MetaboLights

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ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14146/m_MTBLS14146_LC-MS_alternating_normal-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14146/a_MTBLS14146_LC-MS_alternating_normal-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14146/s_MTBLS14146.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14146/i_Investigation.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14146Based on the self-built targeted standard product database MWDB (metware database), qualitative analysis is conducted according to the retention time RT (Retention time) of the detected substances, the information of daughter and parent ions pairs, and the secondary spectrum data. Metabolite quantification is accomplished by using the multiple reaction monitoring mode (MRM, as shown in the following figure) of the triple quadrupole mass spectrometer for analysis. In the MRM mode, the quadrupole first screens the precursor ions (parent ions) of the target substances, eliminating the ions corresponding to other molecules with different molecular weights to preliminarily eliminate interference; the precursor ions are then induced to ionize in the collision chamber and break down to form many fragment ions. The fragment ions are further filtered and selected by the triple quadrupole to obtain the desired characteristic fragment ion, eliminating the interference of non-target ions and making the quantification more accurate and the repeatability better. After obtaining the metabolite spectral analysis data of different samples, the peak areas of all the mass spectral peaks are integrated, and the peak integration correction is performed for the mass spectral peaks of the same metabolite in different samples. MetaboLightsPublicLiquid Chromatography MS - alternating - normal-phaseThe sample was analyzed by ultra-performance liquid chromatography (Shim-pack UFLC, Shimadzu, CBM30A) and tandem mass spectrometry (QTRAP® 6500+, SCIEX, Framingham, MA, USA). The Column is Waters ACQUITY UPLC HSS T3 C18 1.8 µm.Oregano essential oil improves growth performance and health of steers by maintaining gut-liver axis homeostasis through the ileal microbiota-mediated bile acid pathway.Yongliang HuangCollege of Animal Science and Technology, Gansu Agricultural Universityliverileac contentmass spectrometry assayFollowing thawing, 50 mg of ileal contents and liver tissue were weighed into a 2 mL centrifuge tube, and 500 µL of 70% methanol internal standard extract was added at 4 °C. The mixture was agitated for 3 min and maintained at −20 °C for 30 min. Next, the mixture was centrifuged at 12,000 rpm for 10 min at 4 °C. Subsequently, 250 μL of the supernatant was centrifuged at 12,000 rpm for 5 min at 4 °C. Following this, 150 μL of the supernatant was transferred into the corresponding injection vial for analysis.Bos taurushttps://www.ebi.ac.uk/metabolights/MTBLS14146Yongliang Huang. Gansu Agricultural University. Yingmen Village, Anning District, Lanzhou City, Gansu Province, China. huangyl_gau@163.com.The temperature of the electrospray ionization (ESI) source is 500 °C, the mass spectrometry voltage is 5500 V (positive), -4500 V (negative), the ion source gas I (GS I) is 55 psi, gas II (GS II) is 60 psi, the curtain gas (CUR) is 25 psi, and the collision-activated dissociation (CAD) parameters are set to high. In the triple quadrupole (Qtrap), each ion pair is scanned and detected based on the optimized declustering potential (DP) and collision energy (CE) for each ion pair.Oregano essential oilhuangyl_gau@163.comThe mixed contents of the mid-ileum were collected into four 5-mL sterile tubes and stored at −80 °C for subsequent estimation of metabolomic analyses. A mid-ileum tissue specimen was immediately placed into two 5 mL sterile tubes and stored at −80 ºC for subsequent metabolomic analyses.MetabolomicsMetabolomicsmetabolite profiling assayAB SCIEX API 2000Holstein steersLiverBos taurusileac contentOregano essential oilIleal microorganisms and metabolitesAgilent 1100 Series HPLC SystemFollowing thawing, 50 mg of ileal contents and liver tissue were weighed into a 2 mL centrifuge tube, and 500 µL of 70% methanol internal standard extract was added at 4 °C. The mixture was agitated for 3 min and maintained at −20 °C for 30 min. Next, the mixture was centrifuged at 12,000 rpm for 10 min at 4 °C. Subsequently, 250 μL of the supernatant was centrifuged at 12,000 rpm for 5 min at 4 °C. Following this, 150 μL of the supernatant was transferred into the corresponding injection vial for analysis.Metabolomicsmetabolite profiling assayLiverHolstein steersAB SCIEX API 2000Bos taurusileac contentOregano essential oilIleal microorganisms and metabolitesAgilent 1100 Series HPLC SystemFollowing thawing, 50 mg of ileal contents and liver tissue were weighed into a 2 mL centrifuge tube, and 500 µL of 70% methanol internal standard extract was added at 4 °C. The mixture was agitated for 3 min and maintained at −20 °C for 30 min. Next, the mixture was centrifuged at 12,000 rpm for 10 min at 4 °C. Subsequently, 250 μL of the supernatant was centrifuged at 12,000 rpm for 5 min at 4 °C. Following this, 150 μL of the supernatant was transferred into the corresponding injection vial for analysis.1) Mobile phase: Phase A is ultra-pure water (containing 0.1% formic acid), and Phase B is acetonitrile (containing 0.1% formic acid); 2) Elution gradient: 0 min: water/acetone (95:5 V/V), 11.0 min: 10:90 V/V, 12.0 min: 10:90 V/V, 12.1 min: 95:5 V/V, 14.0 min: 95:5 V/V; 3) Flow rate: 0.4 ml/min; Column temperature: 40 °C; Injection volume: 2 μl.falseOregano essential oil improves growth performance and health of steers by maintaining gut-liver axis homeostasis through the ileal microbiota-mediated bile acid pathwayThe high-concentrate diet presents significant challenges regarding intestinal nutrient absorption and hepatic energy metabolism in beef cattle. Therefore, it is crucial to develop novel intervention strategies to solve it. This study examined the effects of oregano essential oil (OEO) on the growth performance, ileac microorganisms and metabolites, and liver health in steers. Eighteen steers were randomly allocated to two groups (n = 9) and received either a basal diet (CON) or the same diet supplemented with 20 g/(d·head) OEO for 270 days. 2026-03-262026-03-26MTBLS14146