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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(C_(18))色谱柱(250 mm×4.6 mm,5μm),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、0.047~2.368μg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 establish a HPLC method for the simultaneous determination of acesulfame, chlorogenic acid, chlorogenic acid, cryptogenic acid, caffeic acid, quercetin -3-O-β-D-glucuronide, Pyridine, kaempferol-3-O-β-D-glucuronide and rosmarinic acid. METHODS: Diamonsil octadecylsilane bonded silica gel column (250 mm × 4.6 mm, 5 μm) was used. The mobile phase consisted of acetonitrile (A) - 0.2% phosphoric acid solution (B) and 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% 1.0 mL · min -1, detection wavelength was 290 nm (astilbin), 344 nm (chlorogenic acid, chlorogenic acid, cryptocandrin, caffeic acid, quercetin -3-O- D-glucuronide, isoxabine, kaempferol-3-O-β-D-glucuronide and rosmarinic acid) at a column temperature of 35 ° C. Results: Acesulfame, chlorogenic acid, chlorogenic acid, cryptocholic acid, caffeic acid, quercetin -3-O-β-D-glucuronide, isoxapide, O-β-D-glucuronide and rosmarinic acid were in the range of 0.041-2.050, 0.036-0-1.780, 0.046-2.282, 0.047-2.330, 0.040-2.01,0.021-1.060, 0.047-2.335, 0.024-1.220, The linear range was 0.047 ~ 2.368μg (r = 0.999 8 ~ 1.000). The average recoveries (n = 6) ranged from 98.4% to 101.7% with RSD ranging from 1.1% to 2.7%. In a sample of 17 species of grass coral, astilbin, neogenic chlorogenic acid, chlorogenic acid, cryptocholic acid, caffeic acid, quercetin -3-O-β-D-glucuronide, isoxabepil, The content of phenol-3-O-β-D-glucuronide and rosmarinic acid (n = 3) were 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. Conclusion: The method is accurate and reliable and is suitable for the determination of nine effective components in the coral.