多肽微球注射剂中甘露醇含量测定研究

目的： 建立高效液相色谱-示差折光法测定注射用醋酸亮丙瑞林和醋酸奥曲肽微球中甘露醇含量。方法： 采用Phenomenex Rezex RHM-Monosaccharide H⁺（8%）强阳离子交换色谱柱（300 mm×7.8 mm）；流动相为超纯水；流速0.5 mL·min^-1，检测器为示差检测器，内部温度45℃，柱温60℃，进样量20 μL。结果： 甘露醇进样量在1.01~50.44 μg范围内线性关系良好（r²=1.000），精密度（RSD=0.15%）、稳定性（RSD=0.38%）和重复性（RSD=1.7%）良好，平均回收率为100.4%（n=9，RSD=0.89%）。醋酸亮丙瑞林微球粉末的甘露醇含量范围为3.08%~17.41%，醋酸奥曲肽微球粉末的甘露醇含量范围为16.03%~16.95%。结论： 本文建立的方法适用于多肽微球注射剂中甘露醇的含量测定。

Objective: To establish an HPLC-RI method for determination of mannitol in leuprorelin acetate and octreotide acetate microspheres for injection. Methods: The Phenomenex Rezex RHM-Monosaccharide H⁺(8%)column (300 mm×7.8 mm) was used,the mobile phase was water at a flow rate of 0.5 mL·min^-1. The detector was refractive index detector with inner temperature set at 45℃. The column temperature was 60℃,and the injection volume was 20 μL. Results: The linear range of mannitol was 1.01-50.44 μg(r²=1.000). The precision (RSD=0.15%),stability (RSD=0.38%) and repeatability (RSD=1.7%) tests were good. The average recoveries were 100.4% (n=9,RSD=0.89%). The contents of mannitol of leuprorelin acetate microspheres were among 3.08%-17.41%. The contents of mannitol of octreotide acetate microspheres were among 16.03%-16.95%. Conclusions: The established method can be used for determination of mannitol in injectable peptide microspheres.

[1] 陆丽芳,潘俊,陆伟跃.多肽及蛋白类药物微球的研究进展[J].中国医药工业杂志,2004,35(2):113 LU LF,PAN J,LU WY.Development of research on peptide-protein microspheres[J].Chin J Pharm,2004,35(2):113
[2] SANDERS LM,MCRAE GI,VITALE KM,et al.Controlled delivery of an Lhrh analogue from biodegradable injectable microspheres[J].J Controlled Release,1985,2:187
[3] SHIRANGI M,HENNINK WE,SOMSEN GW,et al.Identification and assessment of octreotide acylation in polyester microspheres by LC-MS/MS[J].Pharm Res,2015,32(9):3044
[4] YANG YY,CHUNG TS,NG NP.Morphology,drug distribution,and in vitro release profiles of biodegradable polymeric microspheres containing protein fabricated by double-emulsion solvent extraction/evaporation method[J].Biomaterials,2001,22(3):231
[5] OKADA H.One- and three-month release injectable microspheres of the LH-RH superagonist leuprorelin acetate[J].Adv Drug Del Rev,1997,28(1):43
[6] 陈燕娜,栾瀚森,王浩.甘露醇对地塞米松PLGA植入剂体外释放行为的影响[J].中国医药工业杂志,2017,48(8):1148 CHEN YN,LUAN HS,WANG H.Investigation on effect of mannitol on in vitro release behavior of dexamethasone plga implants[J].Chin J Pharm,2017,48(8):1148
[7] 刘薇,肖新月,宋宗华,et al.HPLC-RID法替代滴定法测定百令胶囊中甘露醇含量的研究[J].药物分析杂志,2014,34(1):159 LIU W,XIAO XY,SONG ZH,et al.Research on determination of mannitol in Bailing capsules by HPLC-RID instead of titration[J].Chin J Pharm Anal,2014,34(1):159
[8] 杨昕,涂秩平,李高.HPLC-ELSD法测定不同来源人工蛹虫草子实体中甘露醇的含量[J].药物分析杂志,2009,29(10):1668. YANG X,TU YP,LI G.HPLC-ELSD determination of mannitol in cultured Cordyceps militaris L.Link.[J].Chin J Pharm Anal,2009,29(10):1668
[9] 郭宁子,辛中帅,孙悦,等.激光散射法测定注射用醋酸亮丙瑞林微球粒径及其分布[J].中国新药杂志,2017,26(8):886 GUO NZ,XIN ZS,SUN Y,et al.Determination of size and size distribution of leuprorelin acetate microspheres for injection using laser diffraction method[J].Chin J New Drugs,2017,26(8):886
[10] 郭宁子,辛中帅,杨化新.微球制剂质量控制研究进展[J].中国新药杂志,2015,24(18):2115 GUO NZ,XIN ZS,YANG HX.Research progress in quality control of microspheres[J].Chin J New Drugs,2015,24(18):2115
[11] KUMAR R,PALMIERI MJ.Points to consider when establishing drug product specifications for parenteral microspheres[J].Aaps J,2010,12(1):27
[12] 李勋,韦祎,马光辉,胡琳琳.缓释微球制剂的研究进展[J].北京化工大学学报,2017,44(6):1 LI X,WEI Y,MA GH,et al.Recent research and development prospects for sustained-release microspheres[J].Beijing Univ Chem Technol,2017,44(6):1