{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,26]],"date-time":"2026-10-26T20:11:09Z","timestamp":1793045469389,"version":"4.5.0"},"reference-count":34,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T00:00:00Z","timestamp":1777420800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Pharmacol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Astragali radix (AR) is a well-known traditional Chinese medicine (TCM). AR processed with heated honey (HAR) can enhance the effect in treating spleen deficiency. However, some people have used heated sugar as a substitute for heated honey in processing AR. The mechanism underlying the effect of HAR in ameliorating spleen deficiency has not been fully elucidated so far. Therefore, this study aimed to determine whether AR has been processed with heated honey or heated sugar. Subsequently, we compared the differential effects of AR and HAR on the composition of intestinal microbiota, short-chain fatty acid (SCFA) metabolism, and immune responses in spleen-deficient rats.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Rapid evaporative ionization mass spectrometry (REIMS) was employed to detect AR processed with different types of heated honey, different types of heated sugar, and heated honey mixed with heated soft sugar in various proportions. 16S rRNA high-throughput sequencing was conducted to analyze the effects of AR and HAR on intestinal flora diversity in spleen-deficient rats. The regulatory effects of AR and HAR on SCFA metabolism in spleen-deficient rats were measured using headspace gas chromatography-mass spectrometry (HS-GC-MS). The effects of AR and HAR on specific and non-specific immunoreactivity in spleen-deficient rats were evaluated using enzyme-linked immunosorbent assay (ELISA). Furthermore, correlations were detected between different intestinal flora, short-chain fatty acids, and immune response factors.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>We successfully identified AR processed with heated potpourri honey, linden honey, and locust flower honey as genuine heated honey processed products. Additionally, we achieved accurate identification of AR processed with heated soft sugar or granulated white sugar and determined the proportion of adulterated sugar in samples. Both AR and HAR increased the abundance of beneficial gut bacteria, reduced the abundance of pathogenic bacteria, upregulated the synthesis of propionic acid and butyric acid, and modulated the metabolism of isovaleric acid. Notably, the positive effect of HAR on propionic acid expressinon was stronger than that of AR. Meanwhile, both AR and HAR significantly modulated the expression levels of immune factors in serum and colon samples, with HAR exhibiting superior regulatory effects compared to AR.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      The procedure developed in this study can help rapidly and accurately determine whether sugar has been added during the heated honey processing of AR. The enhanced effect of HAR on spleen deficiency may be associated with its regulatory effect on the abundance of\n                      <jats:italic>Lactobacillus<\/jats:italic>\n                      and\n                      <jats:italic>Turicibacter<\/jats:italic>\n                      genera. Through this effect, it promotes the synthesis of propionic acid, upregulates anti-inflammatory factors, downregulates pro-inflammatory factors, and enhances the immunomodulatory effect.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fphar.2026.1711584","type":"journal-article","created":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T05:54:46Z","timestamp":1777442086000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["A novel method for real-time adulteration identification of heated-honey processed Astragali radix using REIMS-iKnife and its regulatory effects on gut microbiota, SCFAs, and immune function in spleen-deficient rats"],"prefix":"10.3389","volume":"17","author":[{"given":"Pengpeng","family":"Liu","sequence":"first","affiliation":[{"name":"Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications, Liaoning University of Traditional Chinese Medicine","place":["Shenyang, Liaoning, China"]},{"name":"School of Pharmacy, Liaoning University of Traditional Chinese Medicine","place":["Dalian, Liaoning, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yisheng","family":"Xu","sequence":"additional","affiliation":[{"name":"Waters Technology (Beijing) Co., Ltd.","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haoran","family":"Yu","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Liaoning University of Traditional Chinese Medicine","place":["Dalian, Liaoning, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tianzhu","family":"Jia","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Liaoning University of Traditional Chinese Medicine","place":["Dalian, Liaoning, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ji","family":"Shi","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Liaoning University of Traditional Chinese Medicine","place":["Dalian, Liaoning, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingyuan","family":"Lu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pengfei","family":"Tu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qian","family":"Cai","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Liaoning University of Traditional Chinese Medicine","place":["Dalian, Liaoning, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,4,29]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1038\/s41538-024-00333-3","article-title":"Identification strategy of wild and cultivated astragali radix based on REIMS combined with two-dimensional LC-MS","volume":"8","author":"Chen","year":"2024","journal-title":"Npj Sci. Food."},{"key":"B2","first-page":"315","article-title":"Pharmacopoeia of the people\u2019s Republic of China","volume-title":"Astragali radix","year":"2020"},{"key":"B3","doi-asserted-by":"publisher","first-page":"650","DOI":"10.1093\/jaoacint\/qsaa168","article-title":"Screening of 154 veterinary drug residues in foods of animal origin using LC\u2013MS\/MS: first action 2020.04","volume":"104","author":"Desmarchelier","year":"2021","journal-title":"J. AOAC Int."},{"key":"B4","doi-asserted-by":"publisher","first-page":"2177","DOI":"10.1080\/14786419.2020.1849197","article-title":"GC-MS and LC-MS: an integrated approach towards the phytochemical evaluation of methanolic extract of pigeon pea Cajanus cajan (L.) Millsp leaves","volume":"36","author":"Dinore","year":"2022","journal-title":"Nat. Prod. Res."},{"key":"B5","doi-asserted-by":"publisher","first-page":"447","DOI":"10.13684\/j.cnki.spkj.2010.12.061","article-title":"Detection of honey adulteration by addition of fructose and glucose using near infrared transflectance spectroscopy","volume":"11","author":"Downey","year":"2003","journal-title":"J. Near Infrared Spectrosc."},{"key":"B6","doi-asserted-by":"publisher","first-page":"4266","DOI":"10.1021\/acs.analchem.9b00807","article-title":"Ambient ionization mass spectrometry: recent developments and applications","volume":"91","author":"Feider","year":"2019","journal-title":"Anal. Chem."},{"key":"B7","doi-asserted-by":"publisher","first-page":"128986","DOI":"10.1016\/j.foodchem.2020.128986","article-title":"Simultaneous determination of 79 pesticides in pigeonpea grains using GC\u2013MS\/MS and LC\u2013MS\/MS","volume":"347","author":"Harischandra","year":"2021","journal-title":"Food Chem."},{"key":"B8","first-page":"210","article-title":"Processing of traditional Chinese medicine","volume-title":"Heated-honey processing","author":"Jia","year":"2008"},{"key":"B9","doi-asserted-by":"publisher","first-page":"525","DOI":"10.3390\/metabo12060525","article-title":"An improved method to quantify short-chain fatty acids in biological samples using gas chromatography-mass spectrometry","volume":"12","author":"Kim","year":"2022","journal-title":"Metabolites"},{"key":"B10","doi-asserted-by":"publisher","first-page":"112376","DOI":"10.1016\/j.foodres.2022.112376","article-title":"LC-IMS-HRMS for identification of biomarkers in untargeted metabolomics: the effects of pterostilbene and resveratrol consumption in liver steatosis, animal model","volume":"165","author":"Lacalle-Bergeron","year":"2023","journal-title":"Food Res. Int."},{"key":"B11","doi-asserted-by":"publisher","first-page":"205","DOI":"10.1016\/j.jtcme.2024.03.003","article-title":"Pediatric tuina treatment for spleen deficiency diarrhea regulated through the skin-brain-gut axis and mast cell degranulation","volume":"15","author":"Li","year":"2024","journal-title":"J. Tradit. Compl. Med."},{"key":"B12","doi-asserted-by":"publisher","first-page":"538","DOI":"10.38212\/2224-6614.3420","article-title":"Analysis of highly polar pesticides in foods by LC-MS\/MS","volume":"30","author":"Liao","year":"2022","journal-title":"J. Food Drug Anal."},{"key":"B13","doi-asserted-by":"publisher","first-page":"104353","DOI":"10.1016\/j.ebiom.2022.104353","article-title":"Atypical E3 ligase ZFP91 promotes small-molecule-induced E2F2 transcription factor degradation for cancer therapy","volume":"86","author":"Liu","year":"2022","journal-title":"EBioMedicine"},{"key":"B14","doi-asserted-by":"publisher","first-page":"1727","DOI":"10.1002\/mas.21778","article-title":"Trends in environmental monitoring of high explosives present in soil\/sediment\/groundwater using LC\u2010MS\/MS","volume":"42","author":"Mary Celin","year":"2023","journal-title":"Mass Spectrom. Rev."},{"key":"B15","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1186\/s40168-019-0704-8","article-title":"Macronutrient metabolism by the human gut microbiome: major fermentation by-products and their impact on host health","volume":"7","author":"Oliphant","year":"2019","journal-title":"Microbiome"},{"key":"B16","doi-asserted-by":"publisher","first-page":"114","DOI":"10.16736\/j.cnki.cn41-1434\/ts.2024.14.036","article-title":"Study on the application of acid hydrolysis in the production of sucrose invert syrup","volume":"30","author":"Ou","year":"2024","journal-title":"Mod. Food."},{"key":"B17","doi-asserted-by":"publisher","first-page":"5892","DOI":"10.3390\/molecules26195892","article-title":"LC-MS\/MS determination of 21 non-steroidal anti-inflammatory drugs residues in animal milk and muscles","volume":"26","author":"Pietruk","year":"2021","journal-title":"Molecules"},{"key":"B18","doi-asserted-by":"publisher","first-page":"115351","DOI":"10.1016\/j.jpba.2023.115351","article-title":"Rapid screening of illegal additives in functional food using desorption electrospray ionization mass spectrometry imaging","volume":"229","author":"Qi","year":"2023","journal-title":"J. Pharm. Biomed. Anal."},{"key":"B19","doi-asserted-by":"publisher","first-page":"170","DOI":"10.3390\/metabo12020170","article-title":"A gas chromatography mass spectrometry-based method for the quantification of short chain fatty acids","volume":"12","author":"Rohde","year":"2022","journal-title":"Metabolites"},{"key":"B20","doi-asserted-by":"publisher","first-page":"341","DOI":"10.1038\/nri.2016.42","article-title":"Gut microbiota, metabolites and host immunity","volume":"16","author":"Rooks","year":"2016","journal-title":"Nat. Rev. Immunol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"111307","DOI":"10.1016\/j.foodres.2022.111307","article-title":"Detection of lipidomics characterization of tuna meat during different wet-aging stages using iKnife rapid evaporative ionization mass spectrometry","volume":"156","author":"Shen","year":"2022","journal-title":"Food Res. Int."},{"key":"B22","doi-asserted-by":"publisher","first-page":"14699","DOI":"10.1021\/acs.jafc.1c05143","article-title":"In situ quality assessment of dried sea cucumber (Stichopus japonicus) oxidation characteristics during storage by iKnife rapid evaporative ionization mass spectrometry","volume":"69","author":"Song","year":"2021","journal-title":"J. Agric. Food Chem."},{"key":"B23","doi-asserted-by":"publisher","first-page":"1629","DOI":"10.1016\/j.foodchem.2012.11.068","article-title":"Detection of adulteration in honey samples added various sugar syrups with 13C\/12C isotope ratio analysis method","volume":"138","author":"Tosun","year":"2013","journal-title":"Food Chem."},{"key":"B24","doi-asserted-by":"publisher","first-page":"132920","DOI":"10.1016\/j.foodchem.2022.132920","article-title":"Recent trends in the analysis of honey constituents","volume":"387","author":"Valverde","year":"2022","journal-title":"Food Chem."},{"key":"B25","doi-asserted-by":"publisher","first-page":"476","DOI":"10.1002\/mas.21915","article-title":"A perspective of multi-reflecting TOF MS","volume":"45","author":"Verenchikov","year":"2024","journal-title":"Mass Spectrom. Rev."},{"key":"B26","volume-title":"Recording summary on traditional Chinese medicine processing methods of all ages","author":"Wang","year":"1998"},{"key":"B27","doi-asserted-by":"publisher","first-page":"1057","DOI":"10.1002\/elps.202300007","article-title":"Lipidomic study and diagnosis of hepatocellular carcinoma tumor with rapid evaporative ionization mass spectrometry","volume":"44","author":"Wang","year":"2023","journal-title":"Electrophoresis"},{"key":"B28","doi-asserted-by":"publisher","first-page":"2515690X241313097","DOI":"10.1177\/2515690X241313097","article-title":"Yiqi Jianpi Kangai decoction enhances the chemotherapy effect by inducing apoptosis and regulating treg and Th17 cells in colorectal cancer mice model with spleen Qi deficiency. J. Evid. Based. Integr","volume":"30","author":"Wang","year":"2025","journal-title":"Med"},{"key":"B29","doi-asserted-by":"publisher","first-page":"224","DOI":"10.3390\/vetsci10030224","article-title":"Study on the hepatoprotective effect mechanism of polysaccharides from charred Angelica sinensis on the layer chickens based on the detection of the intestinal floras and short-chain fatty acids of cecal contents and association analysis","volume":"10","author":"Wu","year":"2023","journal-title":"Vet. Sci."},{"key":"B30","doi-asserted-by":"publisher","first-page":"3133","DOI":"10.3390\/molecules29133133","article-title":"Analysis of antioxidant compounds in Vitex negundo leaves using offline 2D-LC-ECD and LC-MS\/MS","volume":"29","author":"Wu","year":"2024","journal-title":"Molecules"},{"key":"B31","doi-asserted-by":"publisher","first-page":"eabo0789","DOI":"10.1126\/sciadv.abo0789","article-title":"Neuroinflammation inhibition by small-molecule targeting USP7 noncatalytic domain for neurodegenerative disease therapy","volume":"8","author":"Zhang","year":"2022","journal-title":"Sci. Adv."},{"key":"B32","doi-asserted-by":"publisher","first-page":"e9640","DOI":"10.1002\/rcm.9640","article-title":"Effects of ginseng on short-chain fatty acids and intestinal microbiota in rats with spleen-qi deficiency based on gas chromatography-mass spectrometry and 16s rRNA technology","volume":"37","author":"Zhang","year":"2023","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"B33","doi-asserted-by":"publisher","first-page":"575","DOI":"10.3389\/fimmu.2020.00575","article-title":"Intestinal flora and disease mutually shape the regional immune system in the intestinal tract","volume":"11","author":"Zhou","year":"2020","journal-title":"Front. Immunol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"3843","DOI":"10.1016\/j.apsb.2022.06.013","article-title":"Schisandrol A protects AGEs-induced neuronal cells death by allosterically targeting ATP6V0d1 subunit of V-ATPase","volume":"12","author":"Zhou","year":"2022","journal-title":"Acta Pharm. Sin. B"}],"container-title":["Frontiers in Pharmacology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2026.1711584\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T05:54:49Z","timestamp":1777442089000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fphar.2026.1711584\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,29]]},"references-count":34,"alternative-id":["10.3389\/fphar.2026.1711584"],"URL":"https:\/\/doi.org\/10.3389\/fphar.2026.1711584","relation":{},"ISSN":["1663-9812"],"issn-type":[{"value":"1663-9812","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,29]]},"article-number":"1711584"}}