Biomedical Chromatography

Yuning, Zhu; Runbin, Sun; Xinwen, Zhang; Haoyi, Yang; Yuwen, Zhang; Fei, Fei; Juan, Li

doi: 10.1002/bmc.70195pmid: 40789767

Kaempferol is a natural flavonoid with low bioavailability, but it demonstrates significant anti‐inflammatory properties. In a DSS‐induced colitis model, oral administration of kaempferol effectively alleviated characteristic symptoms of ulcerative colitis (UC) in mice. However, its regulatory effects on metabolism within the circulatory system, colon, and gut microenvironment remain insufficiently explored. Pharmacokinetic properties and metabolomics analysis revealed that the much higher level of kaempferol in the gut contents may contribute to its more pronounced metabolic regulatory effects on gut contents compared to those observed in the serum and colon. In detail, kaempferol significantly reversed 102 metabolites in gut contents, involving metabolic pathways comprising amino acid, bile acid, fatty acid, and nucleotide metabolism. Conversely, kaempferol modulated only 10 metabolites in serum and 17 in colon. The systemic effects of kaempferol mediated via gut‐host crosstalk were evidenced by the regulation of shared metabolic pathways. These included tryptophan metabolism and primary bile acid biosynthesis in both serum and gut contents, as well as linoleic acid metabolism and biosynthesis of unsaturated fatty acids in both colon and gut contents. These insights provide a mechanistic basis for the anti‐colitic effects of kaempferol and identify potential metabolic targets for therapeutic intervention in UC within the intestinal ecosystem.

Mammadova, Gunay; Koseoglu‐Yilmaz, Pelin

doi: 10.1002/bmc.70173pmid: 40693429

In the present study, it was aimed to optimize the extraction of fumaric acid from Fumaria officinalis L. by response surface methodology and its quantification by a newly developed and validated reverse phase high performance liquid chromatography method. Chromatographic separations were performed at 40°C with a C18 column. Aqueous o‐phosphoric acid solution and methanol were used as the mobile phase by gradient elution. The flow rate was set at 1.0 mL/min, and the injection volume was 20 μL. Fumaric acid was detected at 215 nm with a UV detector. The method was linear in the range of 10.00–200.00 μg/mL. The limits of detection and quantification were 3.02 μg/mL and 9.15 μg/mL, respectively. The relative mean errors were ≤ 3.77%, whereas the relative standard deviation values were calculated as ≤ 0.48%. The conditions to obtain an extract with high fumaric acid content were optimized by central composite design, a type of response surface methodology. The independent variables were extraction time, solvent volume, and temperature for the maceration procedure, where fumaric acid concentration was the response. The experimental model was evaluated statistically and found to fit well, considering the lack‐of‐fit p value of 0.211 and coefficient of determination 0.9862 at a confidence level of 95%.

Wei, Shuyun; Zhang, Shuiping; Luo, Jiahui; Yu, Wenxin; Zeng, Xing; Lan, Lunli; Yu, Cuiping; Zeng, Yu; Feng, Yi

doi: 10.1002/bmc.70166pmid: 40702587

Non‐alcoholic fatty liver disease (NAFLD) has become a prominent public health concern, closely linked to metabolic syndromes. Liupao tea (LT), a traditional Chinese dark tea, has demonstrated hepatoprotective effects by regulating metabolism. This study investigated the protective effects of LT on HFD‐induced NAFLD using metabolomics and network pharmacology approaches. We found that LT significantly reduced energy intake in HFD‐fed rats, attenuated abnormal visceral fat accumulation, and prevented hyperlipidemia, abnormal liver lipid deposition, and liver steatosis. Serum untargeted metabolomic analysis identified 46 differential metabolites as potential biomarkers associated with 10 metabolic pathways, including arginine and proline metabolism. Network pharmacology suggested that LT exerts its hepatoprotective effects by regulating arginine metabolism and inflammatory factors; key components, such as caffeine and epigallocatechin gallate, showed direct relevance to NAFLD. Following the intervention, targeted metabolomic analysis revealed a significant change in the levels of 18 relevant amino acids, confirming LT's impact on arginine metabolism. Immunohistochemical results demonstrated reduced expression of inflammatory factors (TNF‐α, IL‐6, IL‐1β) in the liver, suggesting improved liver health. Collectively, these findings indicate that LT mitigates HFD‐induced NAFLD through regulation of amino acid metabolism and reduction of inflammatory factors, thereby alleviating liver injury.

Guoshuang, Peng; Lili, Wan; Xiang, Liu; Bei, Li; Yongbing, Sun

doi: 10.1002/bmc.70183pmid: 40754604

In this study, a novel RP‐HPLC–MS/MS method was developed for the accurate quantification of baloxavir in rat plasma. The analytes were retained and separated on a C18 column, and a triple‐quadrupole mass spectrometer equipped with an electrospray ionization (ESI) source was applied for quantification. Baloxavir could be well retained in the C18 column under the appropriate chromatographic conditions. Using a simple protein precipitation method, the extraction recovery was about 100%, and the ME could be negligible. The method demonstrated excellent linearity (r = 0.9995) across the concentration range of 1.59 ~ 3180.00 ng/mL for baloxavir. The precision and accuracy were within acceptable limits. The lowest limit of quantification (LLOQ) was determined to be 1.59 ng/mL. This simple method was validated and successfully applied to support the pharmacokinetics study after baloxavir was administered to the Sprague–Dawley rats.

Xu, Zhenzhen; Hou, Jingyi; Li, Jing; Bi, Shijie; Wang, Zewen; Liu, Chaoqun; Liu, Yanxia; Yu, Bin; Tian, Jiaye; Yang, Yongjiang; Qiao, Liansheng; Qiao, Yanjiang; Zhang, Yanling

doi:

Gharat, Mithun M.; Roy, Pallavi T.; Gosar, Amit N.; Shaikh, Tabrez A.; Wadhava, Gurumeet C.; Mirgane, Nitin A.

doi: 10.1002/bmc.70189pmid: 40771022

An LC–MS (liquid chromatography–mass spectrometry) methodology is developed for the precise analysis of dichloroacetic acid (DCAA) genotoxic impurity with high sensitivity and selectivity in a Cetirizine dihydrochloride (CTZ) drug substance. In accordance with the “threshold of toxicological concern (TTC),” the carryover of DCAA in a CTZ is done at a limit of 25 ppm in the sample. The present methodology is validated as per “International Council for Harmonization (ICH)” guidelines, and the detection limit and the quantitation limit are 0.4 and 1.2 ppm, respectively. The linearity study was conducted, and the coefficient of regression was found to be 0.9946. The method's accuracy was confirmed by the finding the percentage of recovered spiked DCAA in the drug, which ranged from 93.47% to 99.80%. Results indicated that the methodology was reliable, precise, and reproducible. The method can be extended for the determination of DCAA genotoxic impurity in a CTZ drug substance (API) samples by LC–MS.

Qian, Xue; Liu, Jiaqi; Yang, Muyi; Yue, Jiayin

doi: 10.1002/bmc.70186pmid: 40765222

Gardenia jasminoides Ellis root, a traditional Chinese medicine, exhibits various pharmacological activities, including hepatoprotective and anti‐inflammatory. It is also effective in the treatment of nonalcoholic fatty liver disease. However, its pharmacological substances and mechanisms for alleviating cognitive dysfunction in nonalcoholic fatty liver disease remain unclear. This study explored the pharmacological substances and mechanisms of Gardenia jasminoides Ellis root in improving cognitive impairment in nonalcoholic fatty liver disease using serum medicinal chemistry, network pharmacology, molecular docking, and molecular dynamics simulation. Serum medicinal chemistry analysis was used to identify the blood components of Gardenia jasminoides Ellis root. Network pharmacology analysis further revealed interactions between its active ingredients and targets related to disease. Additionally, molecular docking experiments demonstrated that the active compounds genistein, glaucine, and 3‐(benzyloxy)aniline interact with steroid hormone receptors, including HSP90AA1, AKT1, and EP300. At last, the molecular dynamics simulation of the HSP90AA1–genistein complex with high binding energy was carried out, and the binding of the complex was stable. The above results indicated that the core component genistein and HSP90AA1 protein complex in Gardenia jasminoides Ellis root may indirectly regulate nonalcoholic fatty liver cognitive impairment through the neuroactive ligand–receptor interaction, cAMP signaling pathway, and HIF‐1 signaling pathway.

Zhang, Yu; Shao, Li‐Yang; Li, Yu; Li, Jiayi; Hui, Lingyun; Zhang, Jing

doi: 10.1002/bmc.70176pmid: 40717230

The use of volatile organic compounds (VOCs) as noninvasive disease biomarkers has been explored as the basis for the diagnosis of human diseases. Although VOCs are of interest clinically, limited studies have been conducted because identifying disease‐related VOCs and neglecting irrelevant variance in the data is still a formidable challenge. In our work, the VOCs in the headspace of breast cell medium and human exhaled gas were analyzed by solid‐phase microextraction—gas chromatography—mass spectrometry. The common VOCs, which may be the breast cancer (BC)‐associated VOCs, were identified by statistical analysis, including 1‐butanol, cyclohexanone, nonanal, acetophenone, and benzyl alcohol. Then, after treatment of MCF‐7 cells with letrozole, the levels of four VOCs were reduced. Five VOCs in exhaled breath of BC patients with drug therapy were decreased. It further proves that these VOCs are closely BC‐associated VOCs. The study screened five BC‐associated VOCs from in vitro cell VOCs and human exhaled breath, and provides basic research for VOCs that can be used in the screening of clinical diseases.

Louati, Kaouthar; Kolsi, Fatma; Mellouli, Manel; Louati, Hanen; Kallel, Rim; Zribi, Rania; Borni, Mahdi; Hakim, Leila Sellami; Maalej, Amina; Choura, Sirine; Chamkha, Mohamed; Sayadi, Sami; Khemakhem, Zouheir; Boudawara, Tahya Sellami; Boudawara, Mohamed Zaher; Zribi, Kaouthar; Safta, Fathi

Lee, Seungyeon; Lee, You‐Rim; Mun, Sora; Yun, Yeeun; Kang, Hee‐Gyoo; Lee, Jiyeong

doi: 10.1002/bmc.70197pmid: 40797350

Major depressive disorder (MDD) is a mood disorder that causes serious functional impairment. Existing diagnostic methods rely on subjective assessments because of its complex and heterogeneous pathophysiology; therefore, development of objective biomarkers is urgently needed. To control the high heterogeneity of MDD, a pairwise design in which depressed and remitted states of the same patient were paired to minimize the influence of intrinsic factors was introduced, and serum metabolite changes between states were analyzed using non‐targeted and targeted metabolomics approaches. State‐based biomarkers that could be used for objective diagnosis of MDD were identified, and their clinical applicability was validated in an expanded study group. L‐Glutamine was selected because it showed a tendency to increase during the remitted state. Through multiple reaction monitoring‐based quantitative verification, L‐glutamine levels significantly increased in the remitted state compared with those in the depressed state, regardless of the influence of drug treatment, proving its potential as a diagnostic marker. This study identified differences in serum metabolites in patients with MDD using a metabolomic approach; L‐glutamine could be used as a promising biomarker to distinguish between depressive and remissive states. These results can contribute to the precise diagnosis of MDD and establishment of personalized treatment strategies.