氟苯尼考纳米乳含量测定方法的建立及稳定性研究
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摘要
为了建立高效液相色谱法测定氟苯尼考纳米乳中氟苯尼考含量的方法并对其稳定性进行研究,采用Iner Sustain C18(5μm,4.6 mm×250mm)色谱柱,以乙腈-水-磷酸(35∶65∶0.02,V/V/V)为流动相,流速1.0 m L/min,检测波长224 nm,进样量20μL,柱温25℃,考察方法的系统适用性、线性、精密度、加样回收率等,并通过影响因素试验及加速试验考察氟苯尼考纳米乳的稳定性。结果显示:氟苯尼考纳米乳在该色谱条件下,系统适应性良好,在4.12~67.74μg/m L浓度范围内线性关系良好,线性回归方程为:A=49 295C+2 967.2(R2=0.999 9),平均加样回收率分别为101.48%(n=6),RSD为0.26%;氟苯尼考纳米乳在50℃、75%相对湿度(RH)条件下稳定。结果表明:建立的含量检查方法专属性强、准确度高、精密度、重复性良好,适用于氟苯尼考纳米乳含量测定,制备的氟苯尼考纳米乳稳定性良好,强光对其含量略有影响。
In order to establish a HPLC method for determining the content and stability of florfenicol nanoemulsion,an Iner Sustain C18column( 4. 6 mm×250 mm,5 μm) was applied at 25 ℃ during the detection,and acetonitrile-water-phosphate( 35: 65: 0. 02,V/V/V)with a flow rate of 1. 0 m L/min was used as the mobile phase. The detection wavelength was 224 nm,and the injection volume was 20 μL.The specificity,linearity,precision and recovery of the determination method were measured,and the stability of florfenicol nanoemulsion was studied based on an influencing factor test and an acceleration test. The results presented a linear relationship with the regression equation of A = 49 295 C + 2 967. 2( R2= 0. 999 9),which was well established in a range of 4. 12-67. 74 μg/m L; and an average recovery of101. 48%( n = 6) with RSD of 0. 26% was obtained. The florfenicol nanoemulsion was stable under the conditions of 50 ℃ and 75% RH.This method of determination showed strong specificity,high accuracy and precision with good repeatability,and was applicable to detection of the content of florfenicol nanoemulsion. Florfenicol nanoemulsion prepared in this study was of high stability,although strong light had a slight effect on the concentration of florfenicol nanoemulsion.
In order to establish a HPLC method for determining the content and stability of florfenicol nanoemulsion,an Iner Sustain C18column( 4. 6 mm×250 mm,5 μm) was applied at 25 ℃ during the detection,and acetonitrile-water-phosphate( 35: 65: 0. 02,V/V/V)with a flow rate of 1. 0 m L/min was used as the mobile phase. The detection wavelength was 224 nm,and the injection volume was 20 μL.The specificity,linearity,precision and recovery of the determination method were measured,and the stability of florfenicol nanoemulsion was studied based on an influencing factor test and an acceleration test. The results presented a linear relationship with the regression equation of A = 49 295 C + 2 967. 2( R2= 0. 999 9),which was well established in a range of 4. 12-67. 74 μg/m L; and an average recovery of101. 48%( n = 6) with RSD of 0. 26% was obtained. The florfenicol nanoemulsion was stable under the conditions of 50 ℃ and 75% RH.This method of determination showed strong specificity,high accuracy and precision with good repeatability,and was applicable to detection of the content of florfenicol nanoemulsion. Florfenicol nanoemulsion prepared in this study was of high stability,although strong light had a slight effect on the concentration of florfenicol nanoemulsion.
引文
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[13]曾勇,董文婷,龚波,等.超高效液相色谱-串联质谱法快速测定禽蛋中氯霉素、氟苯尼考及其代谢物残留[J].中国兽药杂志,2017,51(12):42-46.
[14]魏海涛,唐兴刚,于文刚,等.高效液相色谱法测定氟苯尼考悬浮乳剂含量[J].中国兽药杂志,2009,43(5):21-23.
[2]Shin S J,Kang S G,Nabin R,et al.Evaluation of the antimicrobial activity of florfenicol against bacteria isolated from bovine and porcine respiratory disease[J].Vet Microbiol,2005,106(1/2)73-77.
[3]Park B K,Lim J H,Kim M S,et al.Pharmacokinetics of florfenicol and its metabolites,florfenicol amine,in dogs[J].Res Vet Sci,2008,84(1):85-89.
[4]杨昆,刘晗,罗璋,等.氟苯尼考新制剂[J].动物医学进展,2011,32(8):108-111.
[5]Al-Shahrani S,Naidoo V.Florfenicol induces early embryonic death in eggs collected from treated hens[J].BMC Vet Res,2015,11:213-220.
[6]Du X,Xia C,Shen J,et al.Characterization of florfenicol resistance among calf pathogenic Escherichia coli[J].FEMS Microbiol Lett,2004,236(2):183-189.
[7]Shen J,Wu X,Hu D et al.Pharmacokinetics of florfenicol in healthy and Escherichia coli-infected broiler chickens[J].Res Vet Sci,2002,73(2):137-140.
[8]周飞飞,李志林,苏荣国,等.氟苯尼考浸泡栉孔扇贝后各组织中含量的测定[J].中国海洋大学学报(自然科学版),2018(4):48-56.
[9]Atef M,El-Gendi A Y,Amer A M,et al.Effect of three anthelmentics on disposition kinetics of florfenicol in goats[J].Food Chem Toxicol,2010,48(12):3340-3344.
[10]Sun Y X,Zhao H Y,Shan Q,et al.Pharmacokinetics of florfenicol in crucian carp(Carassiusauratus cuvieri)after a single instramuscular or oral administration[J].J Vet Pharmacol Ther,2011,34(5):460-463.
[11]Lim J H,Kim M S,Hwang Y H,et al.Plasma and tissue depletion of florfenicol in olive flounder(Paralichthys olivaceus)after oral administration[J].Aquaculture,2010,307(1):71-74.
[12]Shen J S,Xia X,Jiang H Y,et al.Determination of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine in poultry and porcine muscle and liver by gas chromatography-negative chemical ionization mass spectrometry[J].J Chromatogra B,2009,877(14):1523-1529.
[13]曾勇,董文婷,龚波,等.超高效液相色谱-串联质谱法快速测定禽蛋中氯霉素、氟苯尼考及其代谢物残留[J].中国兽药杂志,2017,51(12):42-46.
[14]魏海涛,唐兴刚,于文刚,等.高效液相色谱法测定氟苯尼考悬浮乳剂含量[J].中国兽药杂志,2009,43(5):21-23.