目的:建立多组分含量测定并结合多元统计分析,研究茜草与大茜草、大叶茜草,以期找出三者的成分差异,为茜草的鉴别提供研究基础。方法:以ACE Excel 5 C18(250 mm×4.6 mm,5 μm)为色谱柱,以甲醇(A)-0.1%磷酸溶液(B)为流动相,进行梯度洗脱,流速为1.0 mL·min-1,检测波长为250 nm,进样量为10 μL,柱温为30 ℃。对24批茜草饮片、10批大茜草和8批大叶茜草进行茜素、芦西定、异茜草素、羟基茜草素、6-羟基甲基异茜草素、大黄酚、大叶茜草素7个成分的含量测定,并将测得结果通过聚类和偏最小二乘法(PLS-DA)进行分析评价。结果:建立了茜草、大茜草与大叶茜草的多组分含量测定方法,测得7个成分的质量分数分别为:0.000 4%~0.037 6%、未检出~0.049 2%、0.001 0%~0.016 9%、0.003 4%~0.052 5%、0.002 8%~0.086 9%、0.002 3%~0.018 6%、0.059 5%~0.298 8%;通过聚类分析与偏最小二乘法(PLS-DA)均可将茜草聚为1类,大茜草聚为1类,大叶茜草聚为1类。结论:通过多组分含量测定,应用聚类分析、偏最小二乘法(PLS-DA)可区分茜草与大茜草、大叶茜草。此方法的建立为茜草的质量控制及评价提供了分析方法和数据支撑。
Objective: To establish the multicomponent content determination and multivariate statistical analysi and study Rubiae Radix et Rhizoma, Rubia magna P.G. Hsiao. and Rubia schumanniana Pritz., in order to find out the difference between the three components, to provide the research basis for the identification of Rubiae Radix et Rhizoma. Methods: The HPLC analysis was carried out on ACE Excel 5 C18 column(250 mm×4.6 mm, 5 μm)with methanol(A)-0.1% phosphoric acid solution(B) was used as the mobile phasa. The flow rate was 1.0 mL·min-1, the detection wavelength was 250 nm and the injection volume was 10 μL, the column temperature was 30 ℃, the contents of alizarin, lucidin, xanthopurpurin, purpurin, 6-hydroxyrubiadin, chrysophanol, mollugin 7 components in 24 batches of Rubiae Radix et Rhizoma, 10 batches of Rubia magna P.G. Hsiao. and 8 batches of Rubia schumanniana Pritz. were determined, the results were evaluated by clustering and partial least square method(PLS-DA). Results: A multi-component content determination method was established, the mass fractions were respectively measured as follows: 0.000 4%-0.037 6%, not detected-0.049 2%, 0.001 0%-0.016 9%, 0.003 4%-0.052 5%, 0.002 8%-0.086 9%, 0.002 3%-0.018 6%, 0.059 5%-0.298 8%. By cluster analysis and partial least squares (PLS-DA), Rubiae Radix et Rhizoma could be grouped into one class, Rubia magna P. G. Hsiao. was grouped into one group, Rubia schumanniana Pritz. was grouped into one group. Conclusion: By multi-component content determination, using cluster analysis and partial least squares (PLS-DA), we can distinguish Rubiae Radix et Rhizoma, Rubia magna P.G. Hsiao., Rubia schumanniana Pritz.. The establishment of this method provides analysis method and data support for the quality control and evaluation of Rubiae Radix et Rhizoma.
[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 Chin Med,2018, 38(3):317
[3] 刘振启, 刘杰. 茜草与混乱品种的鉴别[J]. 首都医药, 2011, 18(21):49
LIU ZQ, LIU J. Identification of Rubiae Radix et Rhizoma and chaotic varieties[J]. Cap Med J, 2011, 18(21):49
[4] 何世新, 张华, 牟腊梅, 等. 4个产地大叶茜草重金属测定及品质评价[J]. 中成药,2021, 43(11):3248
HE SX, ZHANG H, MU LM, et al. Determination of heavy metals and quality evaluation of Madder Rubium L. from 4 producing areas[J]. Chin Tradit Pat Med, 2021, 43(11):3248
[5] 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 finger prints 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
[6] 李建, 黄宇思, 赖慧, 等. ICP-MS法用于不同产地大叶茜草中9种微量元素的含量测定及质量评价[J]. 西北药学杂志, 2022, 37(5):8
LI J, HUANG YS, LAI H, et al. Determination of 9 trace elements and quality evaluation of the root of Rubia schumanniana Pritz. from different origins by ICP-MS[J]. Northwest J Pharm,2022, 37(5):8
[7] 王闽予, 何民友, 李国卫, 等. 不同产地茜草药材UPLC指纹图谱研究及化学计量学分析[J]. 广东药科大学学报, 2020, 36(5):609
WANG MY, HE MY, LI GW, et al. Study on UPLC fingerprint and chemometric analysis of Rubia cordifolia L. from different habitats[J]. J Guangdong Pharm Univ, 2020, 36(5):609
[8] 何燕玲, 汪洋, 马雪, 等. 黔产钩毛茜草HPLC指纹图谱建立及化学模式识别研究[J]. 中药材, 2019, 42(11):2593
HE YL, WANG Y, MA X, et al. Study on the establishment of HPLC fingerprints and chemical pattern recognition of Rubia oncotricha from Guizhou[J]. J Chin Med Mater, 2019, 42(11):2593
[9] 陈一龙, 范刚, 刘悦, 等. 茜草及其混伪品DNA条形码鉴定[J]. 中国药学杂志, 2015, 50(15):1266
CHEN YL, FAN G, LIU Y, et al. DNA barcoding identification of Rubiae Radix et Rhizoma and its adulterants[J]. Chin J Pharm, 2015, 50(15):1266
[10] 李慧, 包永睿, 王帅, 等. 基于多元统计分析的茜草药材指纹图谱研究[J]. 时珍国医国药, 2019, 30(6):1385
LI H, BAO YR, WANG S, et al. Fingerprint analysis of Radix Rubiae based on multivariate statistical analysis[J]. Lishizhen Med Mater Med Res, 2019, 30(6):1385
[11] 杜义龙, 李艳荣, 洪霞, 等. 基于指纹图谱和多组分含量分析的半枝莲药材质量评价研究[J]. 中草药, 2023, 54(3):1509
DU YL, LI YR, HONG X, et al. Quality evaluation of Scutellaria barbata D. Don based on combination of chromatographic fingerprints and multi-component content[J]. Chin Tradit Herb Drug, 2023, 54(3):1509
[12] 李海峰, 肖凌云, 张菊, 等. 茜草化学成分及其药理作用研究进展[J]. 中药材, 2016, 39(6):1433
LI HF, XIAO LY, ZHANG J, et al. Research progress on chemical constituents and pharmacological effects of Rubiae Radix et Rhizoma[J]. J Chin Med Mater, 2016, 39(6):1433
[13] 刘路, 王佳佳, 崔雪靖, 等. 茜草属植物化学成分研究进展[J]. 中国实验方剂学杂志, 2015, 21(17):225
LIU L, WANG JJ, CUI XJ, et al. Advance on chemical components of Rubia Genus[J]. Chin J Exp Tradit Med Form, 2015, 21(17):225
[14] 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):4
WANG GL, TIAN JG, CHEN DC. Studies on chemical constituents of Rubiae Radix et Rhizoma and R·tinctorum Ⅱ. Determination of alizarin and lucidin by RP-HPLC[J]. Chin J Pharm Anal,1997,17(4):4
[15] 颜洁芳. 茜草的市场调查及质量评价研究[D]. 北京:北京中医药大学, 2021
YAN JF. Study on Market Investigation and Quality Evaluation of Rubiae Radix et Rhizoma[D]. Beijing: Beijing University of Chinese Medicine, 2021
[16] 卫若楠, 吉红玉, 于同月, 等. 茜草的临床应用及其用量探究[J]. 吉林中医药, 2020,40(11):1513
WEI RN, JI HY, YU TY, et al. Study on the clinical application and dosage of Rubiae Radix et Rhizoma[J]. Jilin Tradit Chin Med, 2020, 40(11):1513
[17] 浦益琼, 丁越, 王冰, 等. 液相色谱-质谱法测定茜草中芦西丁含量[J]. 中国中医药信息杂志, 2016, 23(12):96
PU YQ, DING Y, WANG B, et al. Content determination of Lucidin in Rubiae Radix et Rhizoma by LC-MS[J]. Chin J Inf Tradit Chin Med, 2016, 23(12):96
