目的 在遵循人用药品注册技术要求国际协调会(International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use,ICH)制剂稳定性指导原则的指导下,在采用超高效液相色谱-飞行时间质谱(UPLC-Q/TOF-MS)联用技术对中药传统水蜜丸制剂二至丸进行多成分定性定量研究的基础上,进行二至丸长期稳定性和加速稳定性研究,并考察了制剂影响因素对制剂稳定性的影响。方法 采用Acquity UPLC BEH C₁₈色谱柱(100 mm×2.1 mm,1.7 μm),流动相为0.1%甲酸水溶液-乙腈,梯度洗脱,体积流量0.2 mL/min,采用电喷雾电离源(ESI),正、负离子模式扫描。在遵循ICH制剂稳定性指导原则的指导下,进行了18个月的长期稳定性试验［温度(25±2)℃,相对湿度(RH)(60±5)%］,加速稳定性试验［温度(40±2)℃,RH(75±5)%］和制剂影响因素试验(包装材料类型和密封条件)。结果 采用以上方法共鉴定出二至丸中20个化学成分(红景天苷、蟛蜞菊内酯、10-羟基木犀榄苷二甲酯、木犀榄苷-11甲酯、马钱苷酸、松果菊苷、女贞苦苷、橄榄苦苷酸、毛蕊花苷/异毛蕊花苷、女贞苷、特女贞苷、ligustroflavone、特女贞苷异构体、阿富汗丁香苷F、异女贞苷、女贞酸、橄榄苦苷、女贞苷-G13、青藤碱、3-O-顺式-对-香豆酰委陵菜酸),并对其进行定量及归一化半定量研究。在长期稳定性研究中,20个成分均保持稳定;然而在加速稳定性试验条件下,11种成分(10-羟基木犀榄苷二甲酯、松果菊苷、女贞苦苷、橄榄苦苷酸、毛蕊花苷、女贞苷、特女贞苷、ligustroflavone、女贞酸、女贞苷-G13、青藤碱)的量明显下降。制剂影响因素试验结果显示采用钠钙玻璃包装材料替代现有市售聚酯包装,在密封或敞口条件下,制剂中各成分稳定性均显著提高。在采用相同包材时密封条件对制剂稳定性也存在显著性影响。结论 通过较为系统的中药传统口服制剂类型水蜜丸的制剂稳定性研究,为中药制剂质量控制水平和临床用药安全提供技术支持。

Objective According to the ICH guideline, the long-term stability and accelerated stability testing for Er-Zhi-Wan (water honey pills) has been carried out on the basis of the qualitative and quantitative analysis on multiple components in Er-Zhi-Wan by using UPLC-Q-TOF/MS method. In addition, the influence factors of the preparation (packing material and sealing process) were investigated by UPLC method. Methods The analysis was performed on an Acquity UPLC BEH C₁₈ column (100 mm × 2.1 mm, 1.7 μm) with the mixture of acetonitrile-water-formic acid as mobile phase, the flow rate was 0.2 mL/min and MS scanning mode was positive and negative. According to the ICH guideline, the 18 months long-term stability [(25 ± 2) ℃, relative humidity (RH) of (60 ± 5)%], accelerated stability testing [(40 ± 2) ℃, RH of (75 ± 5)%], and influence factors of preparation (packing material and sealing process) for Er-Zhi-Wan (water honey pills) have been carried out by the UPLC method. Results A total of 20 chemical compounds (salidroside, wedelolactone, 10-hydroxyoleoside dimethyl ester, oleoside-11 methyl ester, loganic acid, echinacoside, nuezhengslaside, oleuropein acid, verbascoside/isoverbascoside, nuezhenoside, specnuezhenide, ligustroflavone, isomer of specnuezhenide, safghanoside F, isonuezhenide, nuezhenidic acid, oleuropein, nuezhenoside-G13, oleonuezhenide, and 3-O-cis-p-coumaroyltormentic acid) were identified in the qualitative study. The content of these compounds was measured by the established quantitative and semi-quantitative research methods. In long-term stability testing, the 20 compounds were all remained stable. However, in the accelerated testing, the content of 11 chemical compounds (10-hydroxyoleoside dimethyl eser, echinacoside, nuezhengslaside, oleuropein acid, verbascoside, nuezhenide, specnuezhenide, ligustroflavone, nuezhenidic acid, nuezhenoside-G13, and oleonuezhenide) decreased obviously. Taking soda-lime glass as the packaging material, the stability of Er-Zhi-Wan (water honey pills) was improved than the commercial package under the same sealing conditions. The airtightness of the packaging materials played a significant part on the pill's stability under different sealing conditions. Conclusion The study offers the scientific and technical support for the quality control and the clinical safety of the preparation in Chinese materia medica.

国家自然科学基金资助项目(81473543);重大新药创制项目(2014BAI05B01)