A HPLC method for determination of the contents of six components in raw Gastrodia elata,such as gastrodin,p-hydroxybenzyl alcohol,p-hydroxybenz aldehyde,adenosine,parishin A and 4,4'-dihydroxydibenzyl ether,was presented.The analysis was performed on an Agilent 1120 HPLC system with YMC-PEAK ODS-A column (250 mm × 4.6 mm,5 μm).The six components were separated in 80 mins with gradient elution using mobile phase composed of 0.1％ CH3COOH-H2O (A) and CH3OH (B):0～5 min,5％ B;5～65 min,5％～40％ B;65～80 min,40％～ 100％ B.The flow rate of carrier gas was set at 1 mL/min,the column temperature was set at 25 ℃,and the injected sample volume was 50 μL.Six ingredients showed a good linear relationship (R≥0.999 2) within the linear range,and the average recovery rates were in the range of 94.20％～99.68％,and the relative standard deviation was less than 1.96.There were differences in the amounts of six ingredients among different varieties of raw Gastrodia elata,for instance,the contents of the six ingredients in Gastrodia elata f.elata were slightly higher than those in Gastrodia elata f.glauca.In the same variety,the content of parishin glycosides was higher while the contents of adenosine and 4,4'-dihydroxydibenzyl ether were lower.The six ingredients such as gastrodin extracted from fresh gastrodin were stable within 8 hours.This method has the advantage of good separating effect and repeatability,rapid and simple,which could provide references for objective and comprehensive researches on the Rhizoma Gastrodiae.%建立了高效液相色谱测定鲜天麻中天麻素、对羟基苯甲醇、对羟基苯甲醛、腺苷、巴利森苷A、4,4'-二羟基二苄基醚6种成分含量的方法.采用Agilent 1120高效液相系统,YMC-PEAK ODS-A column (250 mm×4.6 mm,5μm)色谱柱,流动相为0.1％甲酸水溶液(A)-甲醇溶液(B),梯度洗脱:0～5 min,5％B;5～65 min,5％ ～40％B;65～ 80 min,40％～100％B;分析时间80 mins;流速1 mL/min;柱温25℃;进样量50 μL.其中天麻素、对羟基苯甲醇、对羟基苯甲醛、腺苷、巴利森苷A、4,4'-二羟基二苄基醚6种成分在线性范围内均具有良好的线性关系(R≥0.999 2);平均回收率在94.20％～99.68％,相对标准偏差小于1.96％.在不同品种的鲜天麻中,6种成分的量存在差异,红天麻各成分含量稍高于乌天麻.同一品种天麻中,巴利森苷类成分含量较高,腺苷和4,4'-二羟基二苄基醚含量均较低,且鲜天麻中提取的天麻素等6种成分在8h内稳定.该方法分离效果与重复性好、快速、简便,能够为客观、全面地研究天麻药材提供参考.
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