目的 考察适用于测定白脉复方脂质体（BM-Lip）包封率的方法。方法 以前期已成功制备的、同时包载了挥发油、姜黄素、甘草酸和甘草苷的BM-Lip为研究对象，以白脉复方中挥发油、甘草苷、甘草酸和姜黄素含量为评价指标，对超滤离心法、葡聚糖凝胶柱法、鱼精蛋白凝聚法、超速离心法4种分离方法在测定BM-Lip包封率时的适用性进行考察。结果 当采用超滤离心管（截留相对分子质量3 000）以13 000 r/min离心20 min时，游离药物中挥发油、甘草酸和姜黄素的回收率偏低；在以20%乙醇为洗脱剂、体积流量1.0 mL/min时，无论葡聚糖凝胶柱的径高比为1∶15还是1∶20，脂质体与游离挥发油的流出间隔时间均过短，且难以通过优化手段进行改善；此外，3种不同来源的鱼精蛋白以及鱼精蛋白的不同加入量（1～3倍体积量）对BM-Lip的凝聚度均未产生显著影响，而在4 ℃下17 000 r/min离心60 min时，虽然BM-Lip获得高凝聚度，但游离药物的回收率却均不符合测定要求；然而，在4 ℃下40 000 r/min高速离心60 min时，测得游离药物中挥发油、甘草苷、甘草酸和姜黄素的平均回收率在96.29%～101.95%，同时，空白脂质体与游离药物的物理混合物的平均加样回收率中，甘草苷、甘草酸和姜黄素在97.35%～106.10%，挥发油在80.22%～85.26%，符合测定要求。根据这些条件最终测得挥发油、甘草苷、甘草酸和姜黄素的包封率分别为（76.15±0.16）%、（10.15±1.41）%、（7.09±0.57）%和（84.08±1.03）%。结论 采用超速离心法时各成分的回收率测定结果良好，表明该分离方法可用于准确测定BM-Lip的包封率；为后续BM-Lip制剂过程的系统优化提供了重要基础，也为同时测定含有不同极性组分的中药复方脂质体包封率提供了有价值的参考。

Objective To investigate the method applicable to the determination of the encapsulation rate of Baimai Compound liposomes (BM-Lip). Methods The BM-Lip successfully prepared in the previous stage and simultaneously encapsulated with volatile oil, curcumin, glycyrrhizic acid and glycyrrhizin was used as the object of study, and the content of volatile oil, glycyrrhizic acid, glycyrrhizic acid and curcumin in the Baimai Compound was used as the index of evaluation, the applicability of four separation methods, including ultrafiltration centrifugation method, dextran gel elution, protamine aggregation method, and ultracentrifugation, for the determination of encapsulation rate of the BM-Lip was examined. Results The results showed that when a ultrafiltration centrifuge tube (3 000 relative molecular mass retained) was used to centrifuge the free drug at 13 000 r/min for 20 min, the recoveries of volatile oil, glycyrrhetinic acid, and curcumin were low; when 20% ethanol was used as the eluent at a volumetric flow rate of 1.0 mL/min, the elution time intervals between the liposomes and free volatile oils were too short, regardless of whether the diameter-to-height ratio of the dextran gel column was 1∶15 or 1∶20 and it was difficult to improve by optimization; moreover, the three different sources of protamine and the different addition amounts (1—3 times the volume) did not significantly affect the cohesion of BM-Lip, but when centrifuged at 17 000 r/min for 60 min at 4 ℃, although BM-Lip gained a high degree of cohesion, the recoveries of the free drug were not in compliance with the requirements of the assay; however, the recoveries of free drug were not in accordance with the requirements of the assay when centrifuged at 4 ℃ for 60 min at 40 000 r/min. However, when centrifuged at a high speed of 40 000 r/min for 60 min, the average recoveries of volatile oil, glycyrrhizin, glycyrrhetinic acid and curcumin in the free drug were measured between 96.29% and 101.95%, which met the requirements of determination. Meanwhile, the average spiked recoveries of the physical mixtures of blank liposomes and free drug ranged from 97.35% to 106.10% for glycyrrhizin, glycyrrhizic acid and curcumin, and from 80.22% to 85.26% for volatile oil. The encapsulation rates of volatile oil, glycyrrhizin, glycyrrhizic acid and curcumin were measured to be (76.15 ± 0.16)%, (10.15 ± 1.41)%, (7.09 ± 0.57)% and (84.08 ± 1.03)%, respectively, according to these conditions. Conclusion In conclusion, the good results of the recovery determination of each component by ultracentrifugation indicated that this separation method can be used to accurately determine the encapsulation rate of BM-Lip. This study not only provides an important basis for the systematic optimization of the subsequent process of BM-Lip, but also provides a valuable reference for the determination of the encapsulation rate of liposomes of traditional Chinese medicine compound prescriptions that contain different polar components at the same time.

R283.6

国家自然科学基金面上项目（82474201）；国家自然科学基金青年项目（82104545）；中央高校基本科研业务费专项资金资助（2023-JYB-JBQN-043）