中国药物警戒 ›› 2026, Vol. 23 ›› Issue (5): 533-538.
DOI: 10.19803/j.1672-8629.20250858

• 基础与临床研究 • 上一篇    下一篇

垂体后叶中活性成分测定方法的替代研究

张媛1, 郭龙静2△, 杨泽岸1, 贺庆1, 王宏宇1, 吴彦霖1#, 纳涛3,*   

  1. 1中国食品药品检定研究院化学药品检定所,北京 102629;
    2北京民海生物科技有限公司研发中心,北京 102609;
    3中国食品药品检定研究院生物制品检定所,北京 102629
  • 收稿日期:2025-12-01 发布日期:2026-05-20
  • 通讯作者: *纳涛,男,博士,副研究员,细胞检定。E-mail: natao@vip.163.com #为共同通信作者。
  • 作者简介:张媛,女,硕士,副研究员,药检药理。△为并列第一作者。
  • 基金资助:
    国家重点研发计划（2024YFA1107301）

Alternative Methodologies for Determination of Active Components in Posterrior Pituitary

ZHANG Yuan1, GUO Longjing2△, YANG Zean1, HE Qing1, WANG Hongyu1, WU Yanlin1#, NA Tao3,*   

  1. 1National Institutes for Food and Drug Control, Institute for Chemical Drug Control, Beijing 102629, China;
    2Research and Development Center, Beijing Minhai Biotechnology Co., Ltd., Beijing 102609, China;
    3National Institutes for Food and Drug Control, Institute for Biological Products Control, Beijing 102629, China
  • Received:2025-12-01 Published:2026-05-20

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摘要： 目的 建立高效液相色谱（High Performance Liquid Chromatography,HPLC）法测定垂体后叶中的主要活性成分赖氨酸升压素和缩宫素含量,并进行方法学验证,考察新建方法与《中华人民共和国药典》方法的相关性和等效性。方法 赖氨酸升压素采用C18柱（4.6 mm×25 cm,3.5 μm）,流动相A为0.1 mol·L-1磷酸氢二铵溶液（pH=6.0）,流动相B为乙腈和水（1∶1）混匀溶液,流速1.0 mL·min-1,检测波长220 nm,柱温40℃。缩宫素采用C18柱（4.6 mm×25 cm,5 μm）,流动相A为0.1 mol·L-1磷酸二氢钠溶液（pH=4.5）,流动相B为乙腈和水（1∶1）混匀溶液,流速1.0 mol·L-1,检测波长220 nm,柱温40℃。结果 所建的2种HPLC测定法中,赖氨酸升压素和缩宫素主峰与其相邻杂质峰之间的分离度均符合要求;在0.21~13.33 IU·mL-1内,升压素浓度与其色谱峰面积线性关系良好,r=0.999 9,在0.26~21 IU·mL-1内,缩宫素浓度与其色谱峰面积线性关系良好,r=0.999 9;平均回收率（n=9）分别为100.3%和99.8%;重复性精密度RSD分别为0.09%和0.44%;重现性精密度RSD分别为2.85%和1.9%。统计学结果表明,HPLC测定法与生物测定法具有良好的结果等效性和性能等效性。结论 所建方法操作简便、准确度高、重复性好且与生物测定法等效,可分别用于垂体后叶中赖氨酸升压素和缩宫素含量的测定。

关键词: 垂体后叶, 升压素, 缩宫素, 含量测定, 方法替代, 高效液相色谱, 等效性, 大鼠

Abstract: Objective To establish HPLC methods for the determination of active components—vasopressin and oxytocin—in posterior pituitary and to investigate the correlation and equivalence between the newly developed methods and those specified in the pharmacopoeia. Methods For vasopressin, a C18 column (4.6 mm×25 cm, 3.5 μm) was used. Mobile phase A consisted of 0.1 mol·L-1 diammonium hydrogen phosphate (pH=6.0) while mobile phase B was composed of a 1∶1 mixture of acetonitrile and water. The flow rate was 1.0 mL·min-1, detection wavelength 220 nm, and column temperature was 40°C. For oxytocin, a C18 column (4.6 mm×25 cm, 5 μm) was adopted. Mobile phase A was 0.1 mol·L-1 sodium dihydrogen phosphate solution (pH=4.5) while mobile phase B was a 1∶1 mixture of acetonitrile and water under the same conditions as vasopressin. Results Using either method, the resolution between the main peaks of vasopressin/oxytocin and adjacent impurity peaks met the required standard. A good linear relationship was observed between the concentration and chromatographic peak area: within the range of 0.21-13.33 IU·mL-1 for vasopressin (r=0.999 9), and 0.26-21 IU·mL-1 for oxytocin (r=0.999 9). The average recovery (n=9) was 100.3% and 99.8%, respectively. The RSD of repeatability precision was 0.09% and 0.44%, and that of reproducibility precision was 2.85% and 1.9%, respectively. Statistical analysis indicated good result equivalence and performance equivalence between the HPLC methods and the bioassay. Conclusion The established methods are user-friendly, accurate, reproducible, and equivalent to the bioassay, which can be used for determination of vasopressin and oxytocin contents in posterior pituitary.

Key words: Posterior Pituitary, Vasopressin, Oxytocin, Content Determination, Method Substitution, HPLC, Equivalence, Rats

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