The data acquisition instrument system included devices for ultra-performance liquid chromatography (UPLC, Shim-pack UFLC SHIMADZU CBM30A, https://www.shimadzu.com.cn/ (accessed on 23 October 2022)) and tandem mass spectrometry (MS/MS, Applied Biosystems 4500 QTRAP, http://www.appliedbiosystems.com.cn/ (accessed on 23 October 2022)).
Shim pack uflc cbm30a
Manufactured by Shimadzu
Sourced in Japan
The Shim-pack UFLC SHIMADZU CBM30A is a core module for ultra-fast liquid chromatography (UFLC) systems. It serves as the communication bus module, facilitating the coordination and control of other UFLC system components.
Automatically generated - may contain errors
Lab products found in correlation
4500 q trap, by Thermo Fisher Scientific (3 mentions)
6500 q trap, by Thermo Fisher Scientific (2 mentions)
Acquity uplc hss t3 c18, by Waters Corporation (2 mentions)
Applied biosystems 6500 q trap, by Thermo Fisher Scientific (1 mentions)
4500 q trap, by AB Sciex (1 mentions)
Sb c18 column, by Agilent Technologies (1 mentions)
Scaa 104, by ANPEL (1 mentions)
Acquity uplc hss t3 c18 column, by Waters Corporation (1 mentions)
Qtrap 6500, by AB Sciex (1 mentions)
Qtrap, by AB Sciex (1 mentions)
Qtrap 6500, by Thermo Fisher Scientific (1 mentions)
Vacuum freeze dryer, by Ningbo Scientz Biotechnology (1 mentions)
0.22 μm membrane, by ANPEL (1 mentions)
14 protocols using shim pack uflc cbm30a
1
UPLC-MS/MS Analytical Protocol
2
Metabolic Profiling of Plant Branchlets
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The metabolites were extracted and analyzed by Wuhan Metware Co., Ltd. (https://www.metware.cn/ ). The freeze-dried branchlets were ground at 30 Hz in a mixing mill MM400 (Retsch, Germany) for 15 min. The powder (100 mg) was extracted overnight with 1 ml of 70% methanol (v/v) at 4 °C. After centrifugation at 10,000 g for 10 min, each supernatant was absorbed (Cnwbond Carbon-GCB SPE Cartridge, Anpel, China) and passed through a 0.22-µm pore size filter. The extracted samples were collected and subjected to metabolome analysis by an UPLC system (Shim-pack UFLC Shimadzu CBM30A, Shimadzu, Japan) and a tandem mass spectrometry (MS/MS) system (Applied Biosystems 6500 Q TRAP, Thermo Fisher Scientific, USA). The working parameters of the UPLC and MS were the method as described by [33 (link)]. Based on the Metware database, metabolites were identified according to secondary spectrum information, and metabolites were quantified by multi reaction monitoring of triple quadrupole mass spectrometry. Metabolite abundance was quantified according to peak area. The following thresholds were used to determine whether metabolites differed among treatments (D_0h, D_12h, and D_24h): variable importance in projection (VIP) ≥ 1, P-value < 0.05, fold-change ≥ 2, and fold-change ≤ 0.5]
3
Metabolite Profiling of Transgenic Tobacco
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Metabolite profiling was performed using an LC–ESI–MS/MS system (UPLC; Shim-pack UFLC SHIMADZU CBM30A, Shimadzu, Kyoto, Japan; MS/MS, Applied Biosystems 6500 QTRAP, Applied Biosystems, Foster City, CA, United States) (Chen et al., 2013 (link)). The data from six samples (EbMYBP1-OE and WT × three biological replicates) were processed by OPLS-DA and PCA to detect differences in metabolic composition between the EbMYBP1-OE and WT tobacco.
4
UPLC-ESI-MS/MS Analysis of Prepared Samples
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The prepared sample extraction was analyzed employing a UPLC–ESI–MS/MS system, including a UPLC system (Shim-pack UFLC SHIMADZU CBM30A, Shimadzu, Kyoto, Japan) and an ESI–MS/MS system (Applied Biosystems 4500 Q TRAP, AB SCIEX, Foster City, CA, USA). A separation operation was implemented using a 1.8 µm Agilent SB-C18 column (100 mm × 2.1 mm) equilibrated with mobile phases A and B prepared by mixing ultrapure water with 0.1% formic acid and acetonitrile, respectively. Chromatographic separation was completed by way of a gradient elution program. First, mobile phase B rose linearly from 5% to 95% within 9 min. It was maintained at the level of 95% from 9 to 10 min, decreased to 5% from 10 to 11 min, and held steady until 14 min. The temperature of the column oven was 40 °C, the injection volume was 4 µL, and the flow rate was 0.35 mL/min [47 (link)]. The effluent was alternatively connected to an ESI–triple quadrupole-linear ion trap (QTRAP)-MS.
5
Metabolomic Analysis of Plasma in AAV
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We adopted broadly targeted metabolome technology to analyze the metabolomes of plasma samples from AAV patients with renal involvement and healthy controls. The data acquisition instrumentation system mainly consisted of Ultra Performance Liquid Chromatography (UPLC) (Shim-pack UFLC SHIMADZU CBM30A; Shimadzu, Kyoto, Japan) and tandem mass spectrometry (MS/MS) (4500 QTRAP; Applied Biosystems, Foster City, CA, USA). An ACQUITY UPLC HSS T3 column (2.1 mm i.d. × 100 mm, 1.8 µm; Waters) was used in UPLC to analyze the metabolomes of interest. And the quantification of metabolites was carried out using the multiple reaction monitoring mode of triple quadrupole mass spectrometry. The samples were placed in an autosampler maintained at 40 °C, and then 2 µl samples were injected for LC-MS/MS analysis.
6
UHPLC-MS/MS Analytical Method for Compounds
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Chromatographic separation was carried out on Shim-pack UFLC SHIMADZU CBM30A liquid chromatographic system (Shimadzu, Duisburg, Germany) equipped with a Water ACQUITY UPLC HSS T3 C18 column (1.8 μm, 2.1 mm * 100 mm) at 35°C. A flow rate of 0.4 ml/min was chosen to use while 0.04% acetic water (A) and Acetonitrile (B) with 0.04% acetic acid comprised the mobile phase. Gradient elution program was optimized as follows: 0 ~ 11.0 min, 5% → 95% B; 11.0 ~ 12.0 min, 95% → 95% B; 12.0 ~ 12.1 min, 95% → 5% B; 12.1 ~ 15.0 min, and 5% → 5% B. The injection volume was 5 μl. MS was carried out on an Applied Biosystems 4,500 QTRAP system (AB SCIEX Technologies, United States), equipped with electrospray ionization (ESI) and UHPLC system to scan 100 to 1,500 molecular weight parent ions at 550°C. Other MS parameters were set as follows: MS voltage: 5,500 V; curtain gas (CUR): 25 psi; collision-activated dissociation (CAD): high; declustering potential (DP) and collision energy (CE): specific optimization (Chen et al., 2013 (link)).
7
Quantitative Metabolite Profiling of Maize Kernels
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Ears were taken at 7 days, 14 days, 21 days and 28 days after pollination. Three biological replicates were used for each stage. Grains were isolated from the center of the ears and frozen immediately. The freeze-dried samples were crushed using a mixer mill (MM 400, Retsch) with a zirconia bead for 1.5 min at 30 Hz. Then 100 mg powder was mixed with 1.0 ml 70% aqueous methanol containing 0.1 mg/L lidocaine for internal standard, incubated overnight at 4 °C. Centrifuge at 10,000 g for 10 min, the supernatant was filtrated (SCAA-104, 0.22-µm pore size; ANPEL, Shanghai, China, www.anpel.com.cn/ ) before LC–MS/MS analysis. Quality Control (QC) samples were used to detect reproducibility of the experiment. The compounds were analyzed using an LC-ESI-MS/MS system (UPLC, Shim-pack UFLC SHIMADZU CBM30A; MS/MS, Applied Biosystems 6500 QTRAP,). Data filtering, peak detection, alignment, and calculations were performed using Analyst 1.6.1 software.
8
Metabolite Profiling by UHPLC-MS/MS
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Ultra-high Performance Liquid Chromatography (UHPLC) (Shim-pack UFLC SHIMADZU CBM30A; https://www.shimadzu.com.cn/ ) and tandem mass spectrometry (Applied Biosystems 4500 qtrap; http://www.appliedbiosystems.com.cn/ ) were used to collect metabolite data. The UHPLC conditions were as follows: chromatographic column (Waters ACQUITY UPLC HSS T3 C18 1.8 μm, 2.1 mm * 100 mm); mobile phase A (ultra-pure water with 0.04% acetic acid); mobile phase B (acetonitrile with 0.04% acetic acid); elution gradient (the starting concentration of phase B was 5%; phase B was increased linearly to 95% within 10.00 min and maintained at 95% for 1.0 min; phase B decreased to 5% at 11.00–11.10 min and maintained at 5% for 14 min. Flow rate = 0.35 ml/min; column temperature = 40°C; and injection volume = 4 μL). The mass spectrum conditions were set as follows: the temperature of electrospray ionization was set to 550°C, the voltage was set to 5500 V, the curtain gas pressure was fixed at 30 psi, and the parameter of collision-activated dissociation was set to high. In the triple quadrupole (QQQ), each ion pair was scanned and detected according to the optimized declustering potential and collision energy (Chen et al., 2013 (link)).
9
Metabolomic Analysis of Lycium chinensis
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Root metabolites of L. chinensis from NG, LG, and HG treatments were extracted and analyzed as previously described (Fasano et al., 2016 (link)). Freeze-dried root samples of L. chinensis were put into centrifuge tubes, followed by shaking at 30 Hz for 1.5 min using a Mixer Mill (RetschGmbH, Düsseldorf, Germany) and then overnight extraction at 4°C with 70% aqueous methanol. Samples were centrifuged at 10,000 ×g for 10 min and then filtered (SCAA-104, 0.22-μm pore size; Shanghai, China). Last, filtered samples were analyzed with liquid chromatography–tandem mass spectrometry (LC-MS/MS) (HPLC, Shimpack UFLC SHIMADZU CBM30A; Shimadzu Corporation, Tokyo, Japan). Samples were injected into a Waters ACQUITY UPLC HSS T3 C18 column (2.1 mm × 100 mm, 1.8 µm) with a flow rate of 400 µL/min. Metabolites were identified using the public databases LipidMaps (http://www.lipidmaps.org/ ) and Human Metabolome database (http://www.hmdb.ca/ ) (Wishart et al., 2012 (link)). Differential metabolites were those with variable importance in the projection (VIP) > 1 and p < 0.05.
10
Metabolomic Profiling of Cancer Cells
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Metabolomics and sample collection were performed as previously reported. Briefly, cancer cells were collected and transferred to a 1.5 mL tube to centrifuge at 300 g for 5 minutes at 4°C. Cells were then washed with cold PBS. After centrifugation at 300 g for 5 minutes at 4°C, 1 mL of 80% cold methanol was added and vigorously vortexed to ensure the cell pellet was completely disrupted. The samples were placed on dry ice and moved to a −80°C freezer for 10 minutes, followed by vigorous vortex. The samples were again centrifuged at maximum speed for 10 minutes at 4°C. The supernatant was collected in new tubes and normalized by protein concentration. Supernatants, which also were kept in −80°C until measured, were placed in a new 1.5 ml tube. 100μL of supernatant was used for UPLC-MS/MS. Ultraperformance liquid chromatography, UPLC (Shim-pack UFLC SHIMADZU CBM30A, https://www.shimadzu.com/ ), tandem mass spectrometry, MS/MS (QTRAP@6500+, https://sciex.com/ ).
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