目的 建立三七总皂苷(Panax notoginseng saponins,PNS)传递体(transfersomes,TFSs)(PNS-TFSs)中5种皂苷类成分含量与包封率的测定方法,并探讨药物包封特性。方法 采用超高效液相色谱(UPLC)法测定PNS-TFSs制剂中三七皂苷R₁(NGR₁)、人参皂苷Rg₁(GRg₁)、人参皂苷Re(GRe)、人参皂苷Rb₁(GRb₁)与人参皂苷Rd(GRd)的含量,色谱柱为Hypersil Gold柱(100 mm×2.0 mm,1.9 μm),流动相为乙腈-水,梯度洗脱,检测波长为203 nm,柱温为28℃。以离心超滤法结合UPLC测定传递体包封率。结果 NGR₁、GRg₁、GRe、GRb₁与GRd对应的各色谱峰专属性与分离度良好(R ≥ 1.5),且分别在4.04~505.00、3.98~498.00、4.03~504.00、3.99~499.00、4.00~500.00 μg/mL呈良好的线性关系(r ≥ 0.999 7),精密度(RSD ≤ 2.40%)、准确度(97.23% ≤ 回收率 ≤ 104.50%)与供试品溶液稳定性(RSD ≤ 0.90%)均符合要求;测得PNS传递体中NGR₁、GRg₁、GRe、GRb₁与GRd质量浓度依次为98.14、380.80、41.68、317.50、75.61 μg/mL,包封率依次为75.48%、69.68%、69.51%、92.35%、95.97%。结论 UPLC法与离心超滤法可用于PNS传递体中多成分含量与包封率的测定,方法快速、准确、可靠。

Objective To develop methods for the determination of contents and encapsulation efficiencies of five saponins in Panax notoginseng saponins (PNS) transfersomes (PNS-TFSs) and probe the drug encapsulation features. Methods UPLC was adopted to determine the contents of notoginsenoside R₁ (NGR₁), ginsenoside Rg₁ (GRg₁), ginsenoside Re (GRe), ginsenoside Rb₁(GRb₁) and ginsenoside Rd (GRd) in the preparation of PNS-TFSs. An Hypersil Gold column (100 mm×2.0 mm, 1.9 μm) was used to separate the analytes with acetonitrile-water mixture as the mobile phase in gradient elution mode, and the detection wavelength was set at 203 nm, the column temperature was 28℃. Encapsulation efficiencies were determined by centrifugal ultrafiltration method combined with UPLC. Results The specificity and resolution (R ≥ 1.5) of the peaks corresponding to each analyte met requirements of methodology. The calibration curves were linear (r ≥ 0.9997) and in the ranges of 4.04-505.00, 3.98-498.00, 4.03-504.00, 3.99-499.00, 4.00-500.00 μg/mL for NGR₁, GRg₁, GRe, GRb₁ and GRd respectively. RSD (≤ 2.4%) of repeated measurements with working solution of chemical reference substances (CRS) and recoveries (97.23%-104.50%) of the analytes from the blank transfersomes spiked with their CRS demonstrated respectively the precision and accuracy of the method. The test solutions were stable (RSD ≤ 0.90%) in 12 h. The contents of NGR₁, GRg₁, GRe, GRb₁ and GRd in the transfersomes were 98.14, 380.80, 41.68, 317.50, 75.61 μg/mL, and their encapsulation efficiencies were 75.48%, 69.68%, 69.51%, 92.35%, 95.97%, respectively. Conclusion UPLC is fast, accurate, precise and applicable to the determination of the contents of NGR₁, GRg₁, GRe, GRb₁ and GRd in the transfersomes and the centrifugal ultrafiltration method coupled with UPLC is applicable to the determination of their encapsulation efficiencies.

R283.6

国家自然科学基金资助项目（82174096）；校级科研基金（国家自然科学基金预研专项）资助项目（2019ZG37）