RP-HPLC测定骨筋丸胶囊中羟基红花黄色素A、落干酸和龙胆苦苷的含量

目的：建立HPLC法测定骨筋丸胶囊中羟基红花黄色素A、落干酸和龙胆苦苷含量。方法：测定羟基红花黄色素A，采用Eclipse XDB-C₁₈色谱柱（250 mm×4.6 mm，5 μm），以甲醇-水-磷酸（27：73：0.05）为流动相，流速1.0 mL·min^-1，检测波长403 nm，柱温为室温（20 ℃）；测定落干酸和龙胆苦苷，采用Eclipse XDB-C₁₈色谱柱（250 mm×4.6 mm，5 μm），以乙腈-水（10：90，醋酸调pH 2.5）为流动相，流速1.0 mL·min^-1，检测波长250 nm，柱温35 ℃。结果：羟基红花黄色素A、落干酸和龙胆苦苷的线性范围分别为0.014~0.288 μg（r=0.9995）、0.056~1.120 μg（r=0.9998）和0.114~2.284 μg（r=0.9997），平均加样回收率（n=9）分别为98.8%、99.8%和99.3%。结论：所建立的方法经方法学验证快速简便，重复性好，专属性强，可作为中药制剂骨筋丸胶囊的质量控制方法。

Objective: To develop HPLC methods for the content determination of hydroxysafflor yellow A,loganic acid and gentiopicroside in Gujinwan capsules. Methods: The separation was performed on Eclipse XDB-C₁₈ column (250 mm×4.6 mm,5 μm) at room temperature (20 ℃).The mobile phase was composed of methanol-water-phosphoric acid (27:73:0.05) for the determination of hydroxysafflor yellow A at a flow rate of 1.0 mL·min^-1,and the detection wavelength was at 403 nm.The separation was performed on Eclipse XDB-C₁₈column (250 mm×4.6 mm,5 μm) at 35 ℃.The mobile phase was composed of acetonitrile-water (10:90,adjusted pH to 2.5 with acetic acid) for the determination of loganic acid and gentiopicroside at a flow rate of 1.0 mL·min^-1,and the detection wavelength was at 250 nm. Results: The calibration curves were linear within the range of 0.014-0.288 μg (r=0.9995),0.056-1.120 μg (r=0.9998) and 0.114-2.284 μg (r=0.9997) for hydroxysafflor yellow A,loganic acid and gentiopicroside,respectively. The average recoveries were 98.8%,99.8% and 99.3% for hydroxysafflor yellow A,loganic acid and gentiopicroside,respectively. Conclusion: The method is applicable for the determination of hydroxysafflor yellow A,loganic acid and gentiopicroside in Gujinwan capsules by methodology validation.

[1] SDA(国家药品监督管理局).Department of Orthopaedics and Traumatology(骨伤科分册)[G]//The National Standard for Traditional Chinese Medicines Assembly(国家中成药标准汇编).2002:252
[2] ChP 2010.Vol Ⅰ(中国药典2010年版.一部)[S].2010:141, 253
[3] JIA Na(贾娜), ZHANG Ya-hui(张雅惠), WANG Bin(王斌), et al.Determination of loganic acid and gentiopicroside and study on HPLC specific chromatogram of Gentiana straminea Maxim.(麻花秦艽中落干酸和龙胆苦苷的含量测定及麻花秦艽药材HPLC特征图谱的研究)[J].Chin J Pharm Anal(药物分析杂志), 2009, 29(9):1357
[4] WANG Jian-jun(汪建君), LI Ling-ling(李玲玲), CHEN Hui-ling(陈惠玲), et al.HPLC determination of hydroxysaffler yellow A in Zhengtian pills and Zhengtian capsules(HPLC法测定正天丸和正天胶囊中羟基红花黄色素A的含量)[J].Drug Stand China(中国药品标准), 2013, 14(3):197
[5] WANG Xiao-ping(王小平), LIU Feng(刘峰), MA Cun-de(马存德), et al.Improvement on testing method of hydroxysafflor yellow A from Honghua(safflower)(红花中羟基红花黄色素A含量测定方法的改进)[J].J Shaanxi Coll Chin Med(陕西中医学院学报), 2010, 33(4):96
[6] SUN Jing(孙菁), CHEN Gui-chen(陈桂琛), LI Yu-lin(李玉林).Determination of four iridoid glycosides in Tibetan mendicine Gentiana straminea Maxim.by HPLC(藏药麻花艽中四种苦苷类化学成分的HPLC测定)[J].Chin J Anal Lab(分析实验室), 2006, 25(5):28
[7] CHEN Yue-rong(陈岳蓉), LIU Hai-qing(刘海青), SU Dong-min(苏东敏).HPLC simultaneous determination of loganic acid and gentiopicroside in Gucixiaotong capsules(HPLC法同时测定骨刺消痛胶囊中落干酸和龙胆苦苷的含量)[J].Chin J Pharm Anal(药物分析杂志), 2009, 29(7):1169
[8] YU Qing-fen(俞青芬).Compare with the active constituents in different districts of Gentiana macrophylla Pall.(青海不同地区秦艽、麻花艽中落干酸和龙胆苦苷的含量测定)[J].J Jiangxi Norm Univ(Nat Sci)(江西师范大学学报自然科学版), 2010, 34(2):174
[9] MA Xiao-qing(马晓庆), DENG Wei-feng(邓伟峰), YANG Dong-hui(杨东辉).HPLC fingerprinting and quantification of gentiopicroside and loganic acid in Gentianae Macrophyllae Radix crude drugs(秦艽生药中龙胆苦苷和落干酸的含量及其HPLC指纹图谱测定)[J].J Chin Pharm Sci(中国药学英文版), 2010, (19):243