黄晓婧;李婷婷;肖春霞;许莉;张定堃;文永盛;谭鹏;
HUANG Xiao-jing;LI Ting-ting;XIAO Chun-xia;XU Li;ZHANG Ding-kun;WEN Yong-sheng;TAN Peng;Chengdu Institute for Food and Drug Control;College of Pharmacy,Chengdu University of Traditional Chinese Medicine;

• 1:成都市食品药品检验研究院
• 2:成都中医药大学药学院

摘要(Abstract)：

为了更全面地评价苏合香的质量,该文尝试建立一种HPLC波长切换法同时测定苏合香中7种成分的含量,通过化学计量学分析不同来源苏合香的质量差异,最终建立苏合香的色谱特征图谱。色谱柱为Agilent ZORBAX Extend C18(4. 6 mm×250 mm,5μm),流动相为乙腈-0. 1%磷酸水溶液,梯度洗脱,检测波长为:0～13. 5 min,194 nm(苯甲酸); 13. 5～20. 5 min,278 nm(肉桂酸); 20. 5～32 min,194 nm(苯甲酸苄酯、肉桂酸苄酯、肉桂酸肉桂酯、脱氢松香酸); 32～55 min,241 nm(松香酸)。方法学验证结果显示苯甲酸、肉桂酸、苯甲酸苄酯、肉桂酸苄酯、肉桂酸肉桂酯、脱氢松香酸、松香酸的进样质量分别在0. 006 948～0. 694 8,0. 001 426～0. 142 6,0. 013 16～0. 658 0,0. 006 148～0. 614 8,0. 008 035～0. 803 5,0. 002 121～0. 212 1,0. 010 172～1. 017 2μg与峰面积呈良好的线性关系,平均加样回收率在94. 34%～105. 8%,RSD均小于3. 0%(n=6)。方法学验证结果表明该文建立的HPLC波长切换法具有较好的准确性、重复性。样品测定结果显示不同来源的苏合香药材质量差异较大。以中国食品药品检定研究院提供的苏合香对照药材(S1)的色谱图为参照谱图,计算各批次样品的指纹图谱相似性。结果显示,S2～S10的相似度分别为0. 952,0. 949,0. 981,0. 351,0. 751,0. 969,0. 979,0. 992,0. 971,0. 971,除S5,S6药材的相似度差异较大外,其他批次的供试品的相似度较好,表明其质量差异较小; S11～S20的相似度分别为0. 060,0. 055,0. 054,0. 285,0. 092,0. 002,0. 044,0. 044,0. 044,0. 040,结果表明S11～S20与对照药材相比,整体质量差异较大。以S1～S10的色谱图为基础,建立了苏合香的HPLC特征图谱,供其他药学研究者参考。该文建立的含量测定方法及特征图谱分析方法简便、重复性好、专属性强,为苏合香的整体质量评控提供了一种参考方法。
In order to evaluate the quality of Styrax more comprehensively,this study attempted to establish an HPLC wavelength switching method to simultaneously determine the content of seven compounds in Styrax,and chemometrics were used to analyze the quality difference between different sources of Styrax,and finally establish a characteristic chromatogram of Styrax. The column was Agilent ZORBAX Extend C18( 4. 6 mm×250 mm,5 μm) with phase a mixture of acetonitrile-0. 1% phosphoric acid aqueous solution as the mobile phase in a gradient elution procedure; the detection wavelength was set as follows: 0-13. 5 min,194 nm( benzoic acid);13. 5-20. 5 min,278 nm( cinnamic acid); 20. 5-32 min,194 nm( benzyl benzoate,benzyl cinnamate,cinnamyl cinnamate,dehydroabietic acid); 32-55 min,241 nm( abietic acid). The methodological verification results showed that when the injection masses of benzoic acid,cinnamic acid,benzyl benzoate,benzyl cinnamate,cinnamyl cinnamate,dehydroabietic acid and abietic acid were0. 006 948-0. 694 8,0. 001 426-0. 142 6,0. 013 16-0. 658 0,0. 006 148-0. 614 8,0. 008 035-0. 803 5,0. 002 121-0. 212 1,and0. 010 172-1. 017 2 μg,respectively,there were good linear relationship between injection mass and peak area. The average recovery rates of seven compounds were in the range from 94. 34% to 105. 8%,and all RSD were less than 3. 0%( n = 6). The methodological verification results showed that the developed HPLC wavelength switching method has good accuracy and repeatability. The results of the sample analysis showed that the quality of Styrax from different sources was quite different. The chromatogram of Styrax reference material( S1) was used as the reference chromatogram to calculate the fingerprint similarity of each batch of samples. The results showed that the similarities of samples S2-S10 were 0. 952,0. 949,0. 981,0. 351,0. 751,0. 969,0. 979,0. 992 and 0. 971,respectively.The similarity values of other batches samples were satisfactory,except for sample S5 and S6,indicating that the quality difference among these samples is small. The similarity values of S11-S20 were 0. 060,0. 055,0. 054,0. 285,0. 092,0. 002,0. 044,0. 044,0. 044,and 0. 040,respectively. The results showed that compared with the sample S1,there was a large quality difference among S11-S20. Based on the chromatograms of S1-S10,the HPLC characteristic chromatograms of Styrax was established and the purpose is to give reference to other pharmaceutical researchers. The newly developed HPLC wavelength switching method have the advantages of simplicity,reproducibility and specificity,and the developed HPLC characteristic chromatograms provided a reference method for the overall quality evaluation of Styrax.

关键词(KeyWords)： 苏合香;HPLC;波长切换法;化学计量学;指纹图谱;质量评价
Styrax;HPLC;wavelength switching;chemometrics;fingerprint chromatogram;quality evaluation

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81873232);; 四川省科技厅应用基础研究项目(2019YJ0640);; 成都市科技局技术创新研发项目(2018-YF05-01277-SN);成都市科技局科技惠民项目(2016-HM01-00312-SF)

作者(Author): 黄晓婧;李婷婷;肖春霞;许莉;张定堃;文永盛;谭鹏;
HUANG Xiao-jing;LI Ting-ting;XIAO Chun-xia;XU Li;ZHANG Ding-kun;WEN Yong-sheng;TAN Peng;Chengdu Institute for Food and Drug Control;College of Pharmacy,Chengdu University of Traditional Chinese Medicine;

Email:

DOI: 10.19540/j.cnki.cjcmm.20190423.201

参考文献(References)：

• [1]中国药典.一部[S]. 2015:164.
• [2]袁开惠,王兴伊.苏合香考[J].中医药文化,2017,12(2):37.
• [3]莫志贤,张宏伟.广东产苏合香对大鼠血液流变学的影响[J].中药材,2003,26(11):804.
• [4]倪彩霞,曾南,苟玲,等.芳香开窍药对脑缺血再灌注损伤大鼠的保护作用及其机制初探[J].中药药理与临床,2010,26(5):64.
• [5]倪彩霞,曾南,苟玲,等.芳香开窍药对脑缺血再灌注损伤大鼠保护作用机制的研究[J].中药药理与临床,2011,27(5):65.
• [6]赵培,岳少乾,马妍,等.苏合香对脑微血管内皮细胞氧糖剥夺/复氧损伤的影响[J].中国临床药理学杂志,2016,32(9):847.
• [7] Koo B S,Lee S I,Ha J H,et al. Inhibitory effects of the essential oil from SuHeXiang Wan on the central nervous system after inhalation[J]. Biol Pharm Bull,2004,27(4):515.
• [8] Cho M K,Kim S H,Hwang D S,et al. Neuroprotective effect of SuHeXiang Wan in Drosophila models of Alzheimer's disease[J]. J Ethnopharmacol,2011,134(3):1028.
• [9] Park S H,Lee S,Hong Y K,et al. Suppressive effects of SuHeXiang Wan on amyloid-β42-induced extracellular signal-regulated kinase hyperactivation and glial cell proliferation in a transgenic Drosophila model of Alzheimer's disease[J]. Biol Pharm Bull,2013,36(3):390.
• [10] Jeon S,Hur J,Jeong H J,et al. SuHeXiang Wan essential oil alleviates amyloid beta induced memory impairment through inhibition of tau protein phosphorylation in mice[J]. Am J Chin Med,2011,39(5):917.
• [11]王洋,徐珠屏,王建,等.苏合香概述[J].中药与临床,2013,4(3):49.
• [12]罗光明,龚千峰,刘贤旺.苏合香研究进展[J].江西中医学院学报,1997,9(1):44.
• [13]闵祺,王闯.气相色谱法同时测定苏合香丸中龙脑、丁香酚及麝香酮的含量[J].药物分析杂志,2018,38(2):359.
• [14]彭颖,夏厚林,周颖,等.苏合香不同提取方法的气相色谱-质谱研究[J].时珍国医国药,2012,23(7):1675.
• [15]中国药典.四部[S]. 2015:374.
• [16]荆文光,张权,邓哲,等.指纹图谱、多成分定量与化学计量学相结合的厚朴药材质量评价[J].中国中药杂志,2019,44(5):975.
• [17]纪鑫,刘晓谦,肖苏萍,等.女贞子中非三萜类成分的定性定量研究[J].中国中药杂志,2019,44(8):1615.
• [18]梁彩霞,杨岚,张东,等.吴茱萸UPLC指纹图谱的建立及质量评价[J].中国实验方剂学杂志,2017,23(14):38.
• [19]陈斐冷,胡正芳,戚进.基于HPLC-UV-CL联用分析技术评价不同产地山麦冬清除过氧化氢活性[J].中国中药杂志,2019,44(5):990.
• [20]何晶,陈敏,陈永刚,等.麦芽发芽过程中HPLC指纹图谱及生物碱含量变化规律[J].中国实验方剂学杂志,2017,23(23):46.
• [21]孙国祥,胡玥珊,智雪枝.用复杂性科学原理揭示中药指纹图谱的本质特征[J].中南药学,2008,6(5):600.
• [22]姚令文,刘燕,郑笑为,等.指纹图谱、特征图谱技术在中药材和中成药中的应用[J].中国新药杂志,2018,27(8):934.