高效液相色谱法同时测定遍山红中8种化学成分含量

Simultaneous determination of eight chemical components in Oxyspora paniculata(Don) DC. using HPLC method

    摘要
  • 摘要: 【目的】建立高效液相色谱法(HPLC)测定遍山红中没食子酸、二氢杨梅素、原儿茶酸、芦丁、杨梅苷、齐墩果酸、槲皮素、山柰酚化学成分含量的方法,以期更好地控制遍山红的药材质量,为其药用开发提供科学依据。【方法】采用ZORBAX SB-C18色谱柱、乙腈-0.2%磷酸为流动相、流速0.6 mL/min、柱温28℃的HPLC测定广西都安县4个采集点(弄苍队、卜凡队、弄外队、百外队)遍山红中8种化学成分的含量。【结果】260 nm是测定混合对照品溶液和遍山红供试液中8种化学成分的最大波长。4个采集点遍山红中8种化学成分的含量范围:没食子酸1.53~1.75 mg/g、原儿茶酸1.11~1.28 mg/g、二氢杨梅素0.99~1.44 mg/g、芦丁0.34~0.42 mg/g、杨梅苷1.58~1.83 mg/g、齐墩果酸0.63~0.77 mg/g、槲皮素1.03~1.11 mg/g、山柰酚0.65~0.69 mg/g。8种化学成分质量浓度与峰面积线性关系及色谱峰分离效果均良好(r>0.9990);没食子酸、原儿茶酸、二氢杨梅素、芦丁、杨梅苷、齐墩果酸、槲皮素和山柰酚的进样量范围分别为7.20~43.20、3.36~20.16、5.92~35.52、1.60~9.60、6.40~38.40、3.68~22.08、3.20~19.20和3.52~21.12μg/mL,对应的平均加样回收率为101.72%、96.49%、97.04%、99.79%、99.80%、99.44%、101.52%和96.63%。【结论】建立的HPLC具有操作简便、准确可靠、灵敏度高、重现性好等优点,可用于遍山红的质量控制。

     

    Abstract: ObjectiveThe present study was conducted to establish high performance liquid chromatography (HPLC) method for determination of gallic acid, dihydromyricetin, protocatechuic acid, rutin, myricetrin, oleanic acid, quercetin and kaempferol in Oxyspora paniculata(Don) DC. in order to better control the quality and provide scientific basis for its medicinal development. MethodThe eight kinds of chemical components in O. paniculata from four collection sites in Du ’ an county , Guangxi ( Nongcang hamlet , Bofan hamlet , Nongwai hamlet , Baiwai hamlet ) were measured by HPLC method with ZORBAX SB-C18 column, acetonitrile-0.2% H3PO4 mobile phase, 0.6 mL/min flow rate and 28 ℃ column temperature.Result260 nm was the maximum wave lengths for determined of mixed reference substances solution and eight chemical components in O. paniculata tested solution. Contents of the eight chemical components in O. paniculata col-lected from the four collection sites were as follows: gallic acid 1 . 53-1 . 75 mg/g , protocatechuic acid 1.11-1.28 mg/g, dihydromyricetin 0.99-1.44 mg/g, rutin 0.34-0.42 mg/g, myricetrin 1.58-1.83 mg/g, oleanic acid 0.63-0.77 mg/g, quercetin 1.03-1.11 mg/g and kaempferol 0.65-0.69 mg/g. Linear relations between mass concentration and peak area and chromatographic peak separation effects of the eight chemical components were sound(r>0.9990). Sample sizes of the eight chemical components were as follows: gallic acid 7.20-43.20 μg/mL, protocatechuic acid 3.36-20.16 μg/mL, dihy-dromyricetin 5.92-35.52 μg/mL, rutin 1.60-9.60 μg/mL, myricetrin 6.40-38.40 μg/mL, oleanic acid 3.68-22.08 μg/mL, quercetin 3.20-19.20μg/mL and kaempferol 3.52-21.12μg/mL. Sample recovery rates of the eight chemical compo-nents were as follows: gallic acid 101.72%, protocatechuic acid 96.49%, dihydromyricetin 97.04%, rutin 99.79%, myricetrin 99.80%, oleanic acid 99.44%, quercetin 101.52% and kaempferol 96.63%. ConclusionThe HPLC method for O. paniculata is simple, feasible, accurate, reliable, highly sensitive and reproducible, which can be used in quality control of. O. paniculata.

     

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