒治疗对照组及生理盐水对照组,采用腹腔注射给药,1次/d,连用3 d,连续6 d采血观察红细胞感染率。结果附红清高、中剂量组与三氮脒对照组疗效接近。     

The Effect of Diminazene Aceturate on Spienectomized Dogs with Babesia gibsoni Infection

The effect of diminazene aceturate on splenectomized and nonspienectomized dogs with Babesia gibsoni (B. gibsoni) infection was investigated. In splenectomized dogs, the fissional and multiplicational stages of B. gibsoni were observed in peripheral blood films, and hemoglobinuria was frequently observed. These findings were different from previous reports and were not changed by administration of diminazene aceturate. It is clear that the intramuscular administration of diminazene aceturate at the dose of 3 mg/kg body weight for 3 days is not effective against B. gibsoni infection in splenectomized dogs.     

Treatment of experimental trypanosomiasis in pigs.

The therapeutic activity of difluoromethylornithine (DFMO), diminazene aceturate (Berenil) and their combination against chronic trypanosomiasis was investigated in experimental Trypanosoma brucei brucei infections of growing pigs. DFMO (300 mg/kg/day orally for 10 days), diminazene aceturate (7 mg/kg in single intramuscular injection) and a combination of the two agents at the above dosages produced varied periods of aparasitaemia in the treated pigs. Relapse parasitaemia occurred in all treatment groups, with diminazene aceturate providing the longest relief period of 17 days, combination treatment 11 days and DFMO 6 days. The packed cell volume, blood haemoglobin concentration and red cell count values decreased after the pigs were infected with the parasites. The values improved following treatment with the agents and their combination.     

Comparative efficacy of diminazene diaceturate and diminazene aceturate for the treatment of babesiosis in horses

The present work was conducted to study the comparative efficacy of two anti-protozoan (babesicidal) drugs on the recovery and health of horses. A total of 80 horses, showing typical clinical symptoms of the disease, were selected for this study; the presence of babesiosis was confirmed through blood smear examination. These animals were divided into two groups i.e. A and B. Horses of Group A, were treated with diminazene diaceturate, while horses of Group B were treated with diminazene aceturate. Efficacy of the drugs was determined by the reversal of clinical signs and a negative blood smear examination. The efficacy of diminazene diaceturate was demonstrated to be 80% while diminazene aceturate was found to have 90% efficacy against babesiosis.     

HPLC-ECD法测定大观霉素的含量

建立了HPLC—ECD法测定大观霉素含量的分析方法。采用高效液相色谱-电化学检测器,Waters XBridge C18色谱柱（4．6mm×250mm,5μm）,以pH3．2的革酸盐缓冲溶液-乙腈（1000：105,V：V）为流动相,流速1．0mL／min;柱后衍生溶液为21g／L的NaOH溶液,流速0．5mL／min;ECD系统参数：极性为阳性,检波＋0．12V,氧化电位＋0．70V,还原电位-0．60V,范围5μA。结果表明,大观霉素及4R一二氢大观霉素在40～320μg／mL范围内呈良好的线性关系（大观霉素r=0．9999,4R-二氢大观霉素r=0．9992）,平均回收率为100．0％（n=9）,RSD为0．2％。该方法解决了微生物效价测定大观霉素含量偏高的难题,且操作简便、结果准确,可用于大观霉素及相关制剂产品的质量控制。     

The pharmacokinetics of diminazene aceturate after intramuscular administration in healthy dogs

The pharmacokinetics of diminazene aceturate following intramuscular (i.m.) administration at 4.2 mg/kg was evaluated in 8 healthy German Shepherd dogs. Blood samples were collected at 19 intervals over a period of 21 days. Diminazene plasma concentrations were measured using a validated HPLC method with UV detection and a sensitivity of 25 ng/ml. The in vitro and in vivo binding of diminazene to blood elements was additionally determined. Diminazene pharmacokinetics showed a large inter-individual variation after i.m. administration. It had a short absorption half-life (K01-HL of 0.11 +/- 0.18 h), resulting in a C(max) of 1849 +/- 268.7 ng/ml at T(max) of 0.37 h and a mean overall elimination half-life (T1/2beta) of 5.31 +/- 3.89 h. A terminal half-life of 27.5 +/- 25.0 h was measured. At 1 h after i.m. injection, 75% of the diminazene in whole blood was in the plasma fraction. The results of this study indicate that diminazene is rapidly distributed and sequestered into the liver, followed by a slower terminal phase during which diminazene is both redistributed to the peripheral tissues and/or renally excreted. It is recommended that diminazene administered i.m. at 4.2 mg/kg should not be repeated within a 21-day period.     

Response of Trypanosoma congolense in goats to single and double treatment with diminazene aceturate.

Diminazene aceturate is one of a limited number of compounds currently marketed for treatment of trypanosomiasis in cattle, sheep and goats. The pharmacokinetics of the compound in goats suggest that double treatment with diminazene aceturate might enhance the compound's therapeutic activity. A study was therefore conducted in goats using two clones of Trypanosoma congolense, IL 3274 and IL 1180, which were previously shown to be resistant and sensitive, respectively, to single treatment with diminazene aceturate. The results indicated that, as compared to single treatment, double treatment with diminazene aceturate at a dose of 7.2 mg kg-1 bodyweight, at either eight or 24 hour intervals, did not greatly enhance the therapeutic activity of the drug. Furthermore, treatment with the same drug dose eliminated infections with T congolense IL 3274 when treatment was administered 24 hours after infected Glossina morsitans centralis had fed, but failed to do so if treatment was delayed until after goats were detected to be parasitaemic. This suggests that failure of T congolense IL 3274 to respond to treatment with diminazene may not be due to drug resistance per se.     

吡喹酮注射剂含量测定方法比较

采用两种不同的色谱条件测定吡喹酮注射剂中吡喹酮的含量,色谱柱为迪马DiamonsilC18（4．6mm×250mm,5μm）柱,流动相为甲醇-水及乙腈-水,检测波长为263nm及210nm,流速1mL/min。结果显示,在两个不同的检测范围内,线性关系均良好（分别为r=0．9998和r=0．9996）,吡喹酮的平均回收率为100．38％和99．90％,RSD=1．13％和1．1％（n=9）。两种方法简便易行,精密度、回收率、重现性均符合要求。结果表明,两种方法均可用于吡喹酮注射剂的含量测定。     

Concentrations of isometamidium chloride (Samorin®) in sera of Zebu cattle which showed evidence of hepatotoxicity following frequent trypanocidal treatments

The concentrations of isometamidium circulating in poorly nourished Zebu cattle which showed morbidity, mortality, and biochemical and histopathological evidence of hepatotoxicity, following frequent treatments with isometamidium chloride and diminazene aceturate were investigated using the isometamidium-ELISA. As few as two isometamidium treatments one month apart were associated with significant weight loss, and cattle treated with diminazene aceturate after three or four isometamidium treatments suffered a 50% mortality.
Although there were no obvious, marked elevations in isometamidium concentration which might have allowed the use of the ELISA as a predictor of a potential toxicity problem, concentrations did increase significantly with the number of monthly treatments administered, suggesting drug accumulation, and the increases were significantly higher in cattle to which diminazene had also been administered. In cattle treated with both trypanocides, weight loss and serum glutamate dehydrogenase levels were correlated with isometamidium concentrations. These observations, together with the histopathological findings, support the hypothesis that the morbidity and mortality observed were related to the repeated treatment with isometamidium in conjunction with diminazene aceturate, and that the pathogenesis involved a component of hepatic damage.
It is therefore recommended that cattle, particularly those under nutritional stress, are not subjected to repeated treatments with isometamidium at intervals as short as one month, and particularly not with concurrent administration of diminazene.