1290 infinity 2 lc

Manufactured by Agilent Technologies

Sourced in United States

The 1290 Infinity II LC is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative separations. It features a modular design and advanced technology to provide reliable and efficient liquid chromatography analysis.

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1290 Infinity II Series LC system Agilent 1290 Infinity II LC – 6545 Q‐ToF Mass spectrometers 1290 Infinity II LC-6545 Quadrupole-TOF Agilent 1290 Infinity II LC 1290 Infinity II – 6550 iFunnel QTOF LC-MS system Agilent 1290 Infinity II LC system Agilent 1290 Infinity II Accurate Mass Q-ToF LC/MS spectrometer 1290 Infinity II LC System 1290 Infinity LC 1290 Infinity II

10 protocols using 1290 infinity 2 lc

Analyzing V272M DBD-PAT Interaction by MS

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Protein buffer of V272M DBD was replaced by 200 mM ammonium acetate buffer (pH 7.0) using dialysis device. V272M DBD was mixed with PAT at a molar ratio of 1 : 1 and the mixture was divided into two same samples. The mass of the two samples were immediately determined, without an incubation step, by native MS and denatured MS, respectively. The mass of V272M DBD without PAT treatment was also determined in the same way. Native MS was carried out on a 1290 Infinity II LC coupled with a 6230 LC/TOF system equipped with an Agilent Jet Stream source. LC separation was obtained with PolyHYDROXYETHYL ATM column (200 3 2.1 mm, 5 mm, 200 A ˚). In denatured MS, a 1290 Infinity II LC coupled with a 6530 LC/QTOF system and an Agilent 300SB-C8 column (50 3 2.1 mm, 3.5 mm) were used for denatured protein determination. All experimental MS data of the samples were processed using Agilent MassHunter Qualitative Analysis 7.0 software. Experimental molecular weights (MW) were determined from MS peak maxima with average MW and standard deviations calculated from identified charge states.

Tang Y., Song H., Wang Z., Xiao S., Xiang X., Zhan H., Wu L., Wu J., Xing Y., Tan Y., Liang Y., Yan N., Li Y., Li J., Wu J., Zheng D., Jia Y., Chen Z., Li Y., Zhang Q., Zhang J., Zeng H., Tao W., Liu F., Wu Y, & Lu M. (2022). Repurposing antiparasitic antimonials to noncovalently rescue temperature-sensitive p53 mutations. Cell reports, 39(2).

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PBAT Hydrolysis Products Quantification

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Following incubation with PBAT, the supernatants were diluted with ice-cold methanol (1:1 v/v) in order to precipitate the enzymes and acidified to pH 3.0 with 1 M HCl. The samples were centrifuged at 18.213 x g (5427 R, Eppendorf) for 15 min at 4 °C, then filtered through 0.45 μm nylon syringe filters and thereafter analysed by a HPLC-DAD system consisting of a 1290 Infinity II LC (Agilent Technologies), coupled with a reversed phase column C18 (Poroshell 120 EC-C18 2,7 µm 3.0 × 150 mm), at a flow rate of 0.4 mL/min^{15 (link),19}. The PBAT hydrolysis products were separated using a nonlinear gradient according to Quartinello et al.^{2 (link)} with some modifications (i.e. solvent A: H₂O, solvent B: methanol, solvent C: formic acid; solvent C was kept at 10% constantly; 0–13 min 15% B; 13–30 min 15–40% B; 30–35 min 40–90% B; 35–46 min 90–15% B; 46–60 min 15% B) and detected with a photodiode array detector (Agilent Technologies) at the wavelength of 245 nm. The expected released molecules (Fig. 1a) bis(4-hydroxybutyl) terephtalate (BTaB), mono(4- hydroxybutyl) terephthalate (BTa) and terephthalic acid (Ta) were quantified using external calibration curves in the range 0.001–0.5 mM. Values from controls without mycelium added were subtracted from the results. The calculated concentrations were plotted on graph by using Origin Pro software v.9.5 (https://www.originlab.com/origin).

Tesei D., Quartinello F., Guebitz G.M., Ribitsch D., Nöbauer K., Razzazi-Fazeli E, & Sterflinger K. (2020). Shotgun proteomics reveals putative polyesterases in the secretome of the rock-inhabiting fungus Knufia chersonesos. Scientific Reports, 10, 9770.

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Targeted LC-MS/MS Analysis of Drugs

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Samples (10 μL) were chromatographed on a ZORBAX SSHD Eclipse Plus C18 column (3×50 mm, 1.8 μm, 959757–302; Agilent, Santa Clara, CA) with a guard column (2.1×5 mm, 1.8 μm, 821725–901; Agilent) using 1290 Infinity II LC (Agilent). Column temperature and the LC flow rate were set at 40°C and 0.4 ml/min. Initial chromatographic condition was maintained at 90% mobile phase A (water with 0.1% formic acid, v/v) and 10% mobile phase B (acetonitrile with 0.1% formic acid, v/v) for one min, then increased to 80% B by 3 min, then to 95% B by 4 min, and then returned to initial condition at 5 min until 8 min for sufficient equilibrium. Unmodified mobile phases A (water) and B (acetonitrile) were used for the analysis of Troglitazone.
MS/MS analyses were performed in positive ion mode (all test drugs except Troglitazone) or negative ion mode (Troglitazone) with an electrospray ionization (ESI) source using 6470 triple-quadruple MS (Agilent). The capillary voltage was set at 3500 V. The nebulizer gas pressure and gas temperature were set at 35 psi and 350°C, respectively. The MS/MS parameters for each test compound are summarized in Supplemental Table 1.

Sakolish C., Reese C.E., Luo Y.S., Valdiviezo A., Schurdak M.E., Gough A., Taylor D.L., Chiu W.A., Vernetti L.A, & Rusyn I. (2020). Analysis of Reproducibility and Robustness of a Human Microfluidic Four-Cell Liver Acinus MicroPhysiology System (LAMPS). Toxicology, 448, 152651.

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Spectroscopic Characterization of Organic Compounds

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Chemicals were obtained from Sigma-Aldrich and Carbosynth and were used without further purification except when noted. Solvents were used as received or passed over a drying column [N,N′-dimethylformamide (DMF) and tetrahydrofuran]. Nuclear magnetic resonance (NMR) spectra were recorded on either a 400-MHz Varian Mercury-Oxford or a Bruker AVANCE (600 MHz) spectrometer. ¹H chemical shifts are reported in δ values relative to tetramethylsilane and referenced to the residual solvent peak [CDCl₃, δ_H = 7.26 parts per million (ppm) and δ_C = 77.16 ppm; D₂O, δ_H = 4.79 ppm]. Coupling constants are reported in hertz. HPLC measurements were performed using an Agilent 1100 Series instrument. High-resolution mass spectra were recorded on an Agilent 6230 Series time-of-flight mass spectrometer coupled to an Agilent 1290 Infinity II LC system [HPLC column: Agilent, EclipsePlus C18, 50 mm by 2.1 mm, particle size (1.8 μm); ionization method: electrospray ionization (ESI)]. HPLC purification was performed on an Agilent 1260 Infinity instrument (HPLC column: Luna 5u C18, 250 mm by 21.2 mm).

Ernst S., Ecker F., Kaspers M.S., Ochtrop P., Hedberg C., Groll M, & Itzen A. (2020). Legionella effector AnkX displaces the switch II region for Rab1b phosphocholination. Science Advances, 6(20), eaaz8041.

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Optimized AAV Characterization Protocol

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All protocols documented were optimized from testing different parameters such as AAV titers, sample incubation in different acidic conditions of formic acid (FA) and TFA, times of sample incubation, varying the LC gradients for optimal elution of the AAV VP proteins at the desired of ∼3 min, column temperature, sample injection volume, and different serotypes, with reproducibility and robustness.
Agilent 1290 Infinity II LC: auto-sampler with 20-μL loop for both intact and digested analyses.
Agilent 6545 LC/QTOF: Dual-jet stream source. See Table 3 for LC setting details, and Table 4 for MS setting details.

Lam A.K., Zhang J., Frabutt D., Mulcrone P.L., Li L., Zeng L., Herzog R.W, & Xiao W. (2022). Fast and high-throughput LC-MS characterization, and peptide mapping of engineered AAV capsids using LC-MS/MS. Molecular Therapy. Methods & Clinical Development, 27, 185-194.

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Characterization of IAA Metabolism

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We incubated sodium
salt of IAA (2.5 mM; Sigma) (the salt was dissolved in 1× PBS,
pH 7.0) with tES, free HRP, and tES-HRP (1 μM final concentration)
at 37 °C for 1 h. The products of reacted IAA samples were characterized
on UPLC-QTOF systems (1290 Infinity II LC + Agilent 6550B QTOF). Chromatographic
separation was achieved with a linear gradient using mobile phase
A (0.1% formic acid in water) and B (0.1% formic acid in acetonitrile)
at a flow rate of 0.40 mL·min^–1 on a Phenomenex
Kinetex 2.6 μm XB-C18 100 Å 100 × 2.1 mm column maintained
at 30 °C with a total run time of 20 min. The electrospray negative
ionization (ESI−) mode was selected, and the mass/charge ratio
was acquired in the 50–400 m/z range. Reference mass solution containing purine (119.04 m/z) and HP-0285 (302.00 m/z) dissolved in 95% acetonitrile/5% water/0.02%
acetic acid was used for real-time TOF mass correction.

Sadeghi S., Masurkar N.D., Vallerinteavide Mavelli G., Deshpande S., Kok Yong Tan W., Yee S., Kang S.A., Lim Y.P., Kai-Hua Chow E, & Drum C.L. (2022). Bioorthogonal Catalysis for Treatment of Solid Tumors Using Thermostable, Self-Assembling, Single Enzyme Nanoparticles and Natural Product Conversion with Indole-3-acetic Acid. ACS Nano, 16(7), 10292-10301.

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Conotoxin Identification and Quantification

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Concentrated and pooled HPLC fractions were run on UPLC–QTOF systems (1290 Infinity II LC + Agilent 6550B QTOF). Chromatographic separation was achieved with a linear gradient using mobile phase A (0.1% formic acid in water) and B (0.2% formic acid in acetonitrile) at a flow rate of 0.40 mL/min on a Phenomenex Kinetex 2.6-μm XB-C18 100 Å 100 × 2.1 mm column with a total run time of 10 min. Electrospray-positive ionization (ESI+) mode was selected, and the mass/charge was acquired in the 100–1700 m/z range.
The chromatogram was obtained by extracting the ion count of the dominant m/z species of conotoxin (tolerating an error of 50 ppm) against acquisition time. The retention time is then compared against synthetic peptide standards to determine conformation and relative quantity of the conotoxins.

Sadeghi S., Deshpande S., Vallerinteavide Mavelli G., Aksoyoglu A., Bafna J., Winterhalter M., Kini R.M., Lane D.P, & Drum C.L. (2021). A general approach to protein folding using thermostable exoshells. Nature Communications, 12, 5720.

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DLBS1442 Metabolite Identification using LC-MS/MS

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DLBS1442 was obtained from DLBS (Cikarang, West Java, Indonesia). Liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) analysis was performed using Agilent 1290 Infinity II LC with 6545 quadrupole-time of flight (QTOF) MS Detector. Separation was performed using XTerra MS C18 with 3.0 × 50 mm, 3.5 µm column. Acetonitrile (A) and ultrapure water (B) were used as the solvent. At a flow rate of 0.6 ml/minute, a gradient chromatographic system was performed using 2% solvent A into 100% solvent A for 11 minutes, followed by an equilibration of 2% solvent A over the next 4 minutes. MassHunter Workstation (B.06.01) was used as the processing software. The acquisition of MS/MS detector was performed at positive ion mode using Dual Agilent Jet Stream Electrospray ionization as an ion source.
LC-MS/MS result was processed using MZmine-2.32 (Pluskal et al., 2010) , and the m/z of parent ion and fragmented ion was then collected. For compound prediction, we used METLIN at www.metlin.scripps.edu (Guijas et al., 2018) . The simple search mode was used for early screening using the m/z value of the precursor ion. Predicted compounds from simple search were then continued with fragment similarity search using the m/z value of the precursor and fragment ions. Other references such as journals related to the metabolite compounds of DLBS1442 were also used in this metabolomics study.

Rahardjo N., Ramdani E.D., Tjandrawinata R.R., & Tjandrawinata R.R. (2020). Metabolomic study and in silico approach of DLBS1442 as progesterone receptor agonist. Journal of Applied Pharmaceutical Science, 10(5), 63-69.

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Peptide Mapping Protocol for LC-MS

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The same samples were also analyzed on a standard-flow platform (Agilent 1290 Infinity II LC). Ten microliters of digest was directly injected onto a 25-cm column (AdvanceBio Peptide Mapping, C18, 2.1 mm×250 mm, 2.7um, 120 Å) and separated over a 23-minute gradient at 350 μL/min (0-0.5 min, 5%-7.5% B; 0.

Yin X., Baig F., Haudebourg E., Blankley R.T., Gandhi T., Müller S., Reiter L., Hinterwirth H., Pechlaner R., Tsimikas S., Santer P., Willeit J., Kiechl S., Witztum J.L., Sullivan A, & Mayr M. (2017). Plasma Proteomics for Epidemiology: Increasing Throughput With Standard-Flow Rates. Circulation. Cardiovascular genetics, 10(6).

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Quantifying Phenylpropanoid Metabolites by UHPLC-MS/MS

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Contents of phenylpropanoids were assessed according to Cotrozzi et al. (2018b) , with some modifications. Samples (30 mg FW) were homogenized in 500 µL of 80% HPLC-grade methanol [in water (v/v) ]. The supernatant was diluted five-fold with an aqueous solution of 0.2% formic acid. UHPLC-ESI-MS/MS analyses were performed on an Agilent 1290 Infinity II LC system coupled to a 6495 Triple Quadrupole mass spectrometer equipped with a Jet Stream electrospray (ESI) ionization source (Agilent Technologies, Santa Clara, CA, USA). The separation was achieved at 35 °C using a reverse-phase Agilent column (Zorbax Eclipse Plus, C18, 1.8 µm particle size, 2.1 mm i.d., 50 mm length). A detailed description of analytical conditions is available in Assumpção et al. (2018) . In the present study, phenylpropanoid metabolites are grouped and presented as total free phenolic acids (Tot Phen) and flavonoids (Tot flav) on the basis of their chemical structure.

Pellegrini E., Hoshika Y., Dusart N., Cotrozzi L., Gérard J., Nali C., Vaultier M.N., Jolivet Y., Lorenzini G, & Paoletti E. (2019). Antioxidative responses of three oak species under ozone and water stress conditions. The Science of the total environment, 647.

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