不同采收期贵州乌天麻中三种成分动态分析
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摘要
为促进天麻的驯化栽培及合理采收,以贵州乌天麻为试验材料,采用高效液相色谱-紫外检测(HPLCUV)方法,研究了乌天麻中天麻素(Gastrodin)、对羟基苯甲醇(P-Hydroxybenzylalcohol)、对羟基苯甲醛(PHydroxybenzaldehyde)的含量,并对不同采收期贵州乌天麻中3种成分的动态变化规律进行分析。结果表明:天麻素、对羟基苯甲醇、对羟基苯甲醛的线性范围分别为0.010 16～1.016 00,0.105 2～2.104 0,0.010 21～0.204 20μg(r≥0.999 7),检测限分别为0.67,0.35,0.34ng。定量限分别为2.03,1.05,1.02ng。精密度、稳定性(24h)、重复性试验的RSD%均小于2%(n=6);加标回收率为99.94%～100.54%,RSD为0.98%～1.64%(n=9)。在不同的采收期,贵州乌天麻中3种成分含量有差异,其中天麻素在1月采收时含量较高,对羟基苯甲醇在11月采收含量较高,而对羟基苯甲醛在不同采收期含量变化不大;但不同采收期乌天麻中天麻素与对羟基苯甲醇含量之和均大于0.25%,符合药典规定。所建立的HPLC-UV方法,操作简单,准确度高,可用于贵州乌天麻的质量控制。
In order to promote domestication cultivation and rational harvest of Gastrodiae rhizoma,an HPLCUV method was established for simultaneous determination of gastrodin,p-Hydroxybenzylalcohol and pHydroxybenzaldehyde in Gastrodiae rhizoma harvested on different periods.The detection wavelength was set at 225 nm and the column was maintained at 30℃.10μL solution was injected for analysis.The results showed that the linear ranges for gastrodin,P-Hydroxybenzylalcohol and P-Hydroxybenzaldehyde were 0.010 16-1.016 00,0.105 2-2.104 0 and 0.010 21～0.204 20μg.The limits of detection were 0.67,0.35,0.34 ng,and the limits of quantification were 2.03,1.05,1.02 ng,respectively.The relative standard deviations(RSDs)of precision,stability(24 h),and reproducibility tests for all the three components were lower than 2.0%(n=6).The recovery test for accuracy was conducted and the results were 99.94%-100.54%(RSD=0.98%-1.64%,n=9).The results INDICATed that gastrodin was highest in sample of harvest period in January,while the content of P-Hydroxybenzylalcohol was highest in that of November.The content of P-Hydroxybenzaldehyde was maintained in low level of all harvest periods.However,the sum contents of gastrodin and P-Hydroxybenzylalcohol in all the sample of different harvest periods were higher than 0.25%,which meet the requirement of China Pharmacopoeia.The established HPLC method was simple,reliable and suitable for the quality control of G.rhizoma.
In order to promote domestication cultivation and rational harvest of Gastrodiae rhizoma,an HPLCUV method was established for simultaneous determination of gastrodin,p-Hydroxybenzylalcohol and pHydroxybenzaldehyde in Gastrodiae rhizoma harvested on different periods.The detection wavelength was set at 225 nm and the column was maintained at 30℃.10μL solution was injected for analysis.The results showed that the linear ranges for gastrodin,P-Hydroxybenzylalcohol and P-Hydroxybenzaldehyde were 0.010 16-1.016 00,0.105 2-2.104 0 and 0.010 21～0.204 20μg.The limits of detection were 0.67,0.35,0.34 ng,and the limits of quantification were 2.03,1.05,1.02 ng,respectively.The relative standard deviations(RSDs)of precision,stability(24 h),and reproducibility tests for all the three components were lower than 2.0%(n=6).The recovery test for accuracy was conducted and the results were 99.94%-100.54%(RSD=0.98%-1.64%,n=9).The results INDICATed that gastrodin was highest in sample of harvest period in January,while the content of P-Hydroxybenzylalcohol was highest in that of November.The content of P-Hydroxybenzaldehyde was maintained in low level of all harvest periods.However,the sum contents of gastrodin and P-Hydroxybenzylalcohol in all the sample of different harvest periods were higher than 0.25%,which meet the requirement of China Pharmacopoeia.The established HPLC method was simple,reliable and suitable for the quality control of G.rhizoma.
引文
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[14]闫宝庆,张晖芬,逄楠楠,等.RP-HPLC同时测定天麻中4种成分的含量[J].中国中药杂志,2009,34(22):2903-2906.
[15]周波,张强,汪娟.吉林省长白山区规模化中药材种植产业调查报告[J].黑龙江农业科学,2017(12):76-79.
[16]Tsai C C,Wu K M,Chiang T Y,et al.Comparative transcriptome analysis of Gastrodia elata(Orchidaceae)in response to fungus symbiosis to identify gastrodin biosynthesis-related genes[J].BMC Genomics,2016,17(1):212.
[2]国家药典委员会.中华人民共和国药典[M].北京:中国医药科技出版社.2015.
[3]程立君,罗家刚,胡志芳,等.四个天麻品种种植效果的对比分析[J].北方园艺,2016(20):156-158.
[4]刘威,赵致,王华磊,等.不同树种菌材对贵州仿野生栽培天麻的影响[J].北方园艺,2015(10):129-132.
[5]周楠楠,朱燃,赵雪梅,等.天麻素抑制小鼠大脑内Aβ斑块形成及其作用机制[J].药学学报,2016,51(4):588-594.
[6]郑琴,刘丹,胡鹏翼,等.P-糖蛋白及多药耐药相关蛋白1抑制剂促进天麻素跨膜转运作用机制研究[J].中草药,2016,47(21):3840-3847.
[7]彭正午.天麻素的神经保护作用及其分子机制研究[D].西安:第四军医大学,2016.
[8]Hsieh M T,Wu C R,Chen C F.Gastrodin and p-hydroxybenzyl alcohol facilitate memory consolidation and retrieval,but not acquisition,on the passive avoidance task in rats[J].Journal of Ethnopharmacology,1997,56(1):45-54.
[9]Sung J A,Seung K P,Koo H I,et al.Gastrodin decreases immunoreactivities ofγ-aminobutyric acid shunt enzymes in the hippocampus of seizure-sensitive gerbils[J].Journal of Neuroscience Research,2003,71(4):534-543.
[10]游金辉,谭天秩,匡安仁,等.~3H-天麻甙元和~3H-天麻素在小鼠体内的分布和代谢[J].华西医科大学学报,1994(3):325-328.
[11]马风伟,杨兴成,王瑞,等.天麻中巴利森苷类化合物的结构解析研究进展[J].贵阳学院学报(自然科学版),2018,13(3):76-83.
[12]郭营营,蒋石,林青,等.天麻中对羟基苯甲醇抗血小板聚集的作用及机制研究[J].时珍国医国药,2014,25(1):4-6.
[13]肖远灿,董琦,迟晓峰,等.HPLC同时测定西藏栽培天麻中天麻素和8种核苷及碱基类成分[J].中国中药杂志,2014,39(19):3798-3802.
[14]闫宝庆,张晖芬,逄楠楠,等.RP-HPLC同时测定天麻中4种成分的含量[J].中国中药杂志,2009,34(22):2903-2906.
[15]周波,张强,汪娟.吉林省长白山区规模化中药材种植产业调查报告[J].黑龙江农业科学,2017(12):76-79.
[16]Tsai C C,Wu K M,Chiang T Y,et al.Comparative transcriptome analysis of Gastrodia elata(Orchidaceae)in response to fungus symbiosis to identify gastrodin biosynthesis-related genes[J].BMC Genomics,2016,17(1):212.