Objective To investigate the differences in metabolites between Rehmannia glutinosa and Helianthus tuberosus before and after steaming to provide a theoretical basis for their identification.

Methods A widely targeted metabolomics technology approach based on UPLC-MS/MS was employed to detect their metabolite components. Principal component analysis (PCA), hierarchical cluster analysis (HCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) were combined to analyze the metabolites.

Results A total of 2,474 metabolites belonging to 12 categories were detected, among which phenolic acids, alkaloids, terpenoids, and flavonoids were dominant. After steaming, the content of sugars— such as manninotriose, raffinose, and D-fructose—and phenolic acids including vanillic acid, syringic acid, and citric acid increased significantly in both raw R. glutinosa and raw H. tuberosus. In contrast, the levels of glycosides, including catalpol, echinacoside, and acteoside, were significantly lower in steamed R. glutinosa than in its raw form. The significantly differential metabolites between the steamed R. glutinosa vs. raw R. glutinosa group were predominantly amino acids and iridoid glycosides, among which the contents of components such as acetylcatalpol, L-tryptophan, and L-lysine decreased significantly in the steamed samples. The differential metabolites between the steamed H. tuberosus vs. raw H. tuberosus group were mainly amino acids and phenolic acids, with components such as ornithine, L-tryptophan, and L-lysine being significantly enriched in raw samples. The significantly differential metabolites between the two raw materials, were primarily amino acids and iridoid glycosides, among which acetylcatalpol was enriched in raw R. glutinosa, while 1-O-feruloylquinic acid and 4-O-feruloylquinic acid were significantly enriched in raw H. tuberosus. The significantly differential metabolites between the steamed R. glutinosa vs. steamed H. tuberosus group were mainly phenolic acids, with components such as 4-O-p-coumaroylquinic acid and 3,5-dicaffeoylquinic acid being significantly higher in steamed H. tuberosus than in steamed R. glutinosa. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of differential metabolites revealed that the metabolites were primarily annotated and enriched in pathways such as amino acid biosynthesis, plant secondary metabolite biosynthesis, and terpenoid biosynthesis.

Conclusion This study elucidates the metabolic characteristics of R. glutinosa and H. tuberosus before and after the steaming process, offering insights from a metabolic profiling perspective for distinguishing between steamed R. glutinosa and steamed H. tuberosus.

1. 曾梦楠, 张宇涵, 郭彭莉, 等. 怀中1号地黄中2-phenylethyl-beta-glucopyranoside通过调节PI3K/Akt/mTOR/HIF-1α通路对低氧性肺动脉高压的影响[J]. 中国中药杂志, 2024, 49(14): 3857-3867.ZengMN, ZhangYH, GuPL, et al. Effect of 2-phenylethyl-beta-glucopyranoside isolated from Huaizhong No.1 Rehmannia glutinosa on hypoxic pulmonary hypertension by regulating PI3K/Akt /mTOR/HIF-1α pathway[J]. China Journal of Chinese Materia Medica, 2024, 49(14): 3857-3867. DOI: 10.19540/j.cnki.cjcmm.20240412.701.

2. 陈亚曼, 刘蕊, 王庆婷, 等. 基于LC-MS结合多元统计分析的栽培地黄与野生地黄中代谢产物特征分析[J]. 中国中药杂志, 2025, 50(20): 5706-5716.ChenYM, LiuR, WangQT, et al. Metabolite characteristics analysis of cultivated and wild Rehmannia glutinosa based on LC-MS combined with multivariate statistical analysis[J]. China Journal of Chinese Materia Medica, 2025, 50(20): 5706-5716. DOI: 10.19540/j.cnki.cjcmm.20250604.101.

3. 徐璐, 狄瑞毅, 薛淑娟, 等. 基于“药效—化学成分群”及“质控系数”的地黄质量评价标准研究[J]. 中国中药杂志, 2025, 50(20): 5717-5732.XuL, DiRY, XueSJ, et al. Quality evaluation standards of Rehmanniae Radix based on the pharmacological activity-chemical component group and quality control coefficient[J]. China Journal of Chinese Materia Medica, 2025, 50(20): 5717-5732. DOI: 10.19540/j.cnki.cjcmm.20250616.101.

4. 赵孟良, 任延靖, 刘颖. PEG-6000模拟干旱胁迫下菊芋HD-Zip基因家族的鉴定及表达分析[J]. 江苏农业科学, 2025, 53(15): 37-44.ZhaoML, RenYJ, LiuY. Identification and expression analysis of the HD-Zip gene family in Helianthus tuberosus under drought stress simulated by PEG-6000[J]. Jiangsu Agricultural Sciences, 2025, 53(15): 37-44. DOI: 10.15889/j.issn.1002-1302.2025.15.006.

5. 田敏, 宗爱珍, 刘丽娜. 菊芋菊糖的特性及应用研究进展[J]. 粮食与油脂, 2022, 35(7): 14-16.TianM, ZongAZ, LiuLN. Research progress on characteristics and application of inulin from Jerusalem artichoke[J]. Cereals & Oils, 2022, 35(7): 14-16. DOI: 10.13194/j.issn.1008-9578.2022.07.004.

6. 王莹, 李娟, 孙雪梅. 菊芋HtMYB2基因VIGS体系构建与功能验证[J]. 中国农学通报, 2024, 40(24): 116-121.WangY, LiJ, SunXM. HtMYB2 gene of Helianthus tuberosus L.: VIGS system construction and functional validation[J]. Chinese Agricultural Science Bulletin, 2024, 40(24): 116-121. DOI: casb2023-0749.

7. QamarTR, SyedF, NasirM, et al. Novel combination of prebiotics galacto-oligosaccharides and inulin-inhibited aberrant crypt foci formation and biomarkers of colon cancer in wistar rats[J]. Nutrients, 2016, 8(8): 465. DOI: 10.3390/nu8080465.

8. 汪悦, 薛夫光, 蒋林树, 等. 菊芋饲料的营养价值、生物活性及其对动物生理功能的调控作用[J]. 动物营养学报, 2020, 32(2): 497-507.WangY, XueFG, JiangLS, et al. Nutritional value and biological activity of Jerusalem artichoke feed and its regulation effects on animal physiological function[J]. Chinese Journal of Animal Nutrition, 2020, 32(2): 497-507. DOI: 10.3969/j.issn.1006-267x.2020.02.003.

9. 张紫怡, 张来宾, 吕洁丽. 地黄的主要化学成分及药理作用研究进展[J]. 新乡医学院学报, 2024, 41(10): 979-986.ZhangZY, ZhangLB, LuJL. Research progress on the main chemical constituents and pharmacological actions of Rehmannia glutinosa[J]. Journal of Xinxiang Medical University, 2024, 41(10): 979-986. DOI: 10.7683/xxyxyxb.2024.10.015.

10. SawadaY, AkiyamaK, SakataA, et al. Widely targeted metabolomics based on large-scale MS/MS data for elucidating metabolite accumulation patterns in plants[J]. Plant Cell Physiol, 2009, 50(1): 37-47. DOI: 10.1093/pcp/pcn183.

11. WuCS, GaoQH, KjelgrenRK, et al. Yields, phenolic profiles and antioxidant activities of Ziziphus jujube Mill. in response to different fertilization treatments[J]. Molecules, 2013, 18(10): 12029-12040. DOI: 10.3390/molecules181012029.

12. 中国药典 2025年版. 四部[S]. 2025: 32.

13. 张晓亚, 徐金娣, 许军, 等. 整合糖组与代谢组学方法比较蒸制和酒炖熟地黄化学成分[J]. 中草药, 2021, 52(6): 1591-1600.ZhangXY, XuJD, XuJ, et al. Integrating glycomics and metabolomics to compare chemical profiles of steaming and wine-stewing processed Rehmanniae Radix Praeparata[J]. Chinese Traditional and Herbal Drugs, 2021, 52(6): 1591-1600. DOI: 10.7501/j.issn.0253-2670.2021.06.006.

14. ZhouL, XuJD, ZhouSS, et al. Integrating targeted glycomics and untargeted metabolomics to investigate the processing chemistry of herbal medicines, a case study on Rehmanniae Radix[J]. J Chromatogr A, 2016, 1472: 74-87. DOI: 10.1016/j.chroma.2016.10.043.

15. 杨思雨, 池玉洁, 张海琼, 等. 基于广泛靶向代谢组学分析不同基原黄连根茎次生代谢产物差异[J]. 中国中药杂志, 2024, 49(20): 5441-5450.YangSY, ChiYJ, ZhangHQ, et al. Widely targeted metabolomics reveals differences of secondary metabolites in rhizomes of different Coptis species[J]. China Journal of Chinese Materia Medica, 2024, 49(20): 5441-5450. DOI: 10.19540/j.cnki.cjcmm.20240714.102.

16. ZhengPF, ZhangM, FangX, et al. Analysis of the fruit quality of pear (Pyrus spp.) using widely targeted metabolomics[J]. Foods, 2022, 11(10): 1440. DOI: 10.3390/foods11101440.

17. 冯卫生. 地黄化学成分及药理作用研究进展[J]. 中医学报, 2025, 40(329): 2035-2055.FengWS. Research progress on chemical composition and pharmacological effects of Dihuang (Rehmannia glutinosa Libosch.)[J]. Acta Chinese Medicine, 2025, 40(329): 2035-2055. DOI: 10.16368/j.issn.1674-8999.2025.10.322.

18. 周国洪, 赵珍东, 汪小根, 等. 基于植物代谢组学的地黄蒸制前后化学成分变化研究[J]. 海峡药学, 2020, 32(7): 34-37.ZhouGH, ZhaoZD, WangXG, et al. Metabolomics study of changes of Rehmannia glutinosa before and after steaming[J]. Strait Pharmaceutical Journal, 2020, 32(7): 34-37. DOI: 10.13194/j.issn.1673-842X.2020.07.012.

19. 管彦辉, 梁正维, 施扬, 等. 滇黄精炮制前后化学成分变化研究[J]. 中草药, 2024, 55(6): 3647-3658.GuanYH, LiangZW, ShiY, et al. Differences in chemical constituents of Polygonatum kingianum before and after processing[J]. Chinese Traditional and Herbal Drugs, 2024, 55(6): 3647-3658. DOI: 10.7501/j.issn.0253-2670.2024.11.008.

20. ZhouL, XuJD, ZhouSS, et al. Chemomics-based marker compounds mining and mimetic processing for exploring chemical mechanisms in traditional processing of herbal medicines, a continuous study on Rehmanniae Radix[J]. J Chromatogr A, 2017, 1530: 232-240. DOI: 10.1016/j.chroma.2017.11.036.

21. 孟祥龙, 王勃, 胡聪, 等. 基于HPLC-MS及网络药理学探讨熟地黄炮制过程与美拉德反应的相关性[J]. 中药材, 2020, 43(1): 61-70.MengXL, WangB, HuC, et al. Study on the correlation between the processing of Rehmanniae Radix Praeporrata and the Maillard reaction based on HPLC-MS and network pharmacology[J]. Journal of Chinese Medicinal Materials, 2020, 43(1): 61-70. DOI: 10.13863/j.issn1001-4454.2020.01.013.

22. 杨庆丽, 张旭, 魏连会, 等. 黑菊芋加工过程中成分变化研究[J]. 食品工业科技, 2018, 39(2): 70-74.YangQL, ZhangX, WeiLH, et al. Changes of main components in the production process of black Jerusalem artichoke[J]. Science and Technology of Food Industry, 2018, 39(2): 70-74. DOI: 10.13386/j.issn1002-0306.2018.02.014.

23. 张志毅. 菊芋中咖啡酰奎宁酸的分离与抗氧化作用研究[D]. 济南: 山东农业大学, 2024.

24. WojdyloA, FigielA, OszmiańskiJ. Effect of drying methods with the application of vacuum microwaves on the bioactive compounds, color, and antioxidant activity of strawberry fruits[J]. J Agric Food Chem, 2009, 57(4): 1337-1343. DOI: 10.1021/jf802507j.