目的 建立熱痹痛合劑的HPLC指紋圖譜和熱痹痛合劑中芍藥苷、柚皮苷、黃芩苷、川續(xù)斷皂苷VI的HPLC測(cè)定方法。方法 指紋圖譜分析采用Agilent 1260 C18色譜柱(250 mm×4.6 mm, 5μm),以乙腈-0.2%磷酸溶液為流動(dòng)相,梯度洗脫;檢測(cè)波長(zhǎng)為212 nm;體積流量為1 mL/min;柱溫30℃;進(jìn)樣量10μL。對(duì)指紋圖譜進(jìn)行相似度評(píng)價(jià),并運(yùn)用聚類(lèi)分析、主成分分析、正交偏最小二乘法-判別分析分析熱痹痛合劑樣品圖譜。采用HPLC法測(cè)定芍藥苷、柚皮苷、黃芩苷、川續(xù)斷皂苷VI,采用Agilent 1260 C18色譜柱(250 mm×4.6 mm, 5μm),流動(dòng)相為乙腈-0.2%磷酸溶液,梯度洗脫;檢測(cè)波長(zhǎng)230 nm(芍藥苷)、280 nm(柚皮苷、黃芩苷)、260 nm(川續(xù)斷皂苷VI);體積流量1 mL/min;柱溫30℃;進(jìn)樣量為10μL。結(jié)果 10批熱痹痛合劑樣品的指紋圖譜中有26個(gè)共有峰,相似度均大于0.99,化學(xué)模式識(shí)別將10批樣品分為2大類(lèi), OPLS-DA分析篩選出4個(gè)對(duì)制劑質(zhì)量差異影響較大的化合物。含量測(cè)定中芍藥苷、柚皮苷、黃芩苷、川續(xù)斷皂苷VI分別在27.238~272.384、38.207~382.066、77.864~778.642、51.015~510.015μg/mL線性良好,平均回收率分別為95.34%、103.51%、103.51%、102.68%, RSD值分別0.65%、1.12%、0.86%、2.41%。結(jié)論 所建立的方法準(zhǔn)確、合理,重復(fù)性好,可用于熱痹痛合劑的質(zhì)量控制和評(píng)價(jià)。

Objective To establish the HPLC fingerprint of Rebitong Mixture and to simultaneously determine the contents of paeoniflorin, naringin, baicalin, and asperosaponin Ⅵ in Rebitong Mixture by HPLC method.Methods The fingerprint analysis was adopted on Agilent 1260 C₁₈ (250 mm×4.6mm, 5 μm) with the mobile phase consisted of acetonitrile - 0.2% phosphoric acid flowing at 1.0 mL/min in gradient elution manner, and the detection wavelength was set at 212 nm with column temperature 30 ℃. Similarity of fingerprints was evaluated, and cluster analysis (CA), principal component analysis (PCA) and orthogonal partial least squaresdiscriminant analysis (OPLS-DA) were used to analyze the sample patterns. The contents of paeoniflorin, naringin, baicalin, and asperosaponin Ⅵin Rebitong Mixture were determined by HPLC method on Agilent 1260 C₁₈ column (250 mm×4.6 mm, 5 μm). Acetonitrile - 0.2% phosphoric acid was used as the mobile phase by gradient elution. The detection wavelength were 230 nm for paeoniflorin, 280 nm for naringin and baicalin, and 260 nm for asperosaponin Ⅵ. The flow rate was 1 mL/min, the column temperature was 30 ℃, and the injection volume was 10 μL.Results There were 26 common peaks in the fingerprints of 10 batches of Rebitong Mixture samples, and the similarity was all above 0.99. The 10 batches of samples were divided into 2 categories with chemical pattern recognition, and four peaks that had a greater impact on the quality of preparations were selected by OPLS-DA analysis. The linearity of paeoniflorin, naringin, baicalin, and asperosaponin Ⅵ was 27.238 — 272.384, 38.207 — 382.066, 77.864 — 778.642, 51.015 — 510.015 μg/mL, and the average recoveries were 95.34%, 103.51%, 103.51%, and 102.68% with RSD values of 0.65%, 1.12%, 0.86%,and 2.41%, respectively.Conclusion The method is accurate, reasonable, and reproducible, and can be used for the quality control and evaluation of Rebitong Mixture.

R286.02

廣西中醫(yī)藥壯瑤醫(yī)藥院內(nèi)制劑孵化基地項(xiàng)目(桂中醫(yī)藥發(fā)[2021]7號(hào))