目的: 建立超高效液相色谱-三重四极杆质谱（UPLC-QQQ MS/MS）法测定茜草及其炮制品中遗传毒性物质芦西定和芦西定-3-O-素草苷的含量。方法: 采用Waters ACQUITY UPLC BEH C18（100 mm×2.1 mm,1.7 μm）色谱柱,以0.1%甲酸-乙腈为流动相,梯度洗脱,体积流量0.3 mL · min-1,柱温30 ℃,进样量为1 μL。采用质谱检测器,离子化模式为ESI+,多反应监测。同时以聚类分析和偏最小二乘判别分析模型对不同规格药材结果进行分析。结果: 芦西定及其芦西定-3-O-素草苷在各自范围内线性关系良好（r>0.990 0）,平均加样回收率为99.8%和98.3%,RSD为1.1%和1.2%。生品中芦西定含量为6.51~26.28 μg · g-1,芦西定-3-O-素草苷含量为85.77~232.73 μg · g-1;炮制不到位茜草炭中芦西定含量为5.13~12.45 μg · g-1,芦西定-3-O-素草苷含量为33.31~196.52 μg · g-1;炮制到位茜草炭中芦西定含量为15.78~40.85 μg · g-1,芦西定-3-O-素草苷含量为0~16.12 μg · g-1。结论: 建立了茜草中芦西定和芦西定-3-O-素草苷的含量测定方法,且该方法准确稳定,重复性好,耐用性强,可以用于含量检测。炮制炒炭后遗传毒性物质芦西定和芦西定-3-O-素草苷的含量有明显差异。
江凯航
,
张明童
,
黎志兵
,
冯学英
,
刘志浩
,
李子彬
,
刘东升
,
李冬华
,
马潇
. 基于UPLC-QQQ MS/MS测定茜草及其炮制品中遗传毒性物质芦西定和芦西定-3-O-素草苷的含量*[J]. 药物分析杂志, 2025
, 45(4)
: 679
-685
.
DOI: 10.16155/j.0254-1793.2024-1089
Objective: To establish an ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QQQ MS/MS) method for the determination of genotoxic lucidin and lucidin-3-O-primeveroside in raw and processed Rubiae Radix et Rhizoma. Methods: The analysis was performed on a Waters ACQUITY UPLC BEH C18 column (100 mm×2.1 mm, 1.7 μm) using 0.1% formic acid-acetonitrile as the mobile phase in gradient elution mode. The flow rate was set at 0.3 mL · min-1, the column temperature was maintained at 30 ℃, and the injection volume was 1 μL. Detection was carried out by mass spectrometry with ESI+ ionization mode and multiple reaction monitoring (MRM). Additionally, the results from different grades of medicinal materials were analyzed using cluster analysis and partial least squares-discriminant analysis (PLS-DA) models. Results: Lucidin and lucidin 3-O-beta-primveroside were found to exhibit good linearity (r>0.990 0) within their respective concentration ranges. The mean recovery rates were determined to be 99.8% and 98.3% with RSDs of 1.1% and 1.2%, respectively. Content of lucidin in the prepared slices were 6.51-26.28 μg · g-1, while those of lucidin-3-O-primeveroside were 85.77-232.73 μg · g-1. Contents of lucidin and lucidin-3-O-primeveroside in the incompletely processed Rubiae Radix et Rhizoma charcoal were 5.13-12.45 μg · g-1, 33.31-196.52 μg · g-1.In properly processed Rubiae Radix et Rhizoma charcoal, content of lucidin were 15.78-40.85 μg · g-1, whereas those of lucidin-3-O-primeveroside was reduced to undetectable or trace levels (0-16.12 μg · g-1). Conclusion: This study establishes a reliable method for quantifying lucidin and lucidin-3-O-primeveroside in Rubiae Radix et Rhizoma. The method demonstrates excellent accuracy, stability, reproducibility, and robustness, making it suitable for routine content analysis. The charcoal-processing (stir-frying) procedure significantly alters the content levels of these genotoxic substances.
[1] 中华人民共和国药典2020年版.一部[S].2020:245
ChP 2020.Vol Ⅰ[S].2020:245
[2] 张凌云. 茜草与混伪品的鉴别[J].时珍国医国药,2006,17(11):2178
ZHANG LY.Identification of Rubiae Radix et Rhizoma and its adulterants[J].Lishizhen Med Mater Med Res,2006,17(11):2178
[3] 王珺,刘亚蓉,宋霞,等.基于多元统计分析对茜草与藏茜草特征图谱的研究[J].药物分析杂志,2022,42(11):2043
WANG J,LIU YR,SONG X,et al.Study on characteristic fingerprints of Rubiae Radix et Rhizoma and Rubia Wallichiana Radix et Rhizoma based on multivariate statistical analysis[J].Chin J Pharm Anal,2022,42(11):2043
[4] 梁锡锋. 诗经[M].开封:河南大学出版社,2008:142
LIANG XF.Shijing[M].Kaifeng:Henan University Press,2008:142
[5] 吴普. 神农本草经[M].孙星衍,孙冯翼,辑.北京:科学技术文献出版社,1996:28
WU P.Shennong Bencao Jing[M].SUN XY,SUN FY,edit.Beijing:Science and Technology Literature Press,1996:28
[6] 李慧,孟宪生.中药茜草的本草考证[J].中药材,2018,41(10):2462
LI H,MENG XS.Textual research on the herbal origin of Rubiae Radix et Rhizoma in traditional Chinese medicine[J].J Chin Med Mater,2018,41(10):2462
[7] 张琳,胡本祥,安衍茹,等.茜草本草考证[J].吉林中医药,2018,38(3):317
ZHANG L,HU BX,AN YR,et al.Herbalogical study on Rubiae Radix et Rhizoma[J].Jilin J Tradit Chin Med,2018,38(3):317
[8] 陈毅,王海丽,薛露,等.茜草的研究进展[J].中草药,2017,48(13):2771
CHEN Y,WANG HL,XUE L,et al.Research progress on Rubiae Radix et Rhizoma[J].Chin Tradit Herb Med,2017,48(13):2771
[9] 李欣,高天鹏,张明童,等.基于多组分含量测定及多元统计分析研究茜草、大茜草及大叶茜草[J].药物分析杂志,2023,43(12):2072
LI X,GAO TP,ZHANG MT,et al.Based on multi-component content determination and multivariate statistical analysis study Rubiae Radix et Rhizoma,Rubia magna P.G.Hsiao.and Rubia schumanniana Pritz.[J].Chin J Pharm Anal,2023,43(12):2072
[10] POGINSKY B,WESTENDORF J,BLÖMEKE B,et al.Evaluation of DNA-binding activity of hydroxyanthraquinones occurring in Rubia tinctorum L[J].Carcinogenesis,1991,12(7):1265
[11] 闫明,王雪,秦超,等.光泽汀体内遗传毒性风险评价[J].药物评价研究,2022,45(7):1248
YAN M,WANG X,QIN C,et al.Risk assessment of in vivo genotoxicity of lucidin[J].Drug Eval Res,2022,45(7):1248
[12] 司南,王宏洁,杨健,等.茜草中Lucidin、Lucidin-3-O-primeveroside及大叶茜草素在小鼠血浆中代谢趋势的初步研究[J].中国实验方剂学杂志,2010,16(2):113
SI N,WANG HJ,YANG J,et al.Preliminary study on the metabolic trends of lucidin,lucidin-3-O-primeveroside,and mollugin in mouse plasma from Rubiae Radix et Rhizoma[J].Chin J Exp Tradit Med Form,2010,16(2):113
[13] 王钢力,田金改,陈德昌.茜草与欧茜草的化学成分研究Ⅱ.反相高效液相色谱法测定茜草素和芦西定的含量[J].药物分析杂志,1997,17(4):219
WANG GL,TIAN JG,CHEN DC.Study on chemical composition of Rubia cordifolia L.and Rubia tinctorum L.Ⅱ.Quantitative determination of alizarin and lucidin by RP-HPLC[J].Chin J Pharm Anal,1997,17(4):219
[14] 司南,杨健,王宏洁,等.液相色谱方法分析茜草中遗传毒性成分Lucidin及其苷[J].中国实验方剂学杂志,2010,16(6):88
SI N,YANG J,WANG HJ,et al.HPLC study on Rubia cordifolia L.contains toxic compositions of lucidin and lucidin-3-O-primeveroside[J].Chin J Exp Tradit Med Form,2010,16(6):88
[15] HENDERSON RL,RAYNER CM,BLACKBURN RS.Isolation and extraction of lucidin primeveroside from Rubia tinctorum L.and crystal structure elucidation[J].Phytochemistry,2013,95:105
