新鲜天麻中天麻素与对羟基苯甲醇含量及影响因素分析

Influencing Factors Analysis on the Content of Gastrodin and P-hydroxybenzyl Alcohol in Fresh Gastrodia elata

目的:探究新鲜天麻中天麻素与对羟基苯甲醇含量的影响因素。方法:本研究优化了新鲜天麻中天麻素与对羟基苯甲醇含量测定的前处理方法和色谱条件,样品经匀浆、87%乙醇混匀、超声提取、离心、氮吹结合水浴浓缩,过0.45μm滤膜后,在220 nm波长下,采用C 18色谱柱(4.6 mm×250 mm,5μm)、以乙腈-0.05%磷酸水溶液(3∶97,V/V)为流动相进行等度洗脱,流速1.0 mL·min^(-1),柱温25℃,通过高效液相色谱法(HPLC)对遵义市汇川区采集的不同等级大小、不同产地以及不同栽培方式的72份新鲜天麻中天麻素与对羟基苯甲醇含量进行分析测定。结果:方法学验证结果表明,在3个加标浓度下,天麻素、对羟基苯甲醇2种成分在相应范围内线性关系良好,R^(2)>0.9999,回收率在81.20%~107.30%之间,RSD<5%,得到所测72份样品中天麻素与对羟基苯甲醇总含量范围为0.262%~1.150%。本研究使用的鲜天麻前处理制样方法简便、快速且提取效果良好,分析检测方法具有良好的精密度、重现性和分离效果,适用于新鲜天麻中天麻素与对羟基苯甲醇含量的研究分析。通过进一步分析不同等级大小、不同产地以及不同栽培方式对新鲜天麻中天麻素与对羟基苯甲醇含量影响,结果表明鲜天麻的质量等级越大,天麻素与对羟基苯甲醇总含量越高;由于各天麻种植基地所在的海拔高低、环境植被种类、地理位置、光照等存在差异,且每个种植基地林下种植天麻的发展时间有所不同,因此,不同产地的鲜天麻中天麻素与对羟基苯甲醇含量也存在显著差异;而对不同栽培方式的鲜天麻进行比较分析,发现有性繁殖优于无性繁殖。结论:汇川区不同产地、不同栽培方式及不同等级大小的新鲜天麻中天麻素与对羟基苯甲醇含量的影响具有一定的差异性。

Objective:To explore the factors affecting the content of gastrodin and p-hydroxybenzyl alcohol in fresh Gastrodia elata.Methods:This study optimized the pre-treatment sample preparation method and chromatographic conditions for the determination of gastrodin and p-hydroxybenzyl alcohol in fresh Gastrodia elata.The sample was homogenized,mixed with 87%ethanol,ultrasonic extraction,centrifugation,nitrogen blowing and water bath concentration.After passing through a 0.45μm filter membrane,the sample was eluted equivalently on a C 18 column(4.6 mm×250 mm,5μm)with acetonitrile-0.05%phosphoric acid aqueous solution(3∶97,V/V)as the mobile phase at 220 nm.The flow rate was 1.0 mL·min^(-1) and the column temperature was 25℃,high performance liquid chromatography(HPLC)was used to analyze and determine the contents of gastrodin and p-hydroxybenzyl alcohol in 72 fresh Gastrodia elata collected from different size hierarchy,origins and cultivation methods in Huichuan district,Zunyi city.Results:The results of methodology validation showed that the linear relationship of gastrodin and p-hydroxybenzyl alcohol was good in the corresponding range under three spiked concentrations,R^(2)>0.9999,the recovery was 81.20%~107.30%,RSD<5%.The total content of gastrodin and p-hydroxybenzyl alcohol in 72 samples was 0.262%~1.150%.The sample preparation method of fresh gastrodia elata used in this study is simple,rapid and has good extraction effect.The analytical method has good precision,reproducibility and separation effect,which is suitable for the study and analysis of gastrodin and p-hydroxybenzyl alcohol in fresh gastrodia elata.Through further analysis of the effects of different grades,sizes,origins,and different cultivation methods on the content of gastrodin and p-hydroxybenzyl alcohol in fresh gastrodia,the results showed that the higher the quality grade of fresh gastrodia,the higher the total content of gastrodin and p-hydroxybenzyl alcohol;Gastrodia planting bases are located at different altitudes,environmental vegetation types,geographic locations,and sunlight,and the development time of Gastrodia planting under forests in each planting base is different.Therefore,the gastrodin and p-hydroxy There are also significant differences in the content of benzyl alcohol;and a comparative analysis of fresh Gastrodia elata with different cultivation methods reveals that sexual reproduction is better than asexual reproduction.Conclusion:The influence of gastrodin and p-hydroxybenzyl alcohol in fresh Gastrodia elata.in different origins,cultivation methods and size hierarchy in Huichuan district has certain differences.