地昔尼尔乳剂的制备及其HPLC测定方法的建立

Preparation and establishment of HPLC determination of emulsion of dicyclanil

为制备地昔尼尔乳剂并建立其质量浓度的测定方法,以稳定常数和离心稳定性作为乳剂稳定性评价指标,采用单因素试验和正交设计试验法考察处方因素和工艺因素对乳剂稳定性的影响,以筛选出乳剂的最优处方和制备工艺;采用高效液相色谱法(HPLC)测定乳剂的药物质量浓度。结果显示,地昔尼尔乳剂的最优处方为地昔尼尔50g/L、乳化剂70mL/L、油相125mL/L、助乳化剂160mL/L、稳定剂为3.5g/L;地昔尼尔乳剂的制备方法为乳化温度50℃,20 000r/min高速搅拌10min。建立的HPLC色谱条件为色谱柱X Bridge C18(4.6mm×250mm,5μm),流动相乙腈与水(6mL/L三乙胺+2mL/L甲酸)的体积比为10∶90,流速0.8mL/min,柱温30℃,紫外检测波长267nm,进样体积20μL。在10~100μg/mL的质量浓度范围内,峰面积与药物浓度呈良好线性关系(r2=0.999 9,n=7),回归方程为y=148.01x+91.629。精密度试验相对标准偏差均小于2%,在80%、100%和120%3个添加水平时,平均回收率为98.36%,精密度、回收率均符合要求。定量限和检测限分别为10ng/mL和20ng/mL,测得3批地昔尼尔乳剂的平均质量浓度为49.29mg/mL,为标示量的98.57%。结果表明,该处方合理、制备工艺可靠,所建立的高效液相色谱法可用于地昔尼尔乳剂的检测。

To develop an emulsion of dicyclanil,and to establish a method for the determination of the emulsion,the centrifugal stability constant and the stability parameter Kewere adopted as the coindicator.Single factor tests and orthogonal designing method were used to investigate the effection of formulation and technological factors to emulsion stability.The results showed that the optimal formulation was composed by dicyclanil 50g/L,emulsifier 70mL/L,Aethylis oleas 125mL/L,assistant emulsifier 160mL/L and stabilizer 3.5g/L.Technological conditions were optimized,emulsifying temperature(50℃),and high shear speed(20 000r/min for 10min).The content of the emulsion was determined by using high performance liquid chromatography.An C18column(4.6mm×250mm,5μm)was used,and the proportion of acetonitrice and water which was added with 2mL/L methane acid and 6mL/L triethanolamine was 10∶90by the mobile phase.Flow rate was 0.8mL/min,and the column temperature was 30℃,and the detection wavelength was 267 nm,with injection volume of 20μL.Specificity test showed that dicyclanil was undisturbed in the peak position.The linear range of the calibration curve for dicyclanil was from 10 to 100μg/mL(r2=0.999 9,n=7),taking concentration as the abscissa and peak area for the vertical axis linear regression,and regression equation showed y=148.01x+91.629.The relative standard deviation of precision were less than 2%.Prepare nine blank accessories and then add for the equivalent concentration of the test emulsion of dicyclanil reference 80%,100%and 120%,three copies of each.At the spiked levels of80%,100% and 120%,the average recovery rate was 98.36%.The dicyclanil was gradually diluted with mobile phase,according to the signal to noise ratio(SNR)3∶1counted as the limit of detection(LOD),and according to SNR10∶1calculated as the limit of quantification(LOQ).The LOQand LOD were 10ng/mL and 20ng/mL.Three batches of emulsion were diluted with mobile phase,filtered by the organic membrane,and then inject to detection.The content of dicyclanil was changed to the indicated value content of the sample.The average signal content of the emulsion was 98.57%.The prescription was reasonable and the preparation technology was reliable.The determination method has high accuracy and sensitivity and can be used for emulsion of dicyclanil test.