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Volume 39 Issue 5
Sep.  2021
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Article Navigation >  Journal of Pharmaceutical Practice and Service  > 2021  >  39(5): 447-448, 453

CHENG Cheng, CHEN Xiangmei, MA Jian, ZHAO Chuan. Quantitative determination of quercetin in Honghuixiang injection by HPLC[J]. Journal of Pharmaceutical Practice and Service, 2021, 39(5): 447-448, 453. doi: 10.12206/j.issn.1006-0111.202103041
Citation: CHENG Cheng, CHEN Xiangmei, MA Jian, ZHAO Chuan. Quantitative determination of quercetin in Honghuixiang injection by HPLC[J]. Journal of Pharmaceutical Practice and Service, 2021, 39(5): 447-448, 453. doi: 10.12206/j.issn.1006-0111.202103041
shu

Quantitative determination of quercetin in Honghuixiang injection by HPLC

doi: 10.12206/j.issn.1006-0111.202103041

  • Department of Pharmacy , Medical Security Centre, Chinese PLA General Hospital, Beijing 100048, China

  • Received Date: 2021-03-19
  • Rev Recd Date: 2021-06-30
    • Available Online: 2021-09-28
  • Publish Date: 2021-09-25
  •   Objective  To establish quantitative methods to assay quercetin in Honghuixiang injection by HPLC.  Methods  Dikma C18 column(250 mm×4.6 mm, 5 μm) was used for the assay with acetonitrile −0.1% phosphoric acid (25∶75) as the mobile phase. Flow rate was 1.0 ml/min. The column temperature was 30 ℃. The detection wavelength was at 256 nm.  Results  Quercetin showed good linear relationship within the range of 0.2150–3.225 μg. The correlation coefficient was 0.999 6. The average recovery was 99.39% with RSD 0.82% (n=6). The repeatability was 1.194 mg/ml with RSD 0.40%.  Conclusion  The average quercetin content in three batches of Honghuixiang injection was 1.191 mg/ml. This method is simple, rapid and accurate. It can be used for the determination of quercetin in Honghuixiang injection.
  • [1] 国家药典委员会. 中华人民共和国药典(一部)[S]. 北京: 化学工业出版社, 2005.
    [2] 邱晓静, 苏文镜. HPLC法测定红茴香注射液中芦丁的含量[J]. 海峡药学, 2008, 20(3):70-71. doi:  10.3969/j.issn.1006-3765.2008.03.031
    [3] 王姿媛, 胡惠兰. HPLC法测定哥王药材中的槲皮苷含量[J]. 中国实验方剂学杂志, 2013, 19(7):127-129.
    [4] 常明泉, 潘涛. HPLC法测定氯定庆溶液中苯甲酸钠的含量[J]. 海峡药学, 2017, 29(3):60-62. doi:  10.3969/j.issn.1006-3765.2017.03.021
    [5] 董丹华, 刘玉军. HPLC法测定圣愈汤冻干粉中4个指标成分的含量[J]. 中国药房, 2020, 31(5):576-580. doi:  10.6039/j.issn.1001-0408.2020.05.14
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Quantitative determination of quercetin in Honghuixiang injection by HPLC

doi: 10.12206/j.issn.1006-0111.202103041
  • Department of Pharmacy , Medical Security Centre, Chinese PLA General Hospital, Beijing 100048, China
  • Received Date: 2021-03-19
  • Rev Recd Date: 2021-06-30
  • Available Online: 2021-09-28
  • Publish Date: 2021-09-25

Abstract:   Objective  To establish quantitative methods to assay quercetin in Honghuixiang injection by HPLC.  Methods  Dikma C18 column(250 mm×4.6 mm, 5 μm) was used for the assay with acetonitrile −0.1% phosphoric acid (25∶75) as the mobile phase. Flow rate was 1.0 ml/min. The column temperature was 30 ℃. The detection wavelength was at 256 nm.  Results  Quercetin showed good linear relationship within the range of 0.2150–3.225 μg. The correlation coefficient was 0.999 6. The average recovery was 99.39% with RSD 0.82% (n=6). The repeatability was 1.194 mg/ml with RSD 0.40%.  Conclusion  The average quercetin content in three batches of Honghuixiang injection was 1.191 mg/ml. This method is simple, rapid and accurate. It can be used for the determination of quercetin in Honghuixiang injection.

CHENG Cheng, CHEN Xiangmei, MA Jian, ZHAO Chuan. Quantitative determination of quercetin in Honghuixiang injection by HPLC[J]. Journal of Pharmaceutical Practice and Service, 2021, 39(5): 447-448, 453. doi: 10.12206/j.issn.1006-0111.202103041
Citation: CHENG Cheng, CHEN Xiangmei, MA Jian, ZHAO Chuan. Quantitative determination of quercetin in Honghuixiang injection by HPLC[J]. Journal of Pharmaceutical Practice and Service, 2021, 39(5): 447-448, 453. doi: 10.12206/j.issn.1006-0111.202103041
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  • 红茴香注射液是由红茴香根皮经提取制成的灭菌水溶液，具有消肿散瘀，活血止痛作用。临床用于腰肌劳损，关节或肌肉韧带伤痛等。总黄酮是红茴香注射液的主要成分。原标准以芦丁为对照品，采用紫外分光光度法测定总黄酮的量，专属性不强。故笔者参考《中国药典》[1] ，以槲皮苷为测定对象，采用HPLC法测定红茴香注射液中槲皮苷的含量[2-5]获得满意结果, 现报告如下。

    • 1.   仪器和试药

      1.1.   仪器

    • Agilent 1100系列型高效液相色谱仪；HP-Chemstation色谱工作站。

      • 1.2.   试药

    • 乙腈为色谱纯，其他试剂均为分析纯。槲皮苷对照品由中国药品生物制品检定所提供 (批号：111538-200301，含量测定用), 红茴香注射液由浙江泰康药业有限公司提供(批号：20040730)。

    2.   方法与结果

      2.1.   色谱条件

    • 色谱柱为Dikma C18(250 mm×4.6 mm，5 μm)；流动相为乙腈-0.1%磷酸水溶液(25∶75)，流速1.0 ml/min，柱温 30 ºC，检测波长256 nm，进样量为10 μl。色谱柱的理论塔板数按槲皮苷峰计算大于3 000。

      • 2.2.   溶液制备

      2.2.1.   对照品溶液

    • 精密称取槲皮苷对照品(120 ℃减压干燥至恒重)适量, 加甲醇制成每1 ml含0.1 mg 的溶液, 摇匀, 即得。

      • 2.2.2.   供试品溶液

    • 精密吸取本品3 ml，置25 ml 量瓶中，加甲醇至刻度，摇匀，用微孔滤膜(0.45 μm)滤过，取滤液作为供试品溶液。

      • 2.2.3.   阴性对照溶液

    • 取辅料作为缺样空白样品，按“2.2.2”项下方法制备。

      • 2.3.   专属性试验

    • 分别取阴性对照溶液，对照品溶液及供试品溶液适量，滤过取续滤液 10 μl 注入 HPLC 仪，分别记录色谱图。实验结果表明，该方法专属性良好，阴性对照无干扰，结果见图1。

      • 2.4.   线性关系考察

    • 精密称取经减压干燥24 h的槲皮苷对照品适量，加甲醇配成每l ml含槲皮苷0.215 0 mg的对照品溶液。分别精密吸取对照品溶液，注入高效液相色谱仪，按上述色谱条件进行测定，记录色谱图，以槲皮苷峰面积的积分值为Y，进样量为X(μg)，回归得线性方程Y=1 515.9X+57.088 2，r=0.999 8，表明在0.215～3.225 μg范围内呈良好线性关系。

      • 2.5.   重复性试验

    • 取同一批供试品6份（批号：20040730），分别按供试品溶液制备法制备，按“2.1”项色谱条件测定。结果表明含量值分别为：1.187、1.189、1.197、1.196、1.196、1.198 mg/ml，RSD为0.40%（n=6），表明此方法重复性较好。

      • 2.6.   精密度试验

    • 精密吸取同一供试品溶液（批号：20040730）10 μl，重复进样6次, 按“2.1”项下色谱条件测定。峰面积分别为2 214.51、2 217.06、2 232.75、2 236.86、2 250.09、2 190.69，RSD为0.86%（n=6），结果表明仪器精密度良好。

      • 2.7.   稳定性试验

    • 精密吸取同一批号供试品溶液（批号：20040730）10 μl，分别于0、0.5、1、2、8、12、24 h测定峰面积值，结果为2 220.96、2 223.36、2 230.08、2 235.78、2 246.88、2 200.44、2 227.17，平均峰面积为2 226.38，RSD为0.64%（n=6），结果表明供试品溶液在24 h内稳定。

      • 2.8.   加样回收率试验

    • 取已测定含量的样品6份，每份3 ml，分别加入槲皮苷对照品溶液10 ml（浓度为0.225 4 mg/ml），按供试品溶液的制备方法操作，进行回收率测定，结果分别为100.49%、99.33%、98.14%、99.82%、98.89%、99.65%，平均加样回收率为99.39%，RSD为0.82%（n=6）。

      • 2.9.   样品含量测定

    • 分别取3批不同批号的红茴香注射液样品，按“2.2”项下制备供试品溶液，样品测定结果见表1。

      批号槲皮苷平均含量
      200407301.1941.191
      200408241.186
      200412221.192
    3.   讨论

      3.1.   测定波长的选择

    • 取对照品溶液依法操作，在200～400 nm的波长范围内扫描，结果在256.6 nm处有最大吸收，因此选择256 nm作为测定波长(图2)。

      • 3.2.   溶剂的选择

    • 槲皮苷易溶于甲醇、乙醇, 用甲醇作溶剂的峰形比用乙醇作溶剂的峰形好, 因此选择用甲醇作为溶剂。

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