高效液相色谱法测定兽用复方酮康唑乳膏中酮康唑的含量

笔者建立了高效液相色谱法测定兽用复方酮康唑乳膏中酮康唑含量。样品经80℃水浴用乙醇提取等前处理,采用Agilent ZORBAX Eclipse(4.6 mm×150 mm,5μm)色谱柱;流动相为甲醇-水（74∶26）;流速为1.0 mL/min;柱温为30℃;检测波长为239 nm。结果酮康唑在0.4015～1.2016 mg/mL范围内呈良好线性,r2=1.000。样品平均回收率为99.52%,RSD为0.70%。该法简便可靠,可测定兽用复方酮康唑乳膏中酮康唑含量。     

复方酮康唑纳米乳的研制及其体外经皮吸收

本研究研制出一种以纳米乳为载体的质量稳定,高效的药物传递系统.通过伪三元相图法筛选配方,找到最佳配比;通过HPLC法及稳定性试验考察复方酮康唑纳米乳的稳定性;用大鼠皮肤及透皮扩散仪进行透皮试验,考察其体外透皮性能.复方酮康唑纳米乳是由EL-40,乙醇,2-苯乙醇,乙酸乙酯,酮康唑,丙酸氯倍他索,丁香酚,水构成的淡黄色澄清透明的液体,稳定性良好;其透皮速率与2.5％氮酮组相当,优于混悬液及复方酮康唑软膏.复方酮康唑纳米乳是一种质量稳定、高效的药物传递系统.     

高效液相色谱法测定复方酮康唑溶液中酮康唑的含量

在中国，目前仅北京市登记养犬数就高达50万以上，按一般发病比例至少也有5—10万的犬和相当数量的猫需要防治耳病，临床需求紧迫，市场前景广阔。但是，现有进口药剂尚不能完全解决各种复杂的耳病，也未见有适合中国国情的、具有中国特色的用于耳病防治的宠物专用药品正式生产销售。根据这种情况，本着弘扬民族兽医文化，服务临床和振兴民族兽药工业的宗旨，笔者充分发挥中西医结合的优势研制开发复方酮康唑溶液，弥补了我们在这一方面的不足，填补了此项空白。为了能够有效的控制该产品质量，制定了用HPLC法测定该产品中的酮康唑含量的方法。经实验证实，此方法灵敏度高，准确性好，操作简便易行，并且与其他组分得到了很好的分离，各组分间无干扰。     

维生素C对酮康唑抑制白色念珠菌的影响研究

为研究酮康唑治疗畜禽真菌感染过程中能否给与维生素C辅助治疗,体外测试了添加维生素C后白念珠菌对酮康唑的敏感性。结果发现,维生素C能显著抑制酮康唑对白色念珠菌的敏感性,为科学治疗畜禽机会性真菌临床感染提供了依据。     

HPLC-MS/MS法测定饲料中甲硝唑和二甲硝咪唑含量

建立了高效液相色谱-串联质谱法(HPLC-MS/MS)快速测定饲料中甲硝唑和二甲硝咪唑的含量。饲料样品用乙酸乙酯提取后,经PCX固相萃取柱净化处理,过0.22μm滤膜,以乙腈和含0.1%的甲酸水溶液为流动相,经Eclipse Plus C18色谱柱(3.0 mm×100 mm,1.8μm)分离,多级反应监测(MRM)正离子模式下扫描分析。结果表明,甲硝唑和二甲硝咪唑在0.2~50μg/L范围呈良好线性,线性相关系数均大于0.997,检出限均为0.2μg/kg,定量下限均为0.5μg/kg。样品在定量下限1倍、2倍、10倍3个加标水平下的平均回收率为95.3%~103.2%,相对标准偏差(RSD)为4.67%~8.46%。该方法灵敏、简便、准确,可用于饲料中甲硝唑和二甲硝咪唑的检测分析。     

酮康唑水分散软颗粒剂的制备及体外溶出度考察

酮康唑(Ketoconazole,KET)是20世纪70年代末由比利时杨森制药公司人工合成的第3代咪唑类广谱抗真菌药物。它通过干扰真菌细胞膜麦角甾醇的生物合成,影响细胞膜通透性而抑制其生长[1,2]。临床用于治疗各种不同类型的皮肤真菌病、阴道念珠菌病以及用于治疗和预防全身真菌感染和胃     

固相萃取结合液相色谱-串联质谱技术同时测定饲料中氯苯那敏和溴苯那敏含量

本试验旨在建立一种固相萃取结合液相色谱-串联质谱技术同时测定饲料中氯苯那敏和溴苯那敏含量的方法。饲料样品用1%甲酸乙腈溶液提取,经混合型阳离子固相萃取柱(MCX)净化,采用液相色谱-串联质谱测定,以0.1%甲酸溶液(A)-乙腈(B)为流动相,梯度洗脱,C18色谱柱分离。通过使用不同饲料制备基质添加标准曲线,氯苯那敏和溴苯那敏含量在1.0～100.0 ng/mL内线性关系良好(R²> 0.995 0);方法检出限为0.3μg/kg,定量限为1.0μg/kg;氯苯那敏和溴苯那敏在3个添加水平(1.0、10.0、100.0μg/kg)下的平均回收率为85.8%～108.4%,相对标准偏差(RSD,n=6)<15%。该方法前处理简单,操作方便,适用于饲料中氯苯那敏和溴苯那敏含量的同步测定。     

离子对高效液相色谱法测定复方甲硝唑片中甲硝唑和维生素B6含量

建立了同时测定复方甲硝唑片中甲硝唑和维生素B6含量的离子对HPLC方法.采用ODS色谱柱,流动相为磷酸二氢钾和庚烷磺酸钠水溶液(每1L中含磷酸二氢钾13.6 g与庚烷磺酸钠1.0 g)-甲醇-三乙胺(85∶15∶0.4),用磷酸调pH至2.8;检测波长290 nm,以峰面积计算,外标法定量.甲硝唑和维生素B6的线性范围分别为100～300 μg/mL(r=0.999 9)和5～50 μg/mL(r=0.999 9);平均回收率分别为100.2%(RSD=1.0%,n=6),99.8%(RSD=1.2%,n=6).该方法测定结果准确,操作简便,回收率高.     

加米霉素注射液含量的高效液相色谱检测法的建立

为了建立加米霉素注射液含量的高效液相色谱检测法,为其检测提供依据,试验选用样品浓度为0.3 mg/mL,进样量为20μL,流速为1.0 mL/min,柱温为40℃,溶剂为乙腈,作为初步的色谱条件,比较不同比例流动相、不同色谱柱的峰形及分离效果,确定最优色谱条件,并考察该方法的线性关系、重复性、稳定性及添加回收率,并利用该方法对样品进行检测。结果表明:通过筛选确定了液相色谱检测条件,色谱柱为资生堂CAPCELL PAK C_(18) MGⅡ(250 mm×4.6 mm,5μm),流动相为磷酸盐缓冲液(取0.01 mol/L磷酸氢二钾溶液,用20%磷酸溶液调节pH值至8.0)∶乙腈(20∶80),检测波长为210 nm,流速为1.0 mL/min,柱温为40℃。用该条件进行检测,加米霉素标准品在1.714～450.0μg/mL范围内呈良好的线性关系(R~2=0.999 7);重复性试验平均含量为98.18%,相对标准偏差(RSD)为0.85%;在高、中、低3个水平下的回收率为98%～102%,平均回收率为99.71%,RSD为0.94%;用确定的液相色谱检测条件对8批中试样品的含量进行测定,结果RSD为0.23%。说明该方法重复性好、准确性高,可用于加米霉素注射液含量的检测。     

不同方法测定反刍动物饲料NDF、ADF和木质素含量的比较

采用传统抽滤、ANKOM滤袋和CAU滤袋技术测定16种反刍动物饲料的中性洗涤纤维(NDF)、酸性洗涤纤维(ADF)和木质素(PL,高锰酸钾法)含量。结果表明:3种方法测定的大部分饲料的NDF、ADF和PL含量间没有差异(P>0.05),但在测定谷物性饲料NDF含量时,ANKOM滤袋技术测定值明显低于其他2种方法(P<0.05);在测定动物性蛋白质饲料时,CAU滤袋测定的NDF值明显高于其他2种方法(P<0.05)。3种方法测定的NDF、ADF和木质素含量均具有显著的线性相关关系(R2=0.990～0.996),其中2个滤袋技术测定值之间的相关程度最高(R2=0.992～0.996);2个滤袋技术测定值的变异系数(CV=2.20%～3.96%)均小于传统抽滤(CV=3.21%～4.91%)。     

Adverse effects of ketoconazole in dogs--a retrospective study

Although ketoconazole has been used extensively in dogs for the treatment of various fungal infections, information about adverse effects is mainly anecdotal. Common adverse effects in humans include dose-dependant anorexia, nausea and vomiting, allergic rashes and pruritus. Drug-induced hepatitis is very rare, but potentially fatal. The aim of this study was to evaluate the type and frequency of adverse effects associated with ketoconazole therapy in dogs treated for skin diseases and any possible influence of dosage, duration of therapy, signalment or concurrent medication. The medical records of 632 dogs treated with ketoconazole (2.6–33.4 mg/kg) were reviewed. Adverse effects occurred in 14.6% (92 dogs) and included vomiting (7.1%), anorexia (4.9%), lethargy (1.9%), diarrhea (1.1%), pruritus (0.6%), erythema (0.3%) and other adverse effects (2.5%). Of the dogs with other adverse effects, four of 16 (25%) were ataxic and three of these received concurrent ivermectin. Adverse effects were significantly more often recorded in dogs concurrently treated with ciclosporin ( P =  0.034) or ivermectin ( P =  0.007). Increased liver enzyme levels were reported rarely, and icterus was not seen in any of the dogs. However, monitoring liver enzymes during therapy is recommended, although this might not necessarily prevent severe idiosyncratic hepatotoxicity.     

The effects of ketoconazole and cimetidine on the pharmacokinetics of oral tramadol in greyhound dogs

Tramadol is administered to dogs for analgesia but has variability in its extent of absorption, which may hinder its efficacy. Additionally, the active opioid metabolite (M1) occurs in low concentrations. The purpose of this study was to determine if administration of oral tramadol with suspected metabolism inhibitors (ketoconazole, cimetidine) would lead to improved bioavailability of tramadol and M1. Six healthy Greyhounds were included. They were administered tramadol orally and intravenously, M1 intravenously, oral tramadol with oral ketoconazole and oral tramadol with oral cimetidine. Oral tramadol bioavailability was low (2.6%). Ketoconazole and cimetidine significantly increased tramadol bioavailability to 18.2% and 20.3%, respectively. The mean maximum plasma concentration of tramadol alone was 22.9 ng/ml, and increased to 109.9 and 143.2 ng/ml with ketoconazole and cimetidine, respectively. However, measured tramadol plasma concentrations were below the minimum concentration considered effective in humans (228 μg/ml). In all treatment groups, measured M1 concentrations (<7 μg/ml) were below concentrations associated with efficacy in humans. To conclude, tramadol and M1 concentrations were low and variable in dogs after oral dosing of tramadol, even in combination with cimetidine or ketoconazole, but effective concentrations in dogs have not been defined.     

Use of ketoconazole in the treatment of canine nasal aspergillosis

Fifteen dogs with nasal aspergillosis were treated with ketoconazole (5 mg/kg of body weight, q 12 h, PO) for 2 to 18 weeks. Four dogs whose conditions deteriorated during treatment received ketoconazole for less than the prescribed 6 weeks. Six months or more later, only 47% of the dogs were determined to be disease-free, on the basis of no fungal growth on culture. It was concluded that ketoconazole at this dosage is a useful treatment for canine nasal aspergillosis, but is no more effective than thiabendazole.     

The pharmacokinetics of ketoconazole and its effects on the pharmacokinetics of midazolam and fentanyl in dogs

KuKanich, B., Hubin, M. The pharmacokinetics of ketoconazole and its effects on the pharmacokinetics of midazolam and fentanyl in dogs. J. vet. Pharmacol. Therap . 33 , 42–49.
Ketoconazole inhibits the Cytochrome P450 3A12 (CYP3A12) metabolizing enzyme as well as the p-glycoprotein efflux pump. The extent and clinical consequence of these effects are poorly understood in dogs. The objective was to assess the pharmacokinetics of ketoconazole after single and multiple doses and the effect of multiple doses of ketoconazole on midazolam (a known CYP3A12 substrate) and the opioid fentanyl. Six greyhound dogs were studied. The study consisted of three phases. Phase 1 consisted of i.v. midazolam (0.23 mg/kg base) and fentanyl (15.71 μg/kg base). Phase 2 consisted of a single oral dose of ketoconazole (mean dose 12.34 mg/kg). Phase 3 consisted of i.v. midazolam (0.23 mg/kg) and fentanyl (10 μg/kg) after 5 days of oral ketoconazole (12.25 mg/kg/day). Ketoconazole significantly inhibited its own elimination with the mean residence time ( MRT ) increasing from 6.24 h in Phase 1 to 12.54 h in Phase 3. Ketoconazole significantly decreased the elimination of midazolam, as expected, with the MRT increasing from 0.81 to 1.49 h. The elimination of fentanyl was not significantly altered by co-administration of ketoconazole with the MRT being 3.90 and 6.35 h. The MRT was the most robust estimate of decreased drug elimination.     

Cyclosporin and ketoconazole interaction for treatment of perianal fistulas in the dog

OBJECTIVE: To assess the use of concurrent ketoconazole and low dose cyclosporin administration in a group of dogs with clinical evidence of perianal fistulas, and to determine if this combination could be used to manage perianal fistulas effectively. DESIGN: Prospective clinical trial PROCEDURE: Sixteen dogs with clinical evidence of perianal fistulas were given ketoconazole (10 mg/kg once daily) and cyclosporin (1 mg/kg twice daily initially) for 16 weeks. Blood cyclosporin assays were performed regularly and cyclosporin doses were altered to achieve a stable blood level above 200 ng/mL. Regular examinations assessed the dogs' general health, changes in clinical behaviour, fistula size and number. A complete blood count and serum biochemical analysis was performed in all dogs before and after the treatment period, and after 8 weeks of treatment in 12 dogs. Dogs were assessed for recurrence of lesions at 1, 3 and 12 months after the trial. RESULTS: All dogs showed marked improvement in lesions and behaviour within 14 days of the medication. Fourteen dogs completed the trial. Two dogs were excluded due to concurrent disease. Thirteen dogs (93%) showed complete resolution of fistulas during the treatment period. Seven dogs (50%) had no recurrence after 12 months. Recurrence was seen in three dogs (21%) at 8, 10 and 12 months after treatment, and in three dogs (21%) within 1 month of treatment. The medication was well tolerated. Side effects included transient anorexia, vomiting and lethargy in some dogs, increased shedding of hair and gingival hyperplasia. Ketoconazole administration allowed a dramatic reduction in cyclosporin dose (over 90% in 12 dogs and 80% in the other two) compared to previously reported cases treated with cyclosporin alone. CONCLUSION: The use of combined ketoconazole and cyclosporin provided an effective treatment for perianal fistulas. Outcomes were similar to those seen with cyclosporin alone, but allowed a significant reduction in cyclosporin dose and, therefore, cost. The use of immunosuppressive therapy in the treatment of perianal fistulas was effective and avoided many of the problems associated with surgical treament.     

Cyclosporine and ketoconazole for the treatment of perianal fistulas in dogs

OBJECTIVE: To evaluate efficacy and cost of using cyclosporine and ketoconazole for the treatment of perianal fistulas in dogs. DESIGN: Clinical trial. ANIMALS: 12 dogs with perianal fistulas. PROCEDURE: Dogs received cyclosporine and ketoconazole orally (target whole blood trough cyclosporine concentrations of 400 to 600 ng/ml). Study endpoints were resolution of clinical signs, remission, and recurrence of disease. Adverse effects and cost of medications were reported. Results were compared with those from previous studies in humans and in dogs in which single agent cyclosporine treatment for perianal fistulas was used. RESULTS: All dogs had resolution of clinical signs. Eight dogs went into remission; however, 5 of those 8 had recurrence of fistulas. Adverse effects of treatment were minimal and well tolerated. Cost of treatment was comparable to traditional surgical options and less than single agent cyclosporine treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of cyclosporine with ketoconazole is an effective and cost-comparable treatment for perianal fistulas in dogs.     

Treatment of canine blastomycosis with amphotericin B and ketoconazole

The treatment of 62 dogs with blastomycosis was reviewed to identify prognostic factors and response to various treatment regimens. Severity of lung involvement, as determined by radiography, and the number of nonsegmented neutrophils were useful prognostic factors. Females survived treatment better than did males, but females were more prone to relapse. Ketoconazole treatment at a dose of 10 mg/kg daily for 60 days was not as effective as amphotericin B. The most notable adverse effect of amphotericin B treatment was nephrotoxicosis . Treatment with amphotericin B followed by ketoconazole was as effective as amphotericin B alone and resulted in less nephrotoxicosis . Most dogs that had relapses were retreated effectively with amphotericin B and/or ketoconazole. Canine blastomycosis was shown to be a treatable disease, with a cure rate of approximately 75%.     

Effects of ketoconazole on the pharmacokinetics and pharmacodynamics of morphine in healthy Greyhounds

OBJECTIVE: To assess pharmacokinetics and pharmacodynamics of morphine and the effects of ketoconazole on the pharmacokinetics and pharmacodynamics of morphine in healthy Greyhounds. ANIMALS: 6 healthy Greyhounds, 3 male and 3 female. PROCEDURES: Morphine sulfate (0.5 mg/kg. IV) was administered to Greyhounds prior to and after 5 days of ketoconazole (12.7 +/- 0.6 mg/kg, PO) treatment. Plasma samples were obtained from blood samples that were collected at predetermined time points for measurement of morphine and ketoconazole concentrations by mass spectrometry. Pharmacokinetics of morphine were estimated by use of computer software. RESULTS: Pharmacodynamic effects of morphine in Greyhounds were similar to those of other studies in dogs and were similar between treatment groups. Morphine was rapidly eliminated with a half-life of 1.28 hours and a plasma clearance of 32.55 mL/min/kg. The volume of distribution was 3.6 L/kg. No significant differences in the pharmacokinetics of morphine were found after treatment with ketoconazole. Plasma concentrations of ketoconazole were high and persisted longer than expected in Greyhounds. CONCLUSIONS AND CLINICAL RELEVANCE: Ketoconazole had no significant effect on morphine pharmacokinetics, and the pharmacodynamics were similar between treatment groups. Plasma concentrations of ketoconazole were higher than expected and persisted longer than expected in Greyhounds.     

Comparative efficacies of oral ketoconazole and terbinafine for reducing Malassezia population sizes on the skin of Basset Hounds

The objective of this study was to compare the efficacy of oral ketoconazole and terbinafine for reducing population sizes of Malassezia yeasts on canine skin. Twenty-one Basset Hounds were randomised in three groups of seven according to Malassezia populations. Dogs in the first group were treated by oral administration of ketoconazole (Ketofungol) 200 mg, Janssen-Cilag) at 10 mg x kg-1, every 24 h with food, for 3 weeks. Dogs in the second group were treated by oral administration of terbinafine (Lamisil) 250 mg, Novartis) at 30 mg x kg-1, every 24 h with food, for 3 weeks. The seven remaining dogs were used as controls. Malassezia population sizes were assessed by use of contact plates on four cutaneous sites at days 7, 14 and 21. Both ketoconazole and terbinafine were effective in reducing the baseline levels of Malassezia organisms with no significant difference between the two drugs. In further studies, oral terbinafine should be evaluated for the management of canine cases of Malassezia dermatitis.     

Plasma cortisol response to ketoconazole administration in dogs with hyperadrenocorticism

The effect of orally administered ketoconazole on plasma cortisol concentration in dogs with hyperadrenocorticism was evaluated. Every 30 minutes from 0800 hours through 1600 hours and again at 1800 hours, 2000 hours, and 0800 hours the following morning, 15 clinically normal dogs and 49 dogs with hyperadrenocorticism had plasma samples obtained and analyzed for cortisol concentration. The mean (+/- SD) plasma cortisol concentration for the initial 8-hour testing period was highest in 18 dogs with adrenocortical tumor (5.3 +/- 1.6 micrograms/dl), lowest in 15 control dogs (1.3 +/- 0.5 micrograms/dl), and intermediate in 31 dogs with pituitary-dependent hyperadrenocorticism (PDH; 3.4 +/- 1.2 micrograms/dl). Results in each of the 2 groups of dogs with hyperadrenocorticism were significantly (P less than 0.05) different from results in control dogs, but not from each other. The same cortisol secretory experiment was performed, using 8 dogs with hyperadrenocorticism (5 with PDH; 3 with adrenocortical tumor) before and after administration at 0800 hours of 15 mg of ketoconazole/kg of body weight. Significant (P less than 0.05) decrease in the 8-hour mean plasma cortisol concentration (0.9 +/- 0.2 microgram/dl) was observed, with return to baseline plasma cortisol concentration 24 hours later. Twenty dogs with hyperadrenocorticism (11 with PDH, 9 with adrenocortical tumor) were treated with ketoconazole at a dosage of 15 mg/kg given every 12 hours for a half month to 12 months. The disease in 2 dogs with PDH failed to respond to treatment, but 18 dogs had complete resolution of clinical signs of hyperadrenocorticism and significant (P less than 0.05) reduction in plasma cortisol responsiveness to exogenous adrenocorticotropin (ACTH).(ABSTRACT TRUNCATED AT 250 WORDS)