摘要
目的:建立黄精及其饮片中薯蓣皂苷元、5-羟甲基糠醛、香草酸、芦丁、槲皮素及山柰酚6种成分的一测多评(QAMS)法。方法:采用高效液相色谱(HPLC)外标法同时测定黄精及其饮片中6种成分的含量[色谱柱为Diamonsil-C_(18);流动相为乙腈-水(梯度洗脱);5-羟甲基糠醛、香草酸、芦丁、槲皮素及山柰酚检测波长为254 nm(0～60 min),薯蓣皂苷元检测波长为202 nm(60～75min);流速为1.0 mL/min;柱温为30℃]。以香草酸为内标,分别计算出其余5种成分的相对校正因子(RCFs)并进行耐用性考察,并采用相对保留法对待测成分色谱峰进行准确定位,然后根据RCFs计算黄精药材中其他5种成分的含量,并与外标法测定结果进行比较。以黄精饮片进行方法验证,分别采用QAMS法和外标法测定其中6种成分的含量,比较两种方法含量测定差异。结果:HPLC方法学考察结果均符合相关要求。在线性范围内,薯蓣皂苷元、5-羟甲基糠醛、芦丁、槲皮素及山柰酚的RCFs分别为0.195、0.025、0.263、0.345、0.075,在不同试验条件下RCFs重现性良好。外标法和QAMS法两种方法测得的黄精药材及其饮片中6种成分的含量差异均无统计学意义(P>0.05)。结论:建立的QAMS法可用于同时测定黄精药材及其饮片中6种成分含量。
OBJECTIVE:To establish a quantitative analysis of multi-components by singer marker(QAMS)for determining the contents of diosgenin,5-hydroxymethylfurfural,vanillic acid,rutin,quercetin,kaempferol in Polygonati Rhizoma and its decoction piece. METHODS:HPLC external standard method was used to determine the contents of 6 components in Polygonati Rhizoma and its decoction piece simultaneously [the separation was carried out on Diamonsil-C_(18) column with mobile phase consisted of acetonitrile-water(gradient elution);the detection wavelengths of 5-hydroxymethylfurfural,vanillic acid,rutin,quercetin and kaempferol were set at 254 nm(0-60 min);the detection wavelength of diosgenin was set at 202 nm(60-75 min)at the flow rate of 1.0 mL/min. The column temperature was 30 ℃ ]. Using vanillic acid as internal standard,relative correction factors(RCFs) of aother 5 components were calculated and to investigate durability. Relative retention method was used to accurately locate the chromatographic peaks of the components to be determined,and then the contents of the aother 5 components in Polygonati Rhizoma were calculated according to RCFs,and the results were compared with those determined by external standard method. The method was validated by Polygonati Rhizoma decoction piece. The contents of 6 components were determined by QAMS method and external standard method respectively,and then the differences of content determination were compared between 2 methods. RESULTS:The methodology investigation results of HPLC method were in line with related requirements.Within the linear range,the RCFs of diosgenin,5-hydroxymethylfurfural,rutin,quercetin,kaempferol were 0.195,0.025,0.263,0.345 and 0.075,respectively. Under different experiment conditions,RCFs showed good reproducibility;there was no statistical significance of 6 components in Polygonati Rhizoma and its decoction piece determined by external standard method and QAMS method(P>0.05). CONCLUSIONS:Established QAMS method is suitable for simultaneous determination of 6 components in Polygonati Rhizoma and its decoction piece.
引文
[1]国家药典委员会.中华人民共和国药典:一部[S].2015年版.北京:中国医药科技出版社,2015:306-307.
[2]陶爱恩,张晓灿,杜泽飞,等.黄精属植物中黄酮类化合物及药理活性研究进展[J].中草药,2018,49(9):2163-2171.
[3]王婷,苗明三.黄精的化学、药理及临床应用特点分析[J].中医学报,2015,30(5):714-715.
[4]庞玉新,赵致,袁媛,等.黄精的化学成分及药理作用[J].山地农业生物学报,2003,22(6):547-550.
[5]SETHI G,SHANMUGAM MK,WARRIER S,et al.Proapoptotic and anti-cancer properties of diosgenin:a comprehensive and critical review[J].Nutrients,2018,10(5):645-672.
[6]TANER G,OZKAN VARDAR D,AYDIN S,et al.Use of in vitro assays to assess the potential cytotoxic,genotoxic and antigenotoxic effects of vanillic and cinnamic acid[J].Drug Chem Toxicol,2017,40(2):183-190.
[7]张洪,张惟郁,朱伟嵘.黄酮类化合物调控肿瘤细胞自噬的研究进展[J].中草药,2017,48(24):5252-5262.
[8]钟凌云,龚千峰,张的凤,等.黄精炮制研究现状分析[J].中药材,2007,30(12):1618-1621.
[9]秦宇雯,袁玮,陆兔林,等.九华黄精的炮制工艺沿革及现代研究[J].中草药,2018,49(18):4432-4438.
[10]魏征,曾林燕,宋志前,等.顶空-气相色谱-质谱联用分析黄精炮制过程化学成分的变化[J].中国实验方剂学杂志,2012,18(20):115-118.
[11]常亮,陈珍珍,王栋,等.HPLC和GC-MS法测定三种黄精炮制过程中5-羟甲基糠醛的含量[J].中国药师,2015,18(3):387-390.
[12]王智民,钱忠直,张启伟,等.一测多评法建立的技术指南[J].中国中药杂志,2011,36(6):657-660.
[13]卿大双,罗维早,孙建彬,等.一测多评法测定黄连及其炮制品中6种生物碱[J].中草药,2016,47(2):324-329.
[14]杨菲,王智民,张启伟,等.“一测多评”法测定丹参酚酸类成分的含量[J].中国中药杂志,2011,36(17):2372-2379.
[15]冯伟红,李春,张锴镔,等.“一测多评”模式在淫羊藿药材、饮片及含淫羊藿中成药黄酮类成分检测中的一体化研究[J].中国中药杂志,2016,41(15):2843-2854.
[16]程俊,余佩佩,顾华,等.高效液相色谱-二极管阵列/荧光检测器串联法同时测定化妆品中8种性激素[J].分析试验室,2016,35(1):112-116.
[17]宋艺君,郭涛,周晓程.不同产地黄精经不同方法炮制后多糖、5-羟甲基糠醛的含量变化[J].中国药房,2017,28(16):2256-2258.
[18]马福成,马小龙,岳丽君.含黄精药物中5-羟甲基糠醛、糠醛及总黄酮同时测定的反相高效液相色谱方法研究[J].国际药学研究杂志,2018,45(6):455-459.
[19]陈辉,冯珊珊,孙彦君,等.3种药用黄精的化学成分及药理活性研究进展[J].中草药,2015,46(15):2329-2338.
[20]王晶晶,梁娟娟,陈两绵,等.一测多评法结合UPLC-Q-TOF/MS评价天舒胶囊中主要苯酞类成分[J].中国实验方剂学杂志,2015,21(22):86-93.
