酒苁蓉HPLC特征图谱及5个苯乙醇苷类成分含量测定

目的：采用高效液相色谱法建立酒苁蓉的特征图谱，并同时测定5个苯乙醇苷类成分的含量。方法：以松果菊苷、肉苁蓉苷A、毛蕊花糖苷、异毛蕊花糖苷和2'-乙酰基毛蕊花糖苷为对照，采用Agilent EclipseXDB-C₁₈（4.6 mm×250 mm，5 μm）色谱柱，以甲醇为流动相A，0.1%甲酸为流动相B，梯度洗脱（0~15min，15% A → 21% A；15~20 min，21% A → 25% A；20~70 min，25% A → 43% A），流速1.0 mL·min^-1，检测波长为330 nm，柱温35℃。结果：建立酒苁蓉特征图谱，70 min内酒苁蓉的主要色谱峰能够达到完全分离，同时标定5个苯乙醇苷类成分，对15批饮片中的5个成分进行含量测定。松果菊苷、肉苁蓉苷A、毛蕊花糖苷、异毛蕊花糖和2'-乙酰基毛蕊花糖苷的质量浓度分别在10.05~1 005 μg·mL^-1（r=0.999 9，n=6）、2.02~202 μg·mL^-1（r=0.999 9，n=6）、2.04~204 μg·mL^-1（r=0.999 9，n=6）、2.01~201 μg·mL^-1（r=0.999 9，n=6）、2.02~202 μg·mL^-1（r=0.999 9，n=6）范围内线性关系良好，方法的平均回收率（n=6）分别为99.8%（RSD=0.30%）、99.5%（RSD=1.6%）、100.2%（RSD=0.44%）、99.5%（RSD=0.53%）、99.8%（RSD=0.36%）；15批酒苁蓉中上述5个成分含量分别为0.20%~7.38%、0.03%~0.20%、0.03%~0.72%、0.03%~0.75%、0.03%~0.07%。结论：该方法操作简单，重复性好，并能同时测定5个苯乙醇苷类成分含量，具备定性定量双重作用，可为酒苁蓉的质量标准研究提供更加全面的依据。

Objective: To establish an HPLC characteristic chromatogram and quantitative method for five components in Cistanches Herba. Methods: With echinacoside, cistanoside A, acteoside, iso-acteoside and 2'-acetyacteoside as reference substances,HPLC analysis was performed on an Agilent Eclipse XDB-C₁₈ column (4.6 mm×250 mm, 5 μm). The mobile phase was methanol-0.1% formic acid solution with gradient elution (0-15 min, 15%A → 21%A; 15-20 min, 21%A → 25%A; 20-70 min, 25%A → 43%A). The flow rate was 1 mL·min^-1, the detection wavelength was 330 nm and the column temperature was 35℃. Results: HPLC characteristic chromatogram of Cistanches Herba was established and five characteristic peaks were marked. Five characteristic peaks were simultaneously determined by HPLC within 70 min. The linear ranges of echinacoside, cistanoside A, acteoside, iso-acteoside and 2' -acetyacteoside were 10.05-1005 μg·mL^-1(r=0.999 9, n=6), 2.02-202 μg·mL^-1(r=0.999 9, n=6), 2.04-204 μg·mL^-1(r=0.999 9, n=6), 2.01-201 μg·mL^-1 (r=0.999 9, n=6) and 2.02-202 μg·mL^-1(r=0.999 9, n=6), respectively. The average recovery rate (n=6) were 99.8%(RSD=0.30%), 99.5%(RSD=1.6%), 100.2%(RSD=0.44%), 99.5%(RSD=0.53%) and 99.8%(RSD=0.36%), respectively. The contents of the five components in 15 batches of Cistanches Herba were 0.20%-7.39%, 0.03%-0.20%, 0.03%-0.72%, 0.03%-0.75% and 0.03%-0.07%, respectively. Conclusion: The proposed specific HPLC characteristic chromatogram and quantitation method of five components for Cistanches Herba is simple and repeatable, which offer reference for quality control of the drug.

[1] 中华人民共和国药典2015年版. 一部[S]. 2015:216 ChP 2015. Vol Ⅰ[S]. 2015:216
[2] 雷丽,宋志宏,屠鹏飞. 肉苁蓉属植物的化学成分研究进展[J]. 中草药,2003, 34(5):473 LEI L, SONG ZH, TU PF. Advances in research of chemical constituents in plants of Cistanche Hoffing. et Link.[J]. Chin Tradit Herb Drugs,2003, 34(5):473
[3] 曹振杰,赵文军,吴雪萍. 肉苁蓉中苯乙醇甙类化合物的LC-MS分析[J]. 中药材, 2004, 27(3):175 CAO ZJ, ZHAO WJ, WU XP. Determination of phenylethanoid glycosides from Cistanche deserticola in spring and autumn with LC-MS[J]. J Chin Med Mater, 2004, 27(3):175
[4] 宋志宏,雷丽,屠鹏飞. 肉苁蓉属植物的药理活性研究进展[J]. 中草药,2003,34(9):113 SONG ZH,LEI L,TU PF. Advances in research of pharmacological activity in plants of Cistanche Hoffing. et Link.[J]. Chin Tradit Herb Drugs,2003, 34(9):113
[5] 马志国. 沙苁蓉与肉苁蓉饮片HPLC指纹图谱鉴别研究[J]. 中国药学杂志, 2011, 46(12):899 MA ZG. Differentiation of processed Cistanche sinensis G. Beck and Herba Cistanches by HPLC[J]. Chin Pharm J,2011, 46(12):899
[6] 朱乃亮,徐荣,吴海峰,等. 荒漠肉苁蓉和管花肉苁蓉指纹图谱比较研究[J]. 中国药学杂志, 2016, 51(13):1116 ZHU NL,XU R, WU HF,et al. Fingerprint comparative analysis of Cistanche deserticola Y. C. Ma and Cistanche tubulosa(Schrenk) Wight[J]. Chin Pharm J,2016, 51(13):1116
[7] 李承花,刘婧慧,徐珍霞,等. RP-HPLC同时测定管花肉苁蓉中4种苯乙醇苷的含量[J]. 药物分析杂志, 2010, 30(6):1003 LI CH, LIU JH, XU ZX, et al. RP-HPLC simultaneous determination of four phenylethanoid glycosides in Cistanche tubulosa (Schrenk)[J]. Chin J Pharm Anal,2010, 30(6):1003
[8] 彭芳,徐荣,王夏,等. 不同等级整株肉苁蓉的药材品质和产地加工蒸制时间研究[J]. 中药材,2016, 39(2):302 PENG F,XU R,WANG X,et al. Effect of grade and steaming time on quality of cultivated Cistanche deserticola[J]. J Chin Med Mater, 2016, 39(2):302
[9] 马志国,谭咏欣. 酒蒸不同时间肉苁蓉中6种苯乙醇苷类成分的变化[J]. 中成药,2011, 33(11):1951 MA ZG,TAN YX. Contents changes of six phenylethanoid glycosides under steaming time spans with wine in desertliving Cistanche[J]. Chin Tradit Pat Med,2011, 33(11):1951
[10] 钱浩,喻芳君,耿宗成,等. 不同季节采收的肉苁蓉中4种苯乙醇苷的含量比较研究[J]. 药物分析杂志,2016, 36(11):1971 QIAN H,YU FJ,GENG ZC,et al. Comparative study of the content of four phenylethanoid glycosides in Cistanche deserticola Y. C. Ma from different seasons[J]. Chin J Pharm Anal,2016,36(11):1971