摘要
本课题首先研究了0.5%复方伊维菌素注射液的制备方法,建立了复方伊维菌素注射液的含量测定方法并对其进行了稳定性考察。建立了猪血浆中伊维菌素和盐酸氧阿苯达唑的提取方法及反相高效液相色谱(RP-HPLC)检测法。使用Agilent 1100型高效液相色谱(HPLC)仪,Agilent TC-C_(18)(5μm,4.6×150mm,I.D.),保护柱为Agilent TC-C_(18)(5μm,4.6×12.5mm,I.D.);色谱条件为两种:色谱条件Ⅰ(测定复方伊维菌素注射液中的伊维菌素),流动相为甲醇:水=(90:10,v/v);流速:1.0mL/mim进样量:20μL:柱温:30℃;检测波长:245nm。色谱条件Ⅱ(测定复方伊维菌素注射液中的盐酸氧阿苯达唑),流动相为乙腈:甲醇:水=(13:15:72,v/v/v),含1.25%三乙胺,磷酸调节pH至3.1;流速:1.0mL/min:进样量:20μL:柱温:40℃;检测波长:290nm。对0.5%复方伊维菌素注射液经过单次肌内注射给药以后在猪体内的药物动力学进行了研究,所选健康猪(n=6)体重为28.94±2.67kg,给药剂量为0.3mg/kg b.w.。结果表明,伊维菌素药时曲线符合有吸收一室开放模型,动力学方程为:C=45.19(e~(-0.30t)-e~(-1.56t))。主要药物动力学参数为:吸收速率常数K_a 1.56±0.10d~(-1);消除半衰期t_(1/2ke) 2.30±0.16d;药时曲线下面积AUC 120.86±6.98ng·mL~(-1)·d;表观分布容积V_d/F 8.22±0.12L/kg:清除率CL_b 2.49±0.15L·d~(-1)·kg(-1);达峰时间T_p 1.31±0.079d:达峰浓度C_(max) 24.57±0.15ng/mL:盐酸氧阿苯达唑药时曲线符合有吸收二室开放模型,动力学方程为:C=2.21 e~(-0.39t)+7.73e~(-0.19t)-9.94 e~(-0.43t)。其主要药物动力学参数为:吸收速率常数k_a 0.43±0.09h~(-1);消除半衰期t_(1/2β)3.85±0.70h:药时曲线下面积AUC 24.37±3.62μg·mL~(-1)·h:表观分布容积V_d/F 1.18±0.17L/kg;清除率CL_b 0.21±0.03L·h~(-1)·kg~(-1);达峰时间T_p 3.63±0.64h:达峰浓度C_(max) 2.33±0.37μg/mL。
通过药物动力学试验可知,以0.3mg/kg b.w.剂量单次肌内注射给药后,伊维菌素在猪体内吸收速度较快,分布广,消除缓慢;盐酸氧阿苯达唑在猪体内吸收速度较快,分布广,消除较快。
In this research,preparing compound ivermectin injection,determining the content of principal agents in compound ivermectin injection and studying its stability were all performed.The research has established the method of detection ivermectin(IVM) and albendazole sulphoxide hydrochloride(ABZSO) after extraction from plasma of pigs, using reversed-phase high performance liquid chromatography(RP-HPLC).Separation was achieved on an Agilent TC-C_(18) reversed-phase analytical column(5μm,6×150mm, I.D.).An Agilent TC-C_(18) guard column(5μm,4.6×12.5mm,I.D.) was used to protect the analytical column.The mobile phase I for the analysis of IVM was methanol:water (90:10,v/v),at a flow rate of 1.0 mL/min,the injection volume was 20μL,elution was performed at an oven temperature of 30℃and IVM was determined by HPLC with UV detection at 245 nm.The mobile phaseⅡfor the analysis of ABZSO was a solution of 1.25%triethylamine in acetonitrile:methanol:water(13:15:72,v/v/v),pH was adjusted to 3.1 using 85%o-phosphoric acid,at a flow rate of 1.0 mL/min,the injection volume was 20μL,elution was performed at an oven temperature of 40℃and IVM was determined by HPLC with UV detection at 290nm.Pharmacokinetics of IVM and ABZSO were investigated,following a single intramuscular administration at dose of 0.3mg/kg b.w. (28.94±2.67kg) in healthy pigs(n=6).The result showed that IVM concentration-time data was fitted to a one-compartment model with first order absorption.The absorption rate constant(K_a) of IVM was calculated to be 1.56±0.10d~(-1),and the elimination half-life (t1/2ke) of the drug was 2.30±0.16d.The area under the serum concentration-time curve (AUC) was 120.86±6.98ng.mL~(-1)·d.The distribution volume(V_d/F) of lVM was computed to be 8.22±0.12L/kg.The total clearance of IVM(CL_b) was estimated to be 2.49±0.15L·d~(-1)·kg~(-1) and the maximal plasma concentration(C_(max)) was calculated as 1.31±0.079d and 24.57±0.15ng/mL.Besides,the result showed that ABZSO concentration-time data was fitted to a two-compartment model with first order absorption.The absorption rate constant(K_a) of ABZSO was calculated to be K_a 0.43±0.09h~(-1),and the elimination half-life(t1/2β) of the drug was 3.85±0.70h.The area under the serum concentration-time curve(AUC) was 24.37+3.62μg.mL~(-1)·h.The distribution volume(V_a/F) of ABZSO.was computed to be 1.18±0.17L/kg.The total clearance of ABZSO(CL_b) was estimated to be 0.21±0.03L·h~(-1)·kg~(-1).The time-point of maximal plasma concentration of the drug(T_p) and the maximal plasma concentration (C_(max)) were calculated as 3.63±0.64h and 2.33±0.37μg/mL.
The results of present studies revealed that IVM was absorbed fast,distributed extensively and eliminated slowly and ABZSO was absorbed fast,distributed extensively and eliminated fast in pigs.
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