苯丙醇羟丙基-β-环糊精包合片的生物等效性研究

Study on Bioequivalence of Phenylpropanol Tablets in Beagle Dogs

目的评价苯丙醇羟丙基-β-环糊精包合片与苯丙醇胶丸的生物等效性。方法采用双制剂双周期交叉实验设计,8只Beagle犬随机分为两组,分别单剂量口服苯丙醇-羟丙基-β-环糊精包合片(受试制剂)和苯丙醇胶丸(参比制剂)各100mg(以苯丙醇计),12d后交叉服药。于给药后0.083,0.25,0.5,1.0,1.5,2,3,4,6,8,12,14,24,36h取血样,0.5mL血浆加入15%高氯酸溶液100μL,涡旋振荡40s,4000r.min-1离心10min,上清液经0.45μm滤膜过滤后,取20μL进行HPLC分析。流动相为甲醇-双蒸水=60:40,流速为1.0mL.min-1,色谱柱为Diamond C18柱(4.6mm×250mm,5μm),柱温30℃。紫外检测波长为210nm。结果犬血浆中杂质不干扰样品的测定,犬血浆中苯丙醇质量浓度在0.01287～25.73mg.L-1内线性关系好(r=0.99999);最低检测质量浓度为0.005mg.L-1(以S/N≥3计)。参比制剂的t1/2为(5.68±1.11)h,tmax和ρmax分别为(0.8±0.5)h和(4.46±0.73)mg.L-1;受试制剂t1/2为(4.53±0.72)h,tmax和ρmax分别为(1.0±0.7)h和(3.05±0.44)mg.L-1,其相对生物利用度为(104.9±14.0)%。结论所建立方法准确可靠,符合生物样品分析要求;犬单剂量口服受试制剂与参比制剂后,药动学统计数据表明,两种制剂生物等效。

OBJECTIVE To develop a HPLC assay for determining phenylpropanol in dog plasma and to evaluate the pharmacokinetics and bioequivalence of two phenylpropanol preparations in beagle dogs. METHODS According to the crossover experimental design, the two preparations （the reference and test preparations ） were tested within 12 d interval of two cycle, eight beagle dogs were grouped into two groups. A single dose of 100 mg reference preparations （phenylpropanol soft gelatin capsule） and test preparations （phenylpropanol tablet clathrated by cyclodextrin） was administrated to the two groups respectively. The plasma samples were gathered at 0. 083,0. 25,0. 5,1.0,1.5,2,3,4,6,8,12,14,24 and 36 h after dosage, and then being deproteined by 15% perchloric acid solution. The supematant was filtered through a 0. 45μm filter membrane, then 20μL filtrate was separated on a C18 reversed phase column with a mobile phase of methanol-water（60： 40） ,and the flow rate was at 1.0 mL · min^-1. The column remained at 30 % and phenylpropanol was detected at 210 nm. RESULTS Calibration curve obtained was linear over the range of 0. 012 87 - 25.73 mg· L^-1 （ r = 0. 999 99） and the detective limit was 0. 005 mg· L^-1 （S/N≥3）. t（1/2）, tmax and ρmax were （5.68 ±1.11 ）h, （0. 8±0.5）h, （4. 46± 0. 73 ） μg ·L^ -1 for the reference preparations and （4. 53 ± 0. 72 ） h, （ 1.0 ± 0. 7 ） h, （ 3.05 ± 0.44 ） μg ·L^ -1 for the test preparations. The relative bioavalability of the test preparations was （ 104. 9 ± 14. 0 ） %. The results of variance analysis and two one-sided t-test showed that there was no significant difference between the two preparations in the AUC and tmax But the ρmax was different. CONCLUSION The assay was proved to be sensitive, accurate and convenient. The two preparations were bioequivalent .