伊维菌素微乳中伊维菌素的含量测定方法研究

为建立伊维菌素微乳中伊维菌素含量的高效液相色谱（HPLC）测定方法,选用Hypersil ODS2 （5 μm,4.6 mm×250 mm）色谱柱,流动相为甲醇∶乙腈∶水为35∶60∶5（V/V/V）,检测波长为244 nm,柱温为30 ℃,流速为1 mL/min进行测定。结果显示,伊维菌素在该色谱条件下,系统适应性良好,在80~320 μg/mL浓度范围内线性关系良好,回归方程为：Y=22 700X+2 510,R²=0.9998,总平均回收率为101.90%±2.94%,RSD为2.88%,对中试生产的3批伊维菌素微乳进行含量测定,RSD为1.86%。表明该含量测定方法准确可靠,重现性好,可用于伊维菌素微乳中伊维菌素含量的测定,并为该新型制剂的质量标准的制定和质量评价提供依据,也为后期的临床安全应用提供可靠的参考。     

RP-HPLC检测复方注射剂中伊维菌素和吡喹酮的含量

为了用高效液相色谱法同时测定复方注射剂中伊维菌素和吡喹酮的含量。采用Hyper-C lone 5u C18柱（250 mm×4.60 mm,5μm）,以乙腈-甲醇-水（53∶35∶12）为流动相,流速为1.0mL/m in,检测波长254 nm,柱温30℃的液相检测方法。结果表明,在此色谱条件下,吡喹酮和伊维菌素的分离良好,吡喹酮和伊维菌素分别在30~500μg/mL和5~40μg/mL浓度范围内峰面积与浓度呈良好的线性关系（r2〉0.99）。按外标法以峰面积计算进行测定,吡喹酮和伊维菌素的平均回收率分别为98.93%和95.89%,RSD分别为0.33%和1.58%（n=9）。说明该方法简便、快速、准确,适用于复方吡喹酮注射液中吡喹酮和伊维菌素含量的测定。     

伊维菌素长效透皮剂和普通注射剂在家兔体内的药代动力学比较研究

为比较研究制备的伊维菌素长效透皮制剂与普通伊维菌素注射剂药物代谢及药效时间,本研究制备伊维菌素含量分别为0.5%、1.0%和1.5%的长效透皮制剂,采用高效液相色谱法检测不同药量相同体积伊维菌素长效透皮制剂和普通伊维菌素注射剂（1.0%）在家兔体内的药代动力学,并通过PKSolver药代动力学处理软件对数据进行分析。结果显示,0.5%、1.0%、1.5%伊维菌素长效透皮剂和1.0%普通注射剂吸收半衰期分别为0.81、0.52、1.02和0.12 d;达峰时间为1.55、0.97、1.62和0.42 d;峰浓度为47.36、72.02、115.30和99.53 ng/mL;消除半衰期为3.61、5.92、5.59和1.79 d;平均滞留时间为5.27、7.37、5.13和2.16 d;药时曲线面积为1 488.70、3 081.98、3 161.20和480.00 ng·d/mL,伊维菌素长效透皮剂体内维持有效药物浓度的时间长达35 d,普通注射剂仅为9 d。结果表明,伊维菌素长效透皮剂效果稳定,可进行更深入的研究。     

伊维菌素聚氰基丙烯酸丁酯纳米粒的制备

以BCA为单体材料,采用乳液聚合法制备IVM-PBCA-NP纳米粒,经药物吸附和PVP包被后形成PVP-IVM-PBCA-NP载药纳米粒。经正交试验设计确定以BCA:IVM投量为4:1(g/g),BCA投量为0.4:25(V/V),乳化剂F68浓度为2%,搅拌速度600r/min,反应体系pH值为2、3、6,反应时间3h,为最佳制备条件。在该条件下所制备的PVP-IVM-PBCA-NP药物包封率为(45.43±1.65)%,载药量(9.95±0.23)%,主要粒径为296nm。     

伊维菌素微乳制剂的过敏性研究

本试验旨在观察伊维菌素微乳制剂对动物的过敏性反应。以《化学药物刺激性、过敏性和溶血性研究技术指导原则》为指导,选用豚鼠及Wistar大鼠为试验动物,采用全身主动过敏试验和被动皮肤过敏试验观察伊维菌素微乳制剂对动物的致敏作用。结果显示伊维菌素微乳制剂对豚鼠和Wistar大鼠的过敏性反应均为阴性。本试验为伊维菌素微乳制剂的安全性评价提供了试验依据。     

伊维菌素微乳无菌检查方法学的研究

本试验旨在对伊维菌素微乳制剂进行无菌检查方法学验证和无菌检查试验,确认本试验所用的方法适用于该制剂的无菌检查。按《中国兽药典》2005版一部(附录118)所载"无菌检查法"项下进行试验,通过对阳性对照菌、不同量冲洗液等条件的选择,采用薄膜过滤法对10瓶供试品(每种试验菌的样品量)进行检测,建立了无菌检查方法。经方法验证,用400mL 0.1%蛋白胨水溶液冲洗后,含供试品容器中的7个阳性菌试验组与阳性菌对照组相比均生长良好,说明供试品的该检验量在该检验条件下无抑菌作用或其抑菌作用可以忽略不计,可以用该方法进行供试品的无菌检查。对3批供试品进行无菌检查,阳性对照菌均在24h内生长良好,阴性对照均澄清,无菌生长,3批供试品均澄清,无菌生长,无菌检查试验结果符合规定。     

Study on Sterility Test Method of Ivermectin Microemulsion

The assay was aimed to study the sterility test and validation test of ivermectin microemulsion preparation and establish a sterility test method for ivermectin microemulsion preparation.The test method was carried out according to the method in volumeⅠ, Chinese Veterinary Pharmacopoeia Edition 2005.By choosing positive control bacteria and defining washing volumes in sterility test, the membrane-filter method was used to test the quantity of 10 bottles of test samples, and the sterility test was established.The result of method validation test showed that the test and all of positive control bacteria and microorganism growth after each filter being washed with 400 mL 0.1% peptone solution.It illustrated that the samples had no antimicrobial activity under the sample quantity and test condition.This method was available for sterility test of ivermectin microemulsion preparation.Using this method to test three lots test samples, the results showed that the positive control bacteria grew well within 24 h.The negative control bacteria and three lots test samples were sterile.It indicated that sterility test results met the requirements.     

联合应用阿苯达唑和伊维菌素在线虫感染鄂尔多斯细毛羊体内的药动学研究

为阐明联合应用阿苯达唑(ABZ)和伊维菌素(IVM)在胃肠道线虫感染鄂尔多斯细毛羊体内的药动学互作关系,以感染胃肠道线虫的鄂尔多斯细毛羊为研究对象,比较研究了单独或联合应用阿苯达唑和伊维菌素后的药物动力学特征。通过粪便虫卵检查法,选取感染胃肠道线虫的鄂尔多斯细毛羊15只,随机分成3组,每组5只。第1组口服给予阿苯达唑(15mg/kg),第2组皮下注射伊维菌素(0.2mg/kg),第3组皮下注射伊维菌素(0.2mg/kg)的同时口服阿苯达唑(15mg/kg)。于给药后不同时间,由颈静脉采集血样,分离血浆,并用高效液相色谱法测定各时间点血浆阿苯达唑、阿苯达唑亚砜、阿苯达唑砜和伊维菌素浓度,并用PK Solution 2.0药物动力学软件计算出各药动学参数。结果表明,联合用药组绵羊血浆伊维菌素峰浓度(Cmax)、药时曲线下面积(AUC)和平均滞留时间(MRT)分别为44.80ng/mL±6.12ng/mL、5 007.46ng.h/mL±1 301.42ng.h/mL和85.47h±5.03h,均显著(P<0.05)小于单独用药组的对应参数值67.62ng/mL±9.06ng/mL、7 125.08ng.h/mL±908.52ng.h/mL和113.39h±9.00h。口服阿苯达唑组绵羊血浆中仅检测到了阿苯达唑砜和阿苯达唑亚砜,而未检测到阿苯达唑母药。联合用药后,除阿苯达唑砜的达峰时间(T max)显著推迟外,阿苯达唑砜和阿苯达唑亚砜的其他各参数之间均无显著性差异。因此,联合应用IVM和ABZ可影响它们在胃肠道线虫感染鄂尔多斯细毛羊体内的药动学特征,且对伊维菌素药动学特征的影响尤为明显,在临床联合用药过程中应予以重视。     

伊维菌素脂质体的制备及质量控制研究

为研究伊维菌素脂质体 ,以卵磷脂和胆固醇为载体 ,采用注入法制备了伊维菌素脂质体 ,并应用紫外倍率系数法测定脂质体中伊维菌素的含量 ,采用高速离心法测定其包封率。结果表明 ,采用注入法制备的伊维菌素脂质体稳定性好 ,大小均匀 ,包封率效果良好 (77.42 %以上 ) ,紫外倍率系数法能消除辅料干扰 ,可作为伊维菌素脂质体质量控制的有效方法     

Plasma dispositions and concentrations of ivermectin in eggs following treatment of laying hens

AIMS: To determine the plasma disposition and concentrations of ivermectin (IVM) in eggs produced by laying hens following S/C, oral and I/V administration.

METHODS: Twenty-four laying hens, aged 37 weeks and weighing 1.73 (SD 0.12) kg were allocated to three groups of eight birds. The injectable formulation of IVM was administered either orally, S/C, or I/V, at a dose of 0.2?mg/kg liveweight, following dilution (1:5, v/v) with propylene glycol. Heparinised blood samples were collected at various times between 0.25 hours and 20 days after drug administration. Eggs produced by hens were also collected daily throughout the study period. Samples of plasma and homogenised egg were analysed using HPLC.

RESULTS: Maximum concentrations of IVM in plasma and mean residence time of IVM were lower after oral (10.2 (SD 7.2) ng/mL and 0.38 (SD 0.14) days, respectively) than after S/C (82.9 (SD 12.4) ng/mL and 1.05 (SD 0.24) days, respectively) administration (p<0.01). The time to maximum concentration and elimination half-life were shorter following oral (0.14 (SD 0.04) and 0.23 (SD 0.11) days, respectively) than S/C (0.25 (SD 0.00) and 1.45 (SD 0.45) days, respectively) administration (p<0.01). IVM was first detected in eggs 2 days after treatment in all groups and was detected until 8 days after oral and I/V administration, and until 15 days after S/C administration. Peak concentrations of IVM were 15.7, 23.3 and 1.9?µg/kg, observed 2, 5 and 4 days after I/V, S/C and oral administration, respectively.

CONCLUSIONS AND CLINICAL RELEVANCE: The low plasma bioavailability of IVM observed after oral administration in laying hens could result in lower efficacy or subtherapeutic plasma concentrations, which may promote the development of parasitic drug resistance. Due to high IVM residues in eggs compared to the maximum residue limits for other food-producing animal species, a withdrawal period should be necessary for eggs after IVM treatment in laying hens.     

5种伊维菌素注射液在家兔体内的比较药动学研究

试验旨在研究5种不同处方的伊维菌素注射液在家兔体内的比较药动学特征。50只健康家兔随机分为5组,通过单次给药剂量(5 mg/kg体重)皮下注射伊维菌素注射液,3P97药动学计算软件处理血浆药物浓度—时间数据。家兔皮下注射伊维菌素注射液属于一级吸收一室开放模型。自制伊维菌素注射液的:T_1/2(Ka)(8.10±3.68) h、T_1/2(Ke)(36.34±14.63) h、T_max (22.38±8.63) h、 C_max (354.01±57.75) ng/mL、 AUC (25843±3029) ng/(mL·h)。经SPSS统计学分析软件分析显示自制伊维菌素注射液的T_1/2(Ka)、T_1/2(Ke) 无显著性差异,但C_max 分析显示与C、D存在显著性差异(P<0.05),与A、B无显著性差异(P>0.05),但明显高于A、B的C_max。AUC平均值明显高于其余4种,说明自制伊维菌素注射液在家兔体内具有吸收好,消除快,生物利用度高等优点,临床可安全使用,具有更好的抗寄生虫作用。     

乳腺康注射液质量标准研究

本研究旨在建立乳腺康注射液质量标准。按照《中国药典》（2015版）相关要求,分别建立乳腺康注射液中甘草、青皮、蒲公英的薄层色谱定性鉴别和重金属的检查方法并规定限度,采用HPLC法建立甘草苷、甘草酸铵、橙皮苷及咖啡酸的含量测定方法并规定限度。结果表明,甘草、青皮、蒲公英的定性鉴别专属性强;甘草苷在0.0459~0.7340 μg,甘草酸铵在0.0661~1.0580 μg,橙皮苷在0.0592~0.9470 μg,咖啡酸在0.0384~0.6140 μg浓度与峰面积呈良好的线性关系;相关系数r分别为0.997、0.9990、0.9999、1.000;甘草苷、甘草酸铵、橙皮苷及咖啡酸的平均加样回收率分别为101.25%、99.87%、98.39%、103.49%,相对标准偏差（RSD）值分别为4.82%、3.63%、2.94%、0.85%;乳腺康注射液中含甘草苷、甘草酸铵、橙皮苷及咖啡酸分别不低于88.90、138.48、136.50、21.54 μg/mL;按乳腺康注射液每日最大使用剂量计算,铅、镉、砷、汞、铜分别不得超过23、3、6、2及150 μg。本研究建立的质量标准可为控制乳腺康注射液的质量提供参考。     

The Effect of Ivermectin on Convulsions in Rats Produced by Lidocaine and Strychnine

Ivermectin is one of the most commonly used drugs in pharmacotherapy of parasitic diseases in domestic and wild animals caused by parasitic nematodes and arthropods. However, ivermectin and other avermectins very often produce side-effects in hosts. The most dominant clinical symptom of ivermectin toxicity in domestic and wild animals is CNS depression. In nematodes, the target site of ivermectin’s action is glutamate-gated chloride-channel receptor and GABA receptor. The depressive effect of ivermectin in mammals might include more than one mechanism; therefore, the anticonvulsive effect of ivermectin against convulsions caused by lidocaine and strychnine was evaluated. Ivermectin antagonized lidocaine- and strychnine-induced convulsions in rats, although these have different mechanisms. In the present study, the anticonvulsive ED₅₀ of ivermectin for lidocaine-induced convulsions was 2.44 mg/kg (95% CL 1.67 to 3.57 mg/kg), whereas for convulsions induced by strychnine it was higher at 4.25 mg/kg (95% CL 2.32 to 3.78 mg/kg). At the same time, both anticonvulsive doses are significantly lower then the observed LD₅₀ of ivermectin (18.20 mg/kg). Furthermore, flumazenil (0.1 and 0.2 mg/kg), an antagonist of benzodiazepine receptors, antagonizes just one part of these anticonvulsive effects of ivermectin. Our results show the significant anticonvulsive properties of ivermectin and support the findings that ivermectin in the CNS of mammals produces multiple inhibitory effects, probably through participation in the function of GABA-sensitive and GABA-insensitive chloride channels.     

Study on Quality Standard of Ruxiankang Injection

The aim of this study was to establish the quality standard of Ruxiankang (RXK) injection. According to the relevant requirements of the Chinese Pharmacopoeia (2015 edition), TLC identification,related substances examination and heavy metal examination method of RXK injection were established and the limits were determined. HPLC method was used to establish the content determination method of liquiritin, ammonium glycyrrhizinate, hesperidin and caffeic acid, and determined the limit. The specificity of Glycyrrhizae Radix et Rhizoma, Citri Reticulatae Pericarpium and Taraxaci Herba was strong. Liquiritin in 0.0459 to 0.7340 μg, ammonium glycyrrhizate in 0.0661 to 1.0580 μg, hesperidin in 0.0592 to 0.9470 μg, caffeic acid in the 0.0384 to 0.6140 μg concentration showed a good linear relationship with peak area. The correlation coefficients were 0.997,0.9990,0.9999 and 1.000,respectively. The average recoveries were 101.25%, 99.87%, 98.39% and 103.49%,respectively. The RSD values were 4.82%, 3.63%, 2.94% and 0.85%,respectively. The contents of liquiritin, ammonium glycyrrhizinate, hesperidin and caffeic acid of RXK injection were not less than 88.90, 138.48, 136.50 and 21.54 μg/mL,respectively. According to the maximum daily dose of RXK injection, Pb,Cd, As, Hg and Cu should not exceed 23,3,6,2 and 150 μg, respectively. The quality standard established in this study could provide reference for controlling the quality of RXK injection.     

伊维菌素预混剂质量标准中鉴别方法的商榷

为完善伊维菌素预混剂质量标准,对伊维菌素预混剂按质量标准中的薄层色谱鉴别方法进行了研究。结果发现：该鉴别方法存在缺陷,不能正确评价产品的质量。经过实验研究分析,对该标准提出修改意见,以使更加科学客观,可操作性更强。     

伊维菌素吡喹酮咀嚼片溶出度方法的建立

为了建立同时测定伊维菌素吡喹酮咀嚼片两种主药溶出度的测定方法,采用高效液相色谱法测定在不同pH值溶出介质、不同浓度表面活性剂和不同转速的条件下伊维菌素吡喹酮咀嚼片两种主药的溶出曲线,确定了伊维菌素吡喹酮咀嚼片的溶出方法。结果显示,三批样品45 min时伊维菌素、吡喹酮两种主药的溶出度均大于85%。建立的溶出度测定方法可用于伊维菌素吡喹酮咀嚼片溶出度的测定。     

伊维菌素聚酯酸酐微球的制备

将2种聚酯酸酐（P（SA：RA20：80）,P（SA：RA30：70））分别与伊维菌素溶于三氯甲烷中作为有机相,以聚乙烯醇水溶液为水相,用乳化溶剂挥发法制备出2种伊维菌素聚酯酸酐微球。采用光学显微镜考察所制备微球的形态及粒径,紫外分光光度法测定载药量和包封率。结果显示,制备出的P（SA：RA30：70）／IVM微球的平均粒径为（72．240±24．747）μm,载药量为19．67％,包封率为87．70％。P（SA：RA20：80）/IVM微球的平均粒径为（64．18±26．14）μm,载药量为17．72％,包封率为87．27％。这表明采用乳化溶剂挥发法成功制备出2种伊维菌紊聚酯酸酐微球。     

伊维菌素纳米乳注射液的研制与质量安全性评价

本研究旨在研制伊维菌素纳米乳注射液(5%)并对其理化性能、载药量、稳定性和急性毒性进行评价。采用伪三元相图法进行处方筛选,在常温下,以黏度、电导率、折光率、Z电位、平均粒径为指标考察纳米乳的理化性能;在不同的温度、光照和湿度条件下以药物含量为指标考察纳米乳的稳定性;采用HPLC法测定载药量,用小鼠灌胃的方法进行急性毒性研究。结果表明,该纳米乳在透射电镜下观察其乳滴呈球状,平均粒径为70 nm;不同温度、光照和湿度条件下含量无明显变化,稳定性良好;小鼠急性毒性试验表明该纳米乳注射液(0.1%)属于实际无毒。结果提示,本研究研制的伊维菌素纳米乳注射液(5%)制备简单,是一种质量稳定、安全性高的新制剂。     

Assessment of the long‐acting ivermectin formulation in sheep: Further insight into potential pharmacokinetic interactions

The aim of the current study was to evaluate the in vivo pharmacokinetic of ivermectin (IVM) after the administration of a long‐acting (LA) formulation to sheep and its impact on potential drug‐drug interactions. The work included the evaluation of the comparative plasma profiles of IVM administered at a single therapeutic dose (200 μg/kg) and as LA formulation at 630 μg/kg. Additionally, IVM was measured in different gastrointestinal tissues at 15 days posttreatment with both IVM formulations. The impact of the long‐lasting and enhanced IVM exposure on the disposition kinetics of abamectin (ABM) was also assessed. Plasma (IVM and ABM) and gastrointestinal (IVM) concentrations were analyzed by HPLC with fluorescent detection. In plasma, the calculated C_max and AUC_0‐t values of the IVM‐LA formulation were 1.47‐ and 3.35‐fold higher compared with IVM 1% formulation, respectively. The T_1/2ab and T_max collected after administration of the LA formulation were 2‐ and 3.5‐fold longer than those observed after administration of IVM 1% formulation, respectively. Significantly higher IVM concentrations were measured in the intestine mucosal tissues and luminal contents with the LA formulation, and in the liver, the increase was 7‐fold higher than conventional formulation. There was no drug interaction between IVM and ABM after the single administration of ABM at 15 days post‐administration of the IVM LA formulation. The characterization of the kinetic behavior of the LA formulation to sheep and its potential influence on drug‐drug interactions is a further contribution to the field.     

Ivermectin disposition kinetics after subcutaneous and intramuscular administration of an oil-based formulation to cattle.

Slight differences in formulation may change the plasma kinetics and ecto-endoparasiticide activity of endectocide compounds. This work reports on the disposition kinetics and plasma availability of ivermectin (IVM) after subcutaneous (SC) and intramuscular (IM) administration as an oil-based formulation to cattle. Parasite-free Aberdeen Angus calves (n = 24; 240-280 kg) were divided into three groups (n = 8) and treated (200 microg/kg) with either an IVM oil-based pharmaceutical preparation (IVM-TEST formulation) (Bayer Argentina S.A.) given by subcutaneous (Group A) and intramuscular (Group B) injections or the IVM-CONTROL (non-aqueous formulation) (Ivomec, MSD Agvet) subcutaneously administered (Group C). Blood samples were taken over 35 days post-treatment and the recovered plasma was extracted and analyzed by HPLC using fluorescence detection. IVM was detected in plasma between 12 h and 35 days post-administration of IVM-TEST (SC and IM injections) and IVM-CONTROL formulations. Prolonged IVM absorption half-life (p < 0.05) and delayed peak plasma concentration (p < 0.001) were obtained following the SC administration of the IVM-TEST compared to the IVM-CONTROL formulation. No differences in total plasma availability were observed among treatments. However, the plasma residence time and elimination half-life of IVM were significantly longer after injection of the IVM-TEST formulation. IVM plasma concentrations were above 0.5 ng/ml for 20.6 (CONTROL) and 27.5 days (IVM-TEST SC), respectively (p < 0.05). The modified kinetic behaviour of IVM obtained after the administration of the novel oil-based formulation examined in this trial, compared to the standard preparation, may positively impact on its strategic use in cattle.