Agilent 1260 infinity hplc

Manufactured by Agilent Technologies

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The Agilent 1260 Infinity HPLC is a high-performance liquid chromatography system designed for analytical applications. It provides precise and reliable separation and quantification of a wide range of chemical compounds.

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69 protocols using agilent 1260 infinity hplc

Lauric Acid Quantification in CKE

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The sample preparation procedure was simple [49 ,50 ]. A 1.0 g of CKE was combined with 1 ml of HPLC-grade methanol that was purchased from the Merck company in Mumbai, India. Next, a 0.45 μm filter was used to filter the mixture. The standard graph of LA was obtained using pure LA (Merck, Mumbai, India).
The Agilent 1260 Infinity HPLC (analytical) system was utilized to ascertain the Lauric acid content in CKE. The apparatus consisted of an SPD-10Avp ultraviolet detector, an LC-10ADvp pump, a SIL-10ADvp auto sampler, a DGU-14A degasser, and a SCL-10Avp system controller. The stationary C18 column was operated at room temperature. The mobile phase was HPLC-grade methanol, flowing at a rate of 0.5 ml/min. The injection volume for every chromatographic run was 50 μl. A wavelength of 203 nm was used for the detection process. Next, we used the Agilent 1260 Infinity HPLC (preparative) system under the same conditions to isolate the targeted peak that we had obtained from CKE. The HPLC analysis was performed under the supervision of Guwahati Biotech Park, Guwahati, Assam.

Sandhya S., Talukdar J., Gogoi G., Dey K.S., Das B, & Baishya D. (2024). Impact of coconut kernel extract on carcinogen-induced skin cancer model: Oxidative stress, C-MYC proto-oncogene and tumor formation. Heliyon, 10(8), e29385.

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HPLC Analysis of Volatile Fatty Acids

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The levels of volatile fatty acids present in the supernatant of both co-cultures and monocultures were measured using an Agilent1260 Infinity HPLC (Agilent) system. The samples were prepared by acidifying to 5 mM, using sulfuric acid, and then incubating at room temperature for 5 min. Samples were then centrifuged for 5 min at 21,000 g. The supernatant was syringe filtered into an HPLC vial (Eppendorf FA-45–24–11) using a 0.22-µm polyvinylidene difluoride (PVDF) filter. The samples were analyzed using an Agilent 1260 Infinity HPLC (Agilent) system equipped with an autosampler unit (1260 ALS). The separation of formate, acetate, glucose, and lactate was carried out using a Bio-Rad Aminex^® 87H Ion Exclusion Column for organic acids (Part No. 1250140; Bio-Rad Laboratories, Inc., Hercules, CA, USA), with a mobile phase of 5 mM sulfuric acid. In-house standards were prepared with MC-blank culture medium as a base and sodium formate (ACS Grade, Fisher Chemical S648500), sodium acetate (ACS Grade, Fisher Chemical S210500), and L-lactic acid sodium (99%, extra pure; Acros Organics, 439220100), and D-(+)-glucose (Sigma-Aldrich Cat. No. G8270) at concentrations of 0.1 g/L and 1 g/L.

Brown J.L., Gierke T., Butkovich L.V., Swift C.L., Singan V., Daum C., Barry K., Grigoriev I.V, & O’Malley M.A. (2023). High-quality RNA extraction and the regulation of genes encoding cellulosomes are correlated with growth stage in anaerobic fungi. Frontiers in Fungal Biology, 4, 1171100.

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Extraction and Purification of XAN from X. sorbifolia

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The husks of X. sorbifolia were collected from Chifeng, the Nei Mongol Autonomous Region, China (lat. 41°17′10″ N, long. 116°21′07″E; altitude 300 m a.s.l.), in a dry season Oct 2011. A voucher specimen (NO.WGG-1110) was deposited in the Department of Natural Products Chemistry, Shenyang Pharmaceutical University, China. XAN was separated from 10.0 kg of husks according to our previous study (Meng et al., 2016 (link)), which was dissolved in distilled water (1% DMSO) for oral administration by gavage. The HPLC system (Agilent HPLC-1260 Infinity, Santa Clara, California, USA; Diamonsil C18 column, 250 mm × 4.6 mm, 5 µm, Beijing DIMAGIT Science & Technology Co., Ltd., Beijing, China; UV detector 210 nm) revealed that the purity of XAN was higher than 99.0% as shown in Figure S2 (Supplementary Files).
Aβ protein fragment 1–42 (Aβ_1–42) was purchased from Sigma–Aldrich (St. Louis, MO, USA). Donepezil hydrochloride tablets was purchased from Eisai China Inc. (Tokyo, Japanese). Acetonitrile and formic acid of HPLC grade were purchased from Tedia (Fairfield, OH, USA) and Dikma Corp. (Richmond Hill, NY, USA).

Zhou H., Tai J., Xu H., Lu X, & Meng D. (2019). Xanthoceraside Could Ameliorate Alzheimer’s Disease Symptoms of Rats by Affecting the Gut Microbiota Composition and Modulating the Endogenous Metabolite Levels. Frontiers in Pharmacology, 10, 1035.

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Quantification of Macrolide Antibiotics in Biomass

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Biomass was assayed by measuring the optical density at 600 nm (OD₆₀₀). Concentration of AVM and its derivatives (IVM, doramectin and emamectin) and other macrolide antibiotics was determined by Agilent HPLC-1260 Infinity (Agilent, USA) using a Unitary C18 column (250 mm × 4.6 mm × 5 µm) (Acchrom, China) as previous research (Wang et al. 2015 (link)). Briefly, AVM, IVM, doramectin and emamectin were monitored at 245 nm with acetonitrile-water (90:10, v/v) at 2.0 mL/min. Other macrolides were monitored at 215 nm with K₂HPO₄ buffer (0.05 M, pH 8.2)-acetonitrile (50:50, V/V) at 1.0 mL/min. The data was analyzed with analysis of variance (ANOVA) statistics with SPSS (IBM, USA).

Wang Y., Gong M., Wang X., Peng X., Wang Y., Guan J., Cheng D., Weng C, & Zheng Y. (2020). Efficient degradation of ivermectin by newly isolated Aeromonas taiwanensis ZJB-18,044. Biodegradation, 31(4-6), 275-288.

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HPLC Analysis of Compound LE

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Agilent 1260 Infinity HPLC (Agilent Technologies, Stockport, UK) was used for analysis. LE was separated on a C-18 column (250 mm × 5 mm × 4.6 mm). The mobile phase consisted of 0.04% formic acid in water (A) and acetonitrile (ACN) in 0.04% formic acid (B). An injection volume of 50 µL was used with the gradient conditions given in Table 1. The column temperature was set at 25 °C and the sample absorbance at 278 nm was monitored throughout each run.

Tejpal S., Wemyss A.M., Bastie C.C, & Klein-Seetharaman J. (2020). Lemon Extract Reduces Angiotensin Converting Enzyme (ACE) Expression and Activity and Increases Insulin Sensitivity and Lipolysis in Mouse Adipocytes. Nutrients, 12(8), 2348.

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Enzymatic Activity Monitoring of Recombinant UGMs

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The enzymatic activity of each recombinant UGM was monitored using an HPLC-based assay.^{10 (link),52 (link)} Small molecules were added from DMSO stocks to a final concentration of 1% DMSO. Reactions were analyzed using a CarboPac PA-100 column (Thermo Scientific) on an Agilent 1260 Infinity HPLC using an isocratic elution.

Kincaid V.A., London N., Wangkanont K., Wesener D.A., Marcus S.A., Héroux A., Nedyalkova L., Talaat A.M., Forest K.T., Shoichet B.K, & Kiessling L.L. (2015). Virtual Screening for UDP-Galactopyranose Mutase Ligands Identifies a New Class of Antimycobacterial Agents. ACS chemical biology, 10(10), 2209-2218.

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Quantifying Urinary Quinolinic Acid via LC-MS/MS

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All sample aliquots (approximately 4ml of urine in a 5ml cryovial) were shipped on dry ice in a single batch for analysis to the laboratories at Ethos Research & Development, Newport KY for quantifying QA in urine. As described previously (Nassan, Gunn, et al., 2019 (link)), Calibrators and internal standards for QA were created from solid standards. Bio rad lyphocheck quantitative urine levels 1 and 2 were used as quality control (QC) samples. Calibrators, QC samples, and patient samples were all prepared in the same manner. QA was analyzed using LC-MS/MS (Agilent 1260 Infinity HPLC and Agilent 6410B triple quadrupole mass spectrometer). QA samples were resolved on Agilent Poroshell 120 EC-C18, 2.7 μm, 3.0 × 100 mm column. QA was quantified using Mass Hunter Quantitative Analysis Version B.08.00 Software.

Nassan F.L., Gunn J.A., Hill M.M., Williams P.L, & Hauser R. (2020). Association of urinary concentrations of phthalate metabolites with quinolinic acid among women: a potential link to neurological disorders. Environment international, 138, 105643.

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Yeast Strains Cider Fermentation Protocol

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Yeast starters were incubated in liquid YPD overnight at 30 °C with a rotation speed of 220 rpm. Yeast cells were collected by centrifugation and suspended in sterile water. For each yeast, cider fermentations were conducted in triplicate with 500 mL-flasks containing 400 mL apple juice supplemented with 50 mg/L potassium metabisulfite. The fermentations were initiated by adding yeast starters at a final optical density (OD_600nm) of 0.2 and maintained at 30 °C under static conditions until the total soluble solids decreased to approximately 8.0 °Brix. The fermentation periods were 66, 72 and 216 h for the AQ, SC and KM strains, respectively. The fermented samples were centrifuged at 10,000 rpm for 10 min to separate the supernatants and yeast cells for further biochemical, transcriptional and metabolomic analyses. Total titratable acids in the ciders were determined by titration with NaOH and represented as g/L malic acid. The colour index was analysed by a UV–Vis spectrophotometer at a wavelength of 420 nm. The contents of ethanol, glucose, sucrose and fructose were quantified on an Agilent 1260 Infinity HPLC (Agilent Technologies, Santa Clara, CA, USA) fitted with a refractive index detector using a MetaCarb 87H column (300 mm × 4.6 mm). The column temperature was set at 35 °C, and 5 mM H₂SO₄ (pH 2.0) was used as the mobile phase at a flow rate of 0.6 mL/min.

Zhang Z., Lan Q., Yu Y., Zhou J, & Lu H. (2022). Comparative metabolome and transcriptome analyses of the properties of Kluyveromyces marxianus and Saccharomyces yeasts in apple cider fermentation. Food Chemistry: Molecular Sciences, 4, 100095.

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Benzene Oxidation Reaction Protocol

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Benzene oxidation reaction was carried out in a 50-ml Teflon-lined stainless steel reactor with 0.4 ml of benzene, 6 ml of H₂O₂ (30%), and 3 ml of CH₃CN at 25° or 0°C. After the reaction, an additional 20 ml of CH₃CN was added to transfer the products and 0.2 ml of toluene was also added as an internal standard. The products were analyzed with Agilent 1260 Infinity HPLC using a Unitary C-18 column. Before the analysis, the products were filtered by a syringe with a filter head.

Deng D., Chen X., Yu L., Wu X., Liu Q., Liu Y., Yang H., Tian H., Hu Y., Du P., Si R., Wang J., Cui X., Li H., Xiao J., Xu T., Deng J., Yang F., Duchesne P.N., Zhang P., Zhou J., Sun L., Li J., Pan X, & Bao X. (2015). A single iron site confined in a graphene matrix for the catalytic oxidation of benzene at room temperature. Science Advances, 1(11), e1500462.

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HPLC Analysis of PTS and PTSO in Agar

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For the analysis of PTS and PTSO in agar samples, an Agilent 1260 Infinity HPLC (Agilent Technologies Inc., Waldbron, Germany) system was used. The system is equipped with an online degasser, an autosampler, a column thermostat, a diode array detector, and a quaternary pump. The technology used to determine PTS and PTSO was previously described by our group [50 (link),51 (link)]. The analysis was carried out in a C18 column (Zorbax Eclipse Plus 50 mm × 4.6 mm, 1.8 μm). Solvents used were 30 mM perchloric acid and MeCN (solvent A and B, respectively) dissolved in water at a flow rate of 0.85 mL min⁻¹. The injection volume was 10 μL and the gradient elution program was: 0 min, 50% B; 2.2 min, 50% B; 4.5 min, 100% B; 6.8 min, 100% B; 8 min, 50% B; 10.5 min, 50% B. The wavelength of detection was set at 200 nm. Agar samples were individually weighed and extracted in 500 μL of methanol through 5 min in vortex. The extract was filtered and directly injected into the HPLC-UV.

Sorlozano-Puerto A., Albertuz-Crespo M., Lopez-Machado I., Gil-Martinez L., Ariza-Romero J.J., Maroto-Tello A., Baños-Arjona A, & Gutierrez-Fernandez J. (2020). Antibacterial and Antifungal Activity of Propyl-Propane-Thiosulfinate and Propyl-Propane-Thiosulfonate, Two Organosulfur Compounds from Allium cepa: In Vitro Antimicrobial Effect via the Gas Phase. Pharmaceuticals, 14(1), 21.

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