HPLC-MS法同时测定健脾益肾丸中8个成分的含量

目的：建立高效液相色谱-串联质谱法同时测定健脾益肾丸中8个主要成分（丹参素钠、丹酚酸B、松果菊苷、毛蕊异黄酮葡萄糖苷、毛蕊花糖苷、甘草苷、黄芪甲苷、芒柄花素）的含量。方法：采用ZORBAX C₁₈色谱柱（250 mm×4.6 mm，5 μm），以乙腈（A）-10 mmol·L^-1乙酸铵溶液（B）为流动相，梯度洗脱（0~1.2 min，5% A；1.2~2 min，5% A→20% A；2~4 min，20% A→40% A；4~8 min，40% A；8~10 min，40% A→95% A；10~17 min，95% A；17~25 min，5% A），流速为0.8 mL·min^-1，进样量为5 μL；采用电喷雾离子源（ESI），SIM模式，正负离子同时检测方式。结果：丹参素钠、丹酚酸B、松果菊苷、毛蕊异黄酮葡萄糖苷、毛蕊花糖苷、甘草苷、黄芪甲苷和芒柄花素检测的线性范围分别为0.04~2.5 μg·mL^-1（r=0.999 7）、0.035~1.0 μg·mL^-1（r=0.999 5）、0.15~20 μg·mL^-1（r=0.999 2）、0.04~20 μg·mL^-1（r=0.999 8）、0.004~2.0 μg·mL^-1（r=0.999 9）、0.06~2.0 μg·mL^-1（r=0.999 4）、0.035~2.5 μg·mL^-1（r=0.999 8）和0.006~0.2（r=0.999 5）μg·mL^-1；精密度、稳定性、重复性试验的RSD小于3%；平均回收率（n=6）均在97.0%~102%范围内，RSD均小于3%。3批样品中丹参素钠、丹酚酸B、松果菊苷、毛蕊异黄酮葡萄糖苷、毛蕊花糖苷、甘草苷、黄芪甲苷和芒柄花素含量测定结果分别为0.490~0.500、1.792~1.812、0.505~0.511、0.680~0.684、0.609~0.615、0.100~0.110、0.056~0.057和0.020~0.022 mg·g^-1。结论：所建立可作为健脾益肾丸的质量控制提供依据。

Objective: To establish an HPLC-MS method for the determination of eight components in Jianpi Yishen pills(sodium danshensu,salvianolic acid B,echinacoside,calycosin 7-O-glucoside,acteoside,liquiritin, astragaloside Ⅳ).Methods: Chromatographic separation was carried out at 30℃ on a C₁₈ column(250 mm×4.6 mm,5 μm).The mobile phase consisted of acetonitrile(A)and 10 mmol·L^-1 ammonium acetate(B)using a gradient elution of 0-1.2 min,5% A;1.2-2 min,5% A → 20% A;2-4 min,20% A → 40% A;4-8 min,40% A;8-10 min,40% A → 95% A;10-17 min,95% A and 17-25 min,5% A with flow rate 0.8 mL·min^-1 The MS analysis worked in ESI positive and negative modes using selected ion monitoring mode.Results: The linear ranges were 0.04-2.5 μg·mL^-1(r=0.999 7)for sodium danshensu,0.035^-1.0 μg·mL^-1(r=0.999 5)for salvianolic acid B, 0.15-20 μg·mL^-1(r=0.999 2)for echinacoside, 0.04-20 μg·mL^-1(r=0.999 8)for calycosin 7-O-glucoside,0.004-2.0 μg·mL^-1(r=0.999 9)for acteoside,0.06-2.0 μg·mL^-1(r=0.999 4)for liquiritin, 0.035-2.5 μg·mL^-1(r=0.999 8)for astragaloside Ⅳ, 0.006-0.2 μg·mL^-1(r=0.999 5)for formononetin, and the average recovery was between 97.0% to 102%.The contents of sodium danshensu, salvianolic acid B, echinacoside,calycosin 7-O-glucoside, acteoside,liquiritin,astragaloside Ⅳ,formononetin were 0.490-0.500 mg·g^-1, 1.792-1.812 mg·g^-1, 0.505-0.511 mg·g^-1, 0.680-0.684 mg·g^-1, 0.609-0.615 mg·g^-1,0.100-0.110 mg·g^-1, 0.056-0.057 mg·g^-1 and 0.020-0.022 mg·g^-1, respectively.Conclusion: The method can be used for the quality control of Jianpi Yishen pills.

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