反相高效液相色谱法测定度鲁特韦钠的平衡溶解度

目的：建立反相高效液相色谱法测定度鲁特韦钠在不同介质中的平衡溶解度，为其剂型设计及生物药剂学分类提供依据。方法：用反相高效液相色谱法测定度鲁特韦钠的含量。色谱柱为C₁₈柱（4.6 mm×250 mm，5 μm）；流动相A为0.042 5 mol·L^-1磷酸盐缓冲液500 mL，用2 mol·L^-1氢氧化钠溶液调节pH至6.8，加入甲醇500 mL混匀，流动相B为1%磷酸乙腈溶液，A-B（85∶15）；流速为1.0 mL·min^-1；检测波长为260 nm；柱温为30℃。采用摇瓶法考察度鲁特韦钠在不同pH缓冲液中的平衡溶解度。结果：37℃时，度鲁特韦钠在pH 1.2、pH 4.5、pH 6.8缓冲液中的平衡溶解度分别为0.071 3、0.067 2、0.073 0 mg·mL^-1。结论：度鲁特韦钠几乎不溶于缓冲液，属于低溶解度药物，不可申请生物等效性试验豁免。 

Objective:To establish a RP-HPLC method to determine the equilibrium solubility of dolutegravir sodium in different mediums,so as to provide a basis for its formulation design and biopharmaceutical classification. Methods:The equilibrium solubility of dolutegravir sodium in different pH buffer solutions was determined by the RP-HPLC method combined with shaking flask. C₁₈ column(4.6 mm×250 mm,5 μm)was adopted. The mobile phase A was 0.042 5 mol·L^-1 phosphate buffer 500 mL,adjusted pH 6.8 with 2 mol·L^-1 sodium hydroxide solution,added methanol 500 mL and mixed. The mobile phase B was 1% phosphate acetonitrile solution. A-B(85:15)at the flow rate of 1.0 mL·min^-1. The detection wavelength was 260 nm,and the column temperature was 30℃. Results:Under the temperature of 37℃,the equilibrium solubilities of dolutegravir sodium in the pH 1.2,pH 4.5 and pH 6.8 buffer solutions were 0.071 3,0.067 2,0.073 0 mg·mL^-1,respectively. Conclusion:Dolutegravir sodium is almost insoluble in buffer solutions. It is a low solubility drug,and can not be authorized for a biowaiver of bioequivalence. 

[1] 闻家辰,赵临襄.Dolutegravir[J].中国药物化学杂志,2014,24(1):83 WEN JC,ZHAO LX.Dolutegravir[J].Chin J Med Chem,2014,24(1):83
[2] 杨臻峥.抗艾滋病药Dolutegravir[J].药学进展,2013,37(2):92 YANG ZZ.An anti-AIDS drug Dolutegravir[J].Prog Pharm Sci,2013,37(2):92
[3] 李艳玲,刘红淼,王彩霞,等.抗艾滋病新药-度鲁特韦[J].医药导报,2015,34(8):1064 LI YL,LIU HM,WANG CX,et al.A new anti-AIDS drug-dolutegravir[J].Her Med,2015,34(8):1064
[4] 赵长阔,王先恒,曹颖,等.抗艾滋病药物度鲁特韦的全球专利技术布局分析[J].中国新药杂志,2017,26(17):2003 ZHAO CK,WANG XH,CAO Y,et al.Research on global patent technology profile of dolutegravir:an anti-AIDS drug[J].Chin J New Drugs,2017,26(17):2003
[5] Martindale:The Complete Drug Reference.39th editon[S].2017:995
[6] 穆顺达,郑国华,宋成武,等.白杨素平衡溶解度、油水分配系数及解离常数的测定[J].药物分析杂志,2018,38(6):997 MU SD,ZHENG GH,SONG CW,et al.Measurement of equilibrium solubility,oil-water partition coefficients and dissociation constant of chrysin[J].Chin J Pharm Anal,2018,38(6):997
[7] 曾佳,黄婷,李芳,等.RP-HPLC测定左炔诺孕酮与孕二烯酮表观油水分配系数及平衡溶解度[J].中国新药杂志,2014,23(14):1703 ZENG J,HUANG T,LI F,et al.Determination of apparent oil-water partition coefficient and equilibrium solubility of levonorgestrel and gestodene by RP-HPLC[J].Chin J New Drugs,2014,23(14):1703
[8] DEZANI AB,PEREIRA TM,CAFFARO AM,et al.Equilibrium solubility versus intrinsic dissolution:characterization of lamivudine,stavudine and zidovudine for BCS classification[J].Braz J Pharm Sci,2013,49(4):853
[9] 中华人民共和国药典2015年版.二部[S].2015:凡例ⅩⅢ ChP 2015.Vol Ⅱ[S].2015:General Notice ⅩⅢ
[10] 金方方,尹婕,南楠.化学口服固体制剂仿制药质量和疗效一致性评价研究思考[J].药物分析杂志,2018,38(4):575 JIN FF,YIN J,NAN N.Study on quality and efficacy consistency evaluation of chemical oral solid generic drugs[J].Chin J Pharm Anal,2018,38(4):575
[11] Food and Drug Administration Center for Drug Evaluation and Research.Guidance for industry:"Waiver of in invo bioavailability and bioequivalence studies for immediate-release solid oral dosage forms based on a Biopharmaceutics Classification System"[EB/OL].Silver Spring.(2017-12-22)[2018-11-05].http://www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/default.htm
[12] 世界卫生组织.世界卫生组织药品标准专家委员会第40次技术报告[M].金少鸿,宁保明,译.北京:中国医药科技出版社,2009:333 WHO.WHO Expert Committee on Specifications for Pharmaceutical Preparations Technical Report Series Fortieth Report No.40[M].Translated by JIN SH,NING BM.Beijing:China Medical Science and Technology Press,2009:333
[13] European Medicines Agency.Note for guidance on the investigation of bioavailability and bioequivalence[EB/OL].London.(2001-07-26)[2018-11-05].https://www.ema.europa.eu/documents/scientific-guideline/note-guidance-investigation-bioavailability-bioequivalence_en.pdf
[14] 国家食品药品监督管理总局.人体生物等效性试验豁免指导原则(征求意见稿)[EB/OL].北京.(2016-04-08) [2018-11-05].http://samr.cfda.gov.cn/WS01/CL0778/149640.html China Food and Drug Administration.Human Bioequivalence Test Waiver Guidelines(Draft)[EB/OL].Beijing.(2016-04-08)[2018-11-05].http://samr.cfda.gov.cn/WS01/CL0778/149640.html
[15] AMIDON GL,LENNEMAS H,SHAH VP,et al.A theoretical basis for a biopharmaceutic drug classification:the correlation of in vitro drug product dissolution and in vivo bioavailability[J].Pharm Res,1995,12(3):413