HPLC同时测定草珊瑚中9个有效成分的含量

目的：建立HPLC法同时测定草珊瑚中落新妇苷、新绿原酸、绿原酸、隐绿原酸、咖啡酸、槲皮素-3-O-β-D-葡萄糖醛酸苷、异嗪皮啶、山柰酚-3-O-β-D-葡萄糖醛酸苷和迷迭香酸的含量。方法：采用Diamonsil 十八烷基硅烷键合硅胶（C₁₈）色谱柱（250 mm×4.6 mm，5 μm），流动相为乙腈（A）-0.2%磷酸水溶液（B），梯度洗脱（0~20 min，2% A→15% A；20~23 min，15% A→12.5% A；23~42 min，12.5% A；42~70 min，12.5% A→30% A），流速1.0 mL·min^-1，检测波长为290 nm（落新妇苷）、344 nm（新绿原酸、绿原酸、隐绿原酸、咖啡酸、槲皮素-3-O-β-D-葡萄糖醛酸苷、异嗪皮啶、山柰酚-3-O-β-D-葡萄糖醛酸苷和迷迭香酸），柱温35 ℃。结果：落新妇苷、新绿原酸、绿原酸、隐绿原酸、咖啡酸、槲皮素-3-O-β-D-葡萄糖醛酸苷、异嗪皮啶、山柰酚-3-O-β-D-葡萄糖醛酸苷和迷迭香酸分别在0.041~2.050、0.036 0~1.780、0.046~2.282、0.047~2.330、0.040~2.010、0.021~1.060、0.047~2.335、0.024~1.220、0.047~2.368 μg范围内与峰面积呈现良好的线性关系（r=0.999 8~1.000），平均加样回收率（n=6）在98.4%~101.7%之间，RSD为1.1%~2.7%。17批草珊瑚样品中落新妇苷、新绿原酸、绿原酸、隐绿原酸、咖啡酸、槲皮素-3-O-β-D-葡萄糖醛酸苷、异嗪皮啶、山柰酚-3-O-β-D-葡萄糖醛酸苷和迷迭香酸的含量（n=3）测定结果分别为0.29~1.09、0.03~0.29、0.62~1.37、0.38~1.32、0.06~0.42、0.03~0.51、0.17~0.68、0.05~0.59、0.44~2.22 mg·g^-1。结论：该方法准确可靠，适用于草珊瑚中9个有效成分的含量测定。

Objective: To develop an HPLC method for simultaneous determination of nine active components including astilbin, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, quercetin-3-O-β-D-glucuronide, isofraxidin, kaempferol-3-O-β-D-glucuronide and rosmarinic acid in Sarcandra glabra. Methods: The Diamonsil C₁₈(250 mm×4.6 mm, 5 μm)column was used. Acetonitrile(A)-0.2% phosphoric acid(B)was used as the mobile phase with gradient elution(0-20 min, 2% A→15% A;20-23 min, 15% A→12.5% A;23-42 min, 12.5% A;42-70 min, 12.5% A→30% A), at a flow rate of 1.0 mL·min^-1. The detection wavelength was 290 nm for detecting astilbin and 344 nm for detecting neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, quercetin-3-O-β-D-glucuronide, isofraxidin, kaempferol-3-O-β-D-glucuronide and rosmarinic acid. The column temperature was 35 ℃. Results: The calibration curves of astilbin, neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, quercetin-3-O-β-D-glucuronide, isofraxidin, kaempferol-3-O-β-D-glucuronide and rosmarinic acid were in good linearity over 0.041-2.050, 0.036 0-1.780, 0.046-2.282, 0.047-2.330, 0.040-2.010, 0.021-1.060, 0.047-2.335, 0.024-1.220, 0.047-2.368μg(r=0.999 8-1.000 0), respectively. The average recoveries(n=6)were in the ranges of 98.4%~101.7% and RSD were in the ranges of 1.1%~2.7%. The contents(n=3)in 17 batches of samples of Sarcandra glabra were 0.29-1.09 mg·g^-1 for astilbin, 0.03-0.29 mg·g^-1 for neochlorogenic acid, 0.62-1.37 mg·g^-1 for chlorogenic acid, 0.38-1.32 mg·g^-1 for cryptochlorogenic acid, 0.06-0.42 mg·g^-1 for caffeic acid, 0.03-0.51 mg·g^-1 for quercetin-3-O-β-D-glucuronide, 0.17-0.68 mg·g^-1 for isofraxidin, 0.05-0.59 mg·g^-1 for kaempferol-3-O-β-D-glucuronide, 0.44-2.22 mg·g^-1 for rosmarinic acid. Conclusion: The method is precise and reliable, and can be used to determine the contents of nine effective components in Sarcandra glabra.