Smilacis Glabrae Rhizoma is a traditional Chinese medicine commonly used for hyperuricemia. Engeletin, one of its major flavonoids, exhibits various pharmacological activities, but its in vivo uric acid-lowering activity and metabolic processes remain unclear. This study aims to elucidate the in vivo existence forms of engeletin and the pharmacological basis underlying its uric acid-lowering effects. The in vivo metabolites of engeletin were identified by using UHPLC-Q-TOF-MS. The xanthine oxidase inhibitory activity was investigated using in vitro enzymatic assays. The in vivo uric acid-lowering effect was evaluated in hyperuricemic mice. A total of 11, 34, 7, 6, and 5 compounds were detected in urine, feces, serum, liver, and kidney samples, respectively. After removing duplicates, 52 compounds were preliminarily identified as in vivo existence forms of engeletin. The main metabolic reaction types were glucuronidation, sulfation, and hydrolysis. Engeletin exhibited no xanthine oxidase inhibitory activity in vitro but possessed uric acid-lowering activity in vivo. Neoisoastilbin and naringenin were metabolites with both xanthine oxidase inhibitory activity and uric acid-lowering activity. The in vivo uric acid-lowering activity of engeletin may be attributable to its two metabolites rather than itself. This study elucidated the pharmacological basis of engeletin and laid the foundation for developing potential therapeutics for hyperuricemia.

为阐明藏药鸡蛋参[植物松叶鸡蛋参（Codonopsis convolvulacea var.pinifolia）的根]的化学成分，该文采用超高效液相色谱-四级杆飞行时间质谱联用（UHPLC-Q-TOF-MS/MS）技术对其化学成分进行定性分析。采用Waters ACQUITY UPLC BEH C 18 （1.7μm, 2.1 mm×150 mm）色谱柱，以0.1 mmol·L -1 乙酸铵水溶液（A）-乙腈（B）为流动相，流速0.3 mL·min -1 ,进样体积3μL,柱温40℃。质谱使用ESI离子源，采用负离子检测模式，扫描范围为m/z 100～1 800 （MS）、m/z 50～1 800 （MS/MS）。根据化合物一级质谱中的准分子离子推测其分子式，根据化合物二级质谱中的特征碎片离子推测其可能的结构片段及分子结构，再结合对照品对比和文献检索，进行结构鉴定。结果表明：（1）首次从松叶鸡蛋参中鉴定出56种化合物，其中含氮类成分6种、苯丙素类成分6种、木脂素类成分22种、黄酮类成分2种、有机酸类成分8种、糖苷或其他类成分12种；采用对照品比对指认了其中11种成分。（2）首次发现木脂素类和苯丙素类化合物为松叶鸡蛋参根的主要成分，并推导出其裂解途径。该研究结果表明，应用UHPLC-Q-TOF-MS/MS技术能够快速高效地初步阐明松叶鸡蛋参根的化学成分，为藏药鸡蛋参的质量标准、体内过程及药效物质等研究提供了化学基础。

The study aims to elucidate the existence forms(original constituents and metabolites) of Notoginseng Radix et Rhizoma in rats and reveal its metabolic pathways. After Notoginseng Radix et Rhizoma was administered orally once a day for seven consecutive days to rats, all urine and feces samples were collected for seven days, while the blood samples were obtained 6 h after the last administration. Using the ultra high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(UHPLC-Q-TOF-MS/MS) technique, this study identified 6, 73, and 156 existence forms of Notoginseng Radix et Rhizoma in the rat plasma, urine, and feces samples, respectively. Among them, 101 compounds were identified as new existence forms, and 13 original constituents were identified by comparing with reference compounds. The metabolic reactions of constituents from Notoginseng Radix et Rhizoma were mainly deglycosylation, dehydration, hydroxylation, hydrogenation, dehydrogenation, acetylation, and amino acid conjugation. Furthermore, the possible in vivo metabolic pathways of protopanaxatriol(PPT) in rats were proposed. Through comprehensive analysis of the liquid chromatography-mass spectrometry(LC-MS) data, isomeric compounds were discriminated, and the planar chemical structures of 32 metabolites were clearly identified. According to the literature, 48 original constituents possess antitumor and cardiovascular protective bioactivities. Additionally, 32 metabolites were predicted to have similar bioactivities by SuperPred. This research lays the foundation for further exploring the in vivo effective forms of Notoginseng Radix et Rhizoma.

Background: Inhibition of intestinal α-glucosidase is a key strategy for controlling postprandial hyperglycemia in diabetes. Astragali Radix (AR), a traditional medicinal and dietary herb widely consumed in China, is rich in flavonoids that are believed to exhibit hypoglycemic properties. Methods: A total of 29 AR-related flavonoids, including both original constituents and metabolites, were screened for α-glucosidase inhibitory activity using in vitro enzymatic assays. Mechanistic investigations were conducted through enzyme kinetics, circular dichroism (CD) spectroscopy, surface plasmon resonance (SPR), and molecular docking. The in vivo hypoglycemic effects were assessed using a postprandial hyperglycemic mouse model. Additionally, potential mixture effects of flavonoid combinations were evaluated. Results: Of the 29 flavonoids, 16 demonstrated significant α-glucosidase inhibitory activity, with five (C3, C17, C19, C28, and C29) identified as novel inhibitors. Structure–activity relationship (SAR) analysis revealed that hydroxylation, particularly at the C-3 position, enhanced activity, while glycosylation and methoxylation reduced it. Mechanistic studies demonstrated that these compounds bind to distinct amino acid residues within the active site of α-glucosidase, inducing conformational changes and exerting different types of inhibition, leading to varying inhibitory mechanisms. Additionally, 15 compounds reduced postprandial blood glucose levels, with C3, C16, C17, C19, and C28 confirmed as novel in vivo inhibitors. Notably, two compositions of flavonoids combined at their individually ineffective concentrations exhibited significant inhibitory effects. Conclusions: This study provides a comprehensive evaluation of AR-related flavonoids as α-glucosidase inhibitors and offers valuable insights for the development of highly effective, low-toxicity, flavonoid-based, antidiabetic therapeutics and functional foods.

Identification of constitutive herbs in an herbal product is critical for ensuring its quality and efficacy. However, current identification methods often lack universality, entail long durations, and involve complex procedures. Therefore, there is an urgent need to develop innovative methods for identifying constitutive herbs. This paper aims to propose a more refined, universal, simple, rapid, and eco-friendly method for identifying more constitutive herbs in herbal products, significantly enhancing the precision and efficiency of quality control method of herbal products. Leveraging UHPLC‒Q‒TOF‒MS (ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry) data of herbs and the R language, we devised algorithms to find exclusive ions and diagnostic ions of each herb. An open platform named IdenHerb for identifying constitutive herbs of herbal products was established. Using IdenHerb, exclusive ions of 94 herbs were found, and from these exclusive ions, their diagnostic ions were screened out. These diagnostic ions were successfully applied to identify constitutive herbs of two mixtures of herbal powders and eight Chinese patent medicines. Furthermore, this methodology was evaluated using three commonly encountered adulterants as test cases. After incorporating the LC‒MS data of these adulterants into the comprehensive LC‒MS database of all herbs, this approach demonstrated excellent discriminatory capabilities. Compared to the Chinese Pharmacopoeia (2020 edition), IdenHerb is capable of identifying a greater number of constituent herbs, thereby enhancing the level of quality control for the herbal products. This strategy deserves further research and wide application.

Ethnopharmacological relevance Astragali Radix-Saposhnikoviae Radix (AR-SR) is a well-known and effective herb pair. Although the compatibility of these two herbs has been widely applied in many traditional Chinese medicine formulas, its potential mechanism still needs to be investigated. Aim of study To evaluate the pharmacokinetic profiles of 10 bioactive compounds derived from AR when administrated alone and in combination with SR to rats, aiming to further reveal the impact of SR on AR. Materials and methods Two groups of male Sprague-Dawley rats received oral administration of AR and AR-SR freeze-dried powder solutions, respectively. UHPLC-QTRAP-MS/MS technology was utilized to perform the pharmacokinetic studies of 10 compounds derived from AR in rat plasma samples. Results A reliable UHPLC-QTRAP-MS/MS method was established to simultaneously determine the rat plasma concentrations of eight isoflavonoids, referring to calycosin (CAL), calycosin-7-O-β-D-glucoside (CAL-G), formononetin (FOR), formononetin-7-O-β-D-glucoside (FOR-G), astrapterocarpan (APC), astrapterocarpan-3-O-β-D-glucoside (APC-G), astraisoflavan-7-O-β-D-glucoside (AIF-G) and formononetin-7-O-β-D-glucuronide (FOR-GN), and two saponins, including astragaloside IV (AS IV) and cycloastragenol (CAG), originating from AR. Following the oral administration of AR, seven isoflavonoids were quickly absorbed but exhibited low plasma concentrations under 17.88 ng/mL except FOR-GN. The latter maintained higher plasma concentration level more than 15 ng/mL for at least 10 h. Besides, for the first time, AS IV was observed with an obvious double-peak phenomenon after administering AR extract, whereas the concentration of CAG was lower than LLOQ before 6 h. When AR and SR were administrated together, the double-peak phenomena of CAL, FOR, APC, AIF-G and FOR-GN were enhanced and there was a significant increase in their values of area under the concentration-time curve (AUC) and mean residence time (MRT) (P < 0.05) while the pharmacokinetic profiles of CAL-G, FOR-G, APC-G, AS IV and CAG stayed almost unchanged (P > 0.05). Moreover, the elimination half-time (t1/2) values of CAL, FOR and APC were significantly elevated, and the clearance rate/bioavailability (CLz/F) for CAL and FOR was reduced (P < 0.05). Conclusions SR has the potential to modulate the ADME process of five out of the eight isoflavonoids (CAL, FOR, APC, AIF-G and FOR-GN, except CAL-G, FOR-G and APC-G) originating from AR. This interaction is especially likely to affect the hepatic and intestinal drug disposition of these isoflavonoids, thereby extending the duration of their pharmacological effects, which may subsequently impact the therapeutic efficacy of AR.

Anthracyclines play an irreplaceable role in cancer treatment, although their clinical application is limited due to severe side effects such as arrhythmia, cardiomyopathy, and myocardial infarction. The currently available clinical drugs for treating anthracycline-induced cardiotoxicity (AIC) are limited by numerous drawbacks, including the side effects of the therapeutic agents, single treatment mechanisms, and individual patient variations. Therefore, novel drugs with broader applicability and multitarget synergistic protective effects are, therefore, urgently needed. Ginsenosides, the primary bioactive constituents of plants belonging to the genus Panax (family Araliaceae), exhibit a wide range of pharmacological activities, including anti-inflammatory, antioxidative, and antitumor effects, and have demonstrated cardioprotective properties against AIC. This article examines the mechanisms of AIC and the modulatory effects of ginsenosides on these mechanisms. This review highlights the potential molecular targets and signaling pathways through which ginsenosides exert therapeutic effects on AIC, including the regulation of oxidative-stress-related pathways such as Keap1/Nrf2, MAPK, STAT, PI3K/Akt, and AMPK; the restoration of mitochondrial function; the modulation of autophagy; and the inhibition of pyroptosis, ferroptosis, and apoptosis. Therefore, this review serves as a theoretical basis and provides a research direction for future investigation regarding the prevention and treatment of AIC with ginsenosides, as well as clinical translation studies.

ETHNOPHARMACOLOGICAL RELEVANCE:Pharmacokinetics is one of the most significant properties for a traditional Chinese medicine (TCM). However, the existing methodologies and concepts cannot fully capture the complexity of a TCM and are often limited by the unavailability of commercial reference compounds. Therefore, it is needed to propose a new strategy for the pharmacokinetics of TCMs that aligns with the holistic perspective. AIM OF STUDY:We aim to elucidate the comprehensive pharmacokinetic profile of TCMs by establishing the strategy of phenomenological pharmacokinetics (PhenoPK). MATERIALS AND METHODS:This strategy encompasses identifying TCM existence forms (original constituents and metabolites) in rat plasma, cluster analysis for pharmacokinetic behaviors of each compound, calculation of PhenoPK parameters for each compound, and acquisition of PhenoPK parameters for TCMs. The Paeoniae Radix Rubra (PRR) and Astragali Radix-Saposhnikoviae Radix (AR-SR) herb pair were selected as case studies in this proof-of-concept investigation. RESULTS:105 PRR and 127 AR existence forms were identified in rat plasma by UHPLC-Q-TOF-MS/MS. Based on the peak areas detected via mass spectrometry rather than actual concentrations, the pharmacokinetic characteristics of these compounds were recorded using a clustered heatmap. The PhenoPK pharmacokinetics of PRR and AR-SR were evaluated using two strategies with 11 PhenoPK parameters, revealing that PRR exhibited rapid absorption, long retention, and slow elimination, while the pharmacokinetic double-peak phenomenon, peak time, and residence time of AR were found to be enhanced by SR. CONCLUSION:This PhenoPK strategy is a simple and novel pharmacokinetic methodology that identifies the holistic pharmacokinetic characteristics of a TCM and offers distinct advantages across several TCM research fields.

To clarify the chemical constituents of the traditional Chinese medicine Aloe made from Aloe barbadensis,i.e.,the concentrated dry matter of the juice of the leaves of A.barbadensis,a systematic qualitative analysis of them was conducted using the technique of HPLC-DAD-ESI-IT-TOF-MSn in conjunction with the comparison of reference compounds and literature search.The gradient elution was performed with water(A)-acetonitrile(B)as mobile phase at a flow rate of 1.0 mL·min-1.The liquid chromatography-mass spectrometry data were acquired under alternate negative ion and positive ion detection mode using an ESI ion source.The structure elucidation of the chemical constituents was mainly based on negative ion mass spectrometry data.The results were as follows:(1)For the first time,the fragmentation pathways of anthraquinones(aloe-emodin,physcion,and emodin-8-O-β-D-glucoside),anthrones(aloin A,aloinoside A),chromones(aloeresin D,7-O-methylaloeresin A,altechromone A,aloesin,aloeresin G,and aloeresin C),and α-pyranones(aloenin A,aloenin B)in Aloe made from A.barbadensis were clarified.The fragmentation pathway of anthraquinones was dominated by loss of CO2 and CO,and that of anthrones was dominated by cleavage of hexosides and loss of CO.The fragmentation pathways of chromones was dominated by cleavage of hexosides and hydrolysis of the ester group,and that of α-pyranones was dominated by cleavage of hexosides and loss of CO2 and H2O.(2)A total of 168 chemical constituents of Aloe made from A.barbadensis were detected,and 78 of them were identified on the basis of reference compound comparison,literature retrieval,and chemical database(such as SciFinder)searching.The 78 compounds included 3 anthraquinones,29 anthrones,35 chromones,7 α-pyranones and 4 other constituents.Twenty-three of 78 compounds were discovered in the leaves of A.barbadensis for the first time.Fourteen of 23 were newly discovered compounds,including aloinoside D,isoeleutherin,and ethylidene-aloenin,possessed antibacterial,anti-inflammatory,or free radical scavenging activities.The results of this study further enrich the information on the chemical constituents of the traditional Chinese medicine Aloe made from A.barbadensis,and lay a foundation for the study of the therapeutic material basis and quality control methods of Aloe.

Notoginsenoside R1 (NG-R1) is a primary active constituent in Panax notoginseng, a medicinal and edible plant. It is a saponin with protopanaxatriol (PPT) as its aglycone. UHPLC-ESI-Q-TOF-MS/MS was used to clarify the existing forms of NG-R1 and their distributions in rats. The nomenclature of the ESI MS fragmentation pathway and ions of PPT was proposed for the first time. Totally, 105 metabolites with 89 new metabolites were identified. In terms of their LC-MS data, 7 were accurately identified by comparison with reference compounds, and 41 were clearly identified. Polyhydroxylation, pentosylation, acetylation, glucuronidation, and amino acid conjugation are new metabolic reactions of NG-R1. In total, 69, 48, 47, 43, 24, 15, and 7 metabolites were detected in the large intestine, stomach, small intestine, liver, lungs, kidneys, and heart, respectively. Then, 48 metabolites were predicted to be effective by PharmMapper, and their mechanisms of action on three diseases were predicted by network pharmacology. Finally, the antitumor effects on cell proliferation and the anti-inflammatory effects of the eight compounds were verified by cellular experiments. These results help further elucidate the in vivo existing forms of dammarane-type triterpenoids and form the basis for discovering their effective forms in the future.

Germacrone and curdione are germacrane-type sesquiterpenoids that are widely distributed and have extensive pharmacological activities; they are the main constituents of 'Xing-Nao-Jing Injection' (XNJ). Studies on the metabolic features of germacrane-type sesquiterpenoids are limited. In this study, the metabolites of germacrone and curdione were characterized by UHPLC-Q-Exactive Oribitrap mass spectrometry after they were orally administered to rats. In total, 60 and 76 metabolites were found and preliminarily identified in rats administered germacrone and curdione, respectively, among which at least 123 potential new compounds were included. New metabolic reactions of germacrane-type sesquiterpenoids were identified, which included oxidation (+4 O and +5 O), ethylation, methyl-sulfinylation, vitamin C conjugation, and cysteine conjugation reactions. Among the 136 metabolites (including 113 oxidation metabolites, two glucuronidation, two methylation, nine methyl-sulfinylation, three ethylation, six cysteine conjugation, and one Vitamin C conjugation metabolites), 32 metabolites were detected in nine organs, and the stomach, intestine, liver, kidneys, and small intestine were the main organs for the distribution of these metabolites. All 136 metabolites were detected in urine and 64 of them were found in feces. The results of this study not only contribute to research on in vivo processes related to germacrane-type sesquiterpenoids but also provide a strong foundation for a better understanding of in vivo processes and the effective forms of germacrone, curdione, and XNJ.

Drug toxicity is a major concern in medicine, and toxicity is an unresolved obstacle to the efficacy of chemotherapy drugs for cancer. Based on our ‘additive effect’ and ‘toxicity scattering effect’ hypothesis, we propose a unique drug-combination strategy to overcome drug toxicities: combining 8–10 drugs that act on the same target at ineffective doses (much lower than their usual doses) to dramatically reduce the toxicity of each drug without weakening the efficacy of the combination. Using this strategy, we developed an anticancer combination of 8 drugs (topoisomerase II inhibitors); each drug was included at 7.7%–19.5% of its minimum effective dose. This combination prolonged life by 92.8% in ascites tumour-bearing mice and inhibited tumour growth by 54.3% in solid tumour-bearing mice, without causing mortality or detectable liver, cardiac, or renal toxicity. At an average of 1/3 of the combination dose, each drug alone resulted in ineffective tumour inhibition (only 24.6%–34.5% reduction) but caused a 10%–50% mortality. As number of drugs in the combination increased from 1 to 4 to 8, the dose of each drug and the overall toxicity progressively decreased. Combinations of four or six topoisomerase II inhibitors at low concentrations inhibited the proliferation of cancer cell lines and the activity of topoisomerase II, revealing an evident additive effect on the same target. These results demonstrate the significant advantage of this strategy in reducing toxicity. Although this strategy violates drug use principles, it may be a breakthrough in reducing toxicity and improving cancer treatment. This strategy can also be applied to other diseases with high drug toxicity.

This study explored the existence forms(original constituents and metabolites)of Tiantian Capsules,Aloe,and Tiantian Capsules without Aloe in rats for the first time,aiming to clarify the contribution of Aloe to the existence form of Tiantian Capsules.Rats were administrated with corresponding drugs by gavage once a day for seven consecutive days.All urine and feces samples were collected during the seven days of administration,and blood samples were collected 0.5,1,and 1.5 h after the last administration.UHPLC-Q-TOF-MS was employed to detect and identify the original constituents and metabolites in the samples.A total of 34,28,and 2 original constituents and 64,94,and 0 metabolites were identified in the samples of rats administrated with Aloe,Tiantian Capsules,and Tiantian Capsules without Aloe,respectively.The main metabolic reactions were methylation,hydrogenation,hydroxylation,dehydroxylation,glucuronidation,and sulfation.This study clarified for the first time the existence forms and partial metabolic pathways of Aloe,Tiantian Capsules,and Tiantian Capsules without Aloe in rats,laying a foundation for revealing their effective forms.The findings are of great significance to the research on the functioning mechanism and quality control of Aloe and Tiantian Capsules.

INTRODUCTION:Smilacis Glabrae Rhizoma (SGR) is rich in chemical constituents with a variety of pharmacological activities. However, in-depth research has yet to be conducted on the chemical and pharmacodynamic constituents of SGR. MATERIALS AND METHODS:In this study, the chemical constituents of SGR were analyzed using liquid chromatography-mass spectrometry, and the pharmacodynamic compounds responsible for the medicinal effects of SGR were elucidated through a literature review. RESULTS:In total, 20 potentially new compounds, including 16 flavonoids (C19, C20, and C27-C40) and four phenylpropanoids (C107, C112, C113, and C118), together with 161 known ones were identified in the ethanol extract of SGR using liquid chromatography-mass spectrometry, and 25 of them were unequivocally identified by comparison with reference compounds. Moreover, 17 known constituents of them were identified in the plants of genus Smilax for the first time, and 16 were identified in the plant Smilax glabra Roxb. for the first time. Of 161 known compounds, 84 constituents (including isomers) have been reported to have 17 types of pharmacological activities, covering all known pharmacological activities of SGR; among these 84 bioactive constituents, six were found in the plants of genus Smilax for the first time and five were found in S. glabra for the first time, which are new bioactive constituents found in the plants of genus Smilax and the plant S. glabra, respectively. CONCLUSION:The results provide further information on the chemical composition of SGR, laying the foundation for the elucidation of the pharmacodynamic substances of SGR.

Drynariae Rhizoma has been used to treat bone diseases and kidney deficiency in traditional medicine. Recently its aqueous extract was reported to enhance memory function. Although the Japanese standards for non-Pharmacopoeial crude drugs 2022 prescribed Drynaria roosii as the botanical origin, some counterfeits and both raw and stir-fired crude drugs are available in markets. To distinguish Drynariae Rhizoma derived from D. roosii appropriately from others and verify the validity of uses of stir-fried ones, 1 H NMR-based metabolite profiling coupled with HPLC were performed. Raw samples derived from D. roosii contained naringin ( 1 ), neoeriocitrin ( 2 ), 5,7-dihydroxychromone-7- O -neohesperidoside ( 3 ), caffeic acid 4- O -β- d -glucoside ( 4 ), protocatechuic acid ( 5 ), trans- p -coumaric acid 4- O -β- d -glucoside ( 6 ), and kaempferol 3- O -α- l -rhamnoside 7- O -β- d -glucoside ( 8 ). Stir-fried samples were characterized by presence of 5-hydroxymethyl-2-furaldehyde ( 13 ), and were divided into two types; one possessing similar composition to raw samples (Type I) and another without above components except 5 (Type II). Quantitative analyses using qHNMR and HPLC, followed by principal component analysis demonstrated that the raw samples had higher contents of 1 (0.93–9.86 mg/g), 2 (0.74–7.59 mg/g), 3 (0.05–2.48 mg/g), 4 (0.27–2.51 mg/g), 6 (0.14–1.26 mg/g), and 8 (0.04–0.52 mg/g), and Type II had a higher content of 5 (0.84–1.32 mg/g). The counterfeit samples derived from Araiostegia divaricata var. formosana were characterized by higher content of ( −)-epicatechin 3- O -β- d -allopyranoside ( 10 ) (1.44–11.49 mg/g) without 1 and 2 . These results suggested that Drynariae Rhizoma samples derived from other botanical origins and Type II stir-fried samples cannot substitute for D. roosii rhizome.

Supplementary Figures S1-S5. Figure S1. SW shows good anticancer activity among 6 fractions from SGR. A, flow chart for preparation of SW from sarsaparilla. Figure S2. SW inhibits cancer cell growth. A, MTT screening with 11 cell lines to evaluate the growth inhibitory effect of SW. B, colony formation of AGS, HT-29, H1299, T24, PG, HepG2 and PC3 cells treated with indicated concentrations of SW. Figure S3. A, left, pictures of H22 allografts in PBS-, CTX- (80 mg/kg once a week) and SW-treated (72.7 mg once a day) groups. Figure S4. SW-induced oxidative stress is unrelated to H2O2 or NO. A, DCFH-DA labeling of intracellular ROS in SW- or solvent-treated AGS and HT-29 cells. Figure S5. ERK is downstream of GSH/GSSG imbalance upon SW treatment. A, U0126 inhibited SW-induced ERK1/2 phosphorylation. B, the effect of U0126 on SW-induced GSH/GSSG imbalance. Columns, mean; bars, SD.

Supplementary Table S1 and Figure legends. Supplementary Table S1 of primer sequences and Figure legends for Figure S1-S5.

Mogrosides III (1) and IIIE (2) are two important bioactive cucurbitane-type triterpenoid triglycosides in the edible fruits of Siraitia grosvenorii (Swingle), which are isomers and have only a minor difference in their structures. To clarify the effects of structural difference and drug-metabolizing-enzyme induction on their metabolism in vivo, their metabolites in normal rats and drug-metabolizing-enzyme-induced rats were tentatively identified and semiquantified by using the HPLC-DAD-ESI-IT-TOF-MSn technique. Totally, 76, 78, 96, and 121 metabolites of mogrosides were identified in the NIII (normal rats orally administered with mogroside III), NIIIE (normal rats orally administered with mogroside IIIE), EIII (drug-metabolizing-enzyme-induced rats orally administered with mogroside III), and EIIIE (drug-metabolizing-enzyme-induced rats orally administered with mogroside IIIE) groups, respectively. The metabolite differences among these groups indicated that their minor structural differences are responsible for the significant differences in their metabolites, and the induction of drug-metabolizing enzymes significantly increased the number of their metabolites. These findings would improve our understanding of the in vivo processes of mogrosides III and IIIE as well as their interactions with other food bioactive components or drugs.

Introduction: Sanjin tablets (SJT) are a well-known Chinese patent drug that have been used to treat urinary tract infections (UTIs) for the last 40 years. The drug consists of five herbs, but only 32 compounds have been identified, which hinders the clarification of its effective substances and mechanism.Methods: The chemical constituents of SJT and their effective substances and functional mechanism involved in the treatment of UTIs were investigated by using high performance liquid chromatography-electrospray ionization-ion trap-time of flight-mass spectrometry (HPLC-ESI-IT-TOF-MSn), network pharmacology, and molecular docking.Results: A total of 196 compounds of SJT (SJT-MS) were identified, and 44 of them were unequivocally identified by comparison with the reference compounds. Among 196 compounds, 13 were potential new compounds and 183 were known compounds. Among the 183 known compounds, 169 were newly discovered constituents of SJT, and 93 compounds were not reported in the five constituent herbs. Through the network pharmacology method, 119 targets related to UTIs of 183 known compounds were predicted, and 20 core targets were screened out. Based on the "compound-target" relationship analysis, 94 compounds were found to act on the 20 core targets and were therefore regarded as potential effective compounds. According to the literature, 27 of the 183 known compounds were found to possess antimicrobial and anti-inflammatory activities and were verified as effective substances, of which 20 were first discovered in SJT. Twelve of the 27 effective substances overlapped with the 94 potential effective compounds and were determined as key effective substances of SJT. The molecular docking results showed that the 12 key effective substances and 10 selected targets of the core targets have good affinity for each other.Discussion: These results provide a solid foundation for understanding the effective substances and mechanism of SJT.