1290 uplc system

Manufactured by Agilent Technologies

Sourced in United States

The 1290 UPLC system is a high-performance liquid chromatography (HPLC) instrument designed for efficient separation and analysis of chemical compounds. It features advanced technology for improved resolution, sensitivity, and speed compared to traditional HPLC systems.

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Agilent 1290 UPLC system (55 citations) 1290 Infinity UPLC system (29 citations) 1290 UPLC (18 citations) Agilent 1290 Infinity UPLC system (15 citations) Agilent 1290 series UPLC system (14 citations) 1290 Infinity II UPLC system (13 citations) UPLC system (8 citations) Agilent 1290 Infinity II UPLC system (7 citations) 1290 Infinity series UPLC System (3 citations) 1290 series UPLC system (3 citations)

43 protocols using 1290 uplc system

UPLC-QTOF-MS/MS Fingerprinting of TCMs

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Ultrahigh performance liquid chromatography coupled with quadrupole time flight mass spectrometry (UPLC–QTOF-MS/MS) analysis was performed on an Agilent 1290 UPLC system coupled to Agilent 6545 Q-TOF instrument (Agilent, Santa Clara, CA, USA). The spectrometer was operated in full-scan TOF–MS at m/z 50–1500 and information-dependent acquisition (IDA) MS/MS modes. The specific conditions were: gas temperature (320 °C), Drying Gas (11 L/min), Nebulizer (35 psi), Shealth Gas Temp (350 °C), Shealth Gas Flow (11 L/min), and Collision Energy (40 eV). Chromatographic separation was achieved using an Agilent ZORBAX RRHD Eclipse Plus C18 (3.0 × 100 mm, 1.8 μm) analytical column. The mobile phase consisted of 0.1% formic acid (v/v) in water and acetonitrile. The temperature (°C), detection wavelength (nm), and velocity of flow (mL/min) were 30, 260 and 0.5 for RG, 40, 215 and 0.3 for GJ, 30, 190–400 and 0.3 for FZ, 25, 260 and 0.3 for DH, and 30, 280 and 0.3 for HQ. For HL, HPLC analysis was obtained for stable baseline and coupled to Agilent ZORBAX Extend-C18 (4.6 mm × 250 mm, 5 µm, 25 °C, sample injection volume 2 µL). The mobile phase consisted of acetonitrile and 0.1% trifluoroacetic acid solution. Each TCM was analyzed by a specific gradient elution procedure.

Li B., Tao X., Sheng L., Li Y., Zheng N, & Li H. (2022). Divergent impacts on the gut microbiome and host metabolism induced by traditional Chinese Medicine with Cold or Hot properties in mice. Chinese Medicine, 17, 144.

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Quantitative Analysis of Endocannabinoids by UPLC-MS/MS

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Anandamide (AEA) and 2-arachidonylglyceriol (2-AG) were quantified using modified ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) by Lam method [35] (link). Octadeuterated endocannabinoids: AEA-d8 and 2-AG-d8 as internal standards were added into the homogenate and all cannabinoids were isolated using a solid phase extraction (SPE). UPLC–MS/MS analysis was performed using an Agilent 1290 UPLC system with a Zorbax Extend C18 column (2.1 mm×150 mm, 1.8 mm, Agilent, Santa Clara, CA, USA) and interfaced with an Agilent 6460 triple quadrupole mass spectrometer with electrospray ionisation source (ESI). The samples were analysed in positive-ion mode using multiple reaction monitoring (MRM). Transition of the precursor to the product ion for AEA was: m/z 348.3→62.1, and for 2-AG: m/z 379.3→287.2. The linear dynamic range was 0.2–200 ng/g for AEA and 0.8–100 μg/g for 2-AG.The LOQ value was 0.2 ng/g for AEA, and 0.8 μg/g for 2-AG, while the LOD values were 0.02 ng/g and 0.08 μg/g, respectively.

Gęgotek A., Nikliński J., Žarković N., Žarković K., Waeg G., Łuczaj W., Charkiewicz R, & Skrzydlewska E. (2016). Lipid mediators involved in the oxidative stress and antioxidant defence of human lung cancer cells. Redox Biology, 9, 210-219.

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Quantification of Endocannabinoids via UPLC-MS/MS

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Anandamide (AEA) and 2-arachidonoylglycerol (2-AG) were quantified using modified ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) by the Lam method [39 (link)]. Octadeuterated endocannabinoids AEA-d₈ and 2-AG-d₈ were added as internal standards to the cell lysates, and all cannabinoids were isolated using solid phase extraction (SPE). UPLC–MS/MS analysis was performed using an Agilent 1290 UPLC system with a Zorbax Extend C18 column (2.1 × 150, 1.8 mm, Agilent, Santa Clara, CA, USA) and interfaced with an Agilent 6460 triple quadrupole mass spectrometer with an electrospray ionization source (ESI). The samples were analyzed in positive-ion mode using multiple reaction monitoring (MRM). Transition of the precursor to the product ion was as follows: m/z 348.3→62.1 for AEA; m/z 379.3→287.2 for 2-AG. The LODs were as follows: 2 pg/mL for AEA and 40 pg/mL for 2-AG. Analyses were performed in three independent experiments. Obtained results were normalized for milligrams of protein. Endocannabinoids concentrations are expressed as a percentage of the concentrations found in control cells (15.9 ± 0.7 and 238 ± 16 fmol/mg protein for AEA and 2-AG, respectively).

Gęgotek A., Bielawska K., Biernacki M., Zaręba I., Surażyński A, & Skrzydlewska E. (2017). Comparison of protective effect of ascorbic acid on redox and endocannabinoid systems interactions in in vitro cultured human skin fibroblasts exposed to UV radiation and hydrogen peroxide. Archives of Dermatological Research, 309(4), 285-303.

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Protein Quantification by TMT Labeling

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Protein digestion and TMT labeling were carried out following the manufacturer's instruction.³⁰ The labeled peptides were desalinated and fractionated by high pH reversed‐phase liquid chromatography using the 1290 UPLC system (Agilent). The fractions were separated by nanoscale liquid chromatography and analyzed by on‐line electrospray MS/MS. Tandem mass spectra were extracted by Proteome Discoverer software version 2.4.1.15 (Thermo Fisher Scientific). All MS/MS samples were analyzed using SEQUEST, which was set up to search the Uniprot database. The percolator algorithm was used to control peptide level false discovery rates lower than 1%. The normalization of total peptide amount was used to correct experimental deviation.

Chen J., Jin J., Gu W., Zhao Z., Yuan H., Zhou Y., Nagle D.G., Xi Q., Zhang X., Sun Q., Wu Y., Zhang W, & Luan X. (2023). Crizotinib‐based proteolysis targeting chimera suppresses gastric cancer by promoting MET degradation. Cancer Science, 114(5), 1958-1971.

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UPLC-MS Analysis of JMT Powder

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50 mg of JMT powder was extracted in the mixture of 400 μL methanol and 100 μL water. Ultra-high performance liquid chromatography/mass spectrometry (UPLC/MS) analysis was performed on a 1290 UPLC system (Agilent Technologies, Santa Clara, CA, United States) with an ACQUITY UPLC C18 column (Waters Corp, Milford, MA). The flow rate was 0.4 ml/min and the sample injection volume was 5 μL. The mobile phase consisted of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The linear elution gradient program was as follows: 0–3.5 min, 95–85% A; 3.5–6 min, 85–70% A; 6–6.5, 70–70% A; 6.5–12 min, 70–30% A; 12–12.5 min, 30–30% A; 12.5–18 min, 30–0% A; 18–25 min, 0–0% A; 25–26 min, 0–95% A; 26–30 min, 95–95% A. The MS data was acquired by Q Exactive Focus mass spectrometer (Thermo Fisher Scientific, Waltham, MA, United States) with a mass range of 100–1500 m/z.

Sun Q., Zhang R., Xue X., Wu Q., Yang D., Wang C., Yan B, & Liang X. (2021). Jinmaitong Alleviates Diabetic Neuropathic Pain Through Modulation of NLRP3 Inflammasome and Gasdermin D in Dorsal Root Ganglia of Diabetic Rats. Frontiers in Pharmacology, 12, 679188.

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Quantitative Analysis of Fructoselysine

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The determination of fructoselysine was carried out on An Agilent 1290 UPLC system combined with an Agilent 6460 triple-quadrupole mass spectrometry equipped with a Jet Stream ESI source (Agilent Technologies Inc. Santa Clara, CA, USA). A Phenomenex Synergi Hydro-RP column (250 mm × 2 mm, 4 μm, 80 Å; Phenomenex, Torrance, CA, USA) was used for the chromatographic separation. The binary mobile phase compositions were methanol and 0.1% (v/v) formic acid in water (30 : 70, v/v). The isocratic condition was applied at a flow rate of 0.2 mL min⁻¹. The temperature of the column oven was maintained at 25 °C and the injection volume used was set at 5 μL.
The identification and quantitation of fructoselysine was achieved using the positive ESI and MRM mode. The optimized parameters used for this analysis are presented in the ESI (Table S1†). The quantitation and qualification ions of fructoselysine were m/z 309 → 84 and m/z 309 → 147, respectively. The optimal settings of MS for fructoselysine detection are reported in the ESI (Table S2†). The total and extracted ion chromatogram of each specific transition is also shown in the ESI (Fig. S1†). The fructoselysine was calibrated by a standard curve (r² = 0.9990) built with fructoselysine standard gradients.

Wang Y., Hu H., McClements D.J., Nie S., Shen M., Li C., Huang Y., Chen J., Zeng M, & Xie M. (2019). Effect of fatty acids and triglycerides on the formation of lysine-derived advanced glycation end-products in model systems exposed to frying temperature. RSC Advances, 9(27), 15162-15170.

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Quantitative Proteome Analysis by TMTpro

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For peptide extraction, 120 μg of TPP protein samples (37°C, 44°C, 47°C, 50°C, 53°C, 56°C, 59°C) were incubated with 100 mM DTT and 200μl of 8 M urea for 1 h at 56°C, followed by 50 mM IAA buffer and 100 mM TEAB buffer incubation for 30 min at room temperature. Furthermore, proteins were digested with 2.5 μg trypsin (Promega) in 45 μl 50 mM TEAB buffer for 18 h at 37°C. Peptides were labelled by TMTpro 16plex Lable (Thermo Fisher Scientific) following the manufacturer's instructions, desalted using Sep‐Pak C18 column (Waters) and dried in vacuo. Peptides were redissolved in 10 mM ammonium formate pH 10.0, then separated into fifth fractions by high pH gradient separation (from 0% to 90% of 10 mM ammonium formate in 90% ACN for 66 min and held for 5 min) using 1290UPLC system (Agilent Corp.) connected to a BEH C18 column (2.1 × 150 mm, 1.7 μm, 300 Å; Waters). The flow rate was 250 μl/min, and column temperature was kept at 50°C. Each fraction was collected and dried in vacuo.

Jiang Z., Zhang L., Yao Z., Cao W., Ma S., Chen Y., Guang L., Zheng Z., Li C., Yu K, & Shyh‐Chang N. (2022). Machine learning‐based phenotypic screening for postmitotic growth inducers uncover vitamin D3 metabolites as small molecule ribosome agonists. Cell Proliferation, 55(5), e13214.

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UPLC-MS/MS Quantitation of Analytes

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An Agilent 1290 UPLC system was combined with an Agilent 6460 series MS/MS system (Agilent Technologies, Santa Clara, CA, United States) to quantify the analytes in the ESI-positive ionization mode. The mass spectrum conditions were as follows: a capillary voltage of 4,000 V; a gas flow rate of 10 L/min; a nebulizer of 30 psi; a gas temperature of 350°C; and a Delta EMV(+) of 200 V. Five microliters of the samples was injected into an Agilent SB-C₁₈ column (2.1 mm × 50 mm, 1.8 μm particles). The analytes were separated using the gradient elution method, and the mobile phase was composed of water (0.05% V/V formic acid) (A) and acetonitrile (B) at a flow rate of 0.4 mL/min (0–1 min, 10–10% B; 1–5 min, 10–90% B; 5–7 min, 90–90% B; and 7–7.1 min, B 90–10%; rebalance for 2 min). The multiple reaction-monitoring parameters are shown in Table 1.

Nie W., Yang Y., Li L., Ding Y., Chen X., Li M., He N., Ji G., Zhang Y., Kang P, & Zhang T. (2023). Comparison of pharmacokinetic profiles of seven major bioactive components in normal and non-alcoholic fatty liver disease (NAFLD) rats after oral administration of Ling-Gui-Zhu-Gan decoction by UPLC-MS/MS. Frontiers in Pharmacology, 14, 1174742.

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Untargeted Metabolomic Profiling of Cellular Activity

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The changes in cellular activity were studied using untargeted metabolomic profiling by HPLC–MS (high-performance liquid chromatography coupled to mass spectrometry). Chromatography was performed in the normal phase with established protocols using a Cogent Diamond Hydride HILIC 150 × 2.1 mm column in an Agilent 1290 UPLC system [28 (link)]. Mass spectrometry was performed using an Agilent 6538 Q-TOF. Metabolites with a median intensity of zero across all experimental groups were excluded from the analysis. Undetected remaining metabolites (i.e., intensity of zero) were replaced with a value of one half of the minimum peak intensity for statistical analyses. Statistical analysis was performed in Metaboanalyst. The data were first log-transformed and standardized. The metabolomic profiles between the experimental groups were compared using principal components analysis (PCA), hierarchical clustering, volcano plot analysis, and pathway enrichment analysis. Significance was assessed with false discovery rate corrections using a significance level of 0.05 selected a priori.

Fredrikson J.P., Brahmachary P.P., Erdoğan A.E., Archambault Z.K., Wilking J.N., June R.K, & Chang C.B. (2022). Metabolomic Profiling and Mechanotransduction of Single Chondrocytes Encapsulated in Alginate Microgels. Cells, 11(5), 900.

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Metabolomics and Lipidomics Profiling

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Metabolites and lipids were extracted from the samples (See Supplementary Data 1 for further details of methodology). For metabolomics studies, an ultraperformance LC (UPLC) Vanquish system (Thermo Fisher Scientific, Waltham, MA, USA) with a UPLC BEH Amide column (2.1 mm × 100 mm, 1.7 μm) coupled to a Q Exactive HFX mass spectrometer (Orbitrap MS, Thermo Fisher) was used for separation and subsequent analysis (see Supplementary Data 1 for more details). For lipidomics, a 1290 UPLC system (Agilent Technologies, Santa Clara, CA, USA), equipped with a Kinetex C18 column (2.1 x 100 mm, 1.7 μm, Phenomen) coupled to a Q Exactive HFX mass spectrometer (Orbitrap MS, Thermo Fisher), was used for separation and later analysis (see Supplementary Data 1 for more details).

Zhang L., Wen X., Hou Y., Yang Y., Song W., Zeng Y, & Sun J. (2022). Integrated metabolomics and lipidomics study of patients with atopic dermatitis in response to dupilumab. Frontiers in Immunology, 13, 1002536.

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