Waters 996 pda

Manufactured by Waters Corporation

Sourced in Ireland, United States

The Waters 996 PDA is a photodiode array (PDA) detector designed for high-performance liquid chromatography (HPLC) applications. It provides rapid and sensitive detection of a wide range of compounds across the UV-Vis spectrum. The 996 PDA offers flexible wavelength selection and high-speed data acquisition to support a variety of analytical needs.

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Lab products found in correlation

Masslynx 4, by Waters Corporation (2 mentions) Micromass quattro micro triple quadrupole, by Waters Corporation (2 mentions) Waters alliance 2695, by Waters Corporation (2 mentions) Waters 600, by Waters Corporation (1 mentions) Dmso d6, by Cambridge Isotopes (1 mentions) Luna c18 2, by Phenomenex (1 mentions) Waters 1525 pump, by Waters Corporation (1 mentions) Vns600, by Agilent Technologies (1 mentions)

5 protocols using waters 996 pda

HPLC-DAD-MS/MS Analysis of Compound Separation

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Samples were also analysed by an HPLC-DAD-MS/MS system: a Waters Alliance 2695 (Waters^®, Dublin, Ireland) separation module with an autosampler (20 μL injection volume), a quaternary pump and a solvent degasser, coupled to a Photodiode Array Detector Waters 996 PDA (Waters, Dublin, Ireland) scanning wavelength absorption between 210 and 600 nm. A LiChrospher^® 100 RP-18 5 μm column at 35 °C (stabilized by a column oven) was used. Tandem mass spectrometry (MS/MS) detection was carried out with a Micromass^® Quattro Micro triple quadrupole (Waters, Dublin, Ireland), using an electrospray ionization source in both positive (ESI+) and negative (ESI-) modes. A full scan mode (m/z: 60–1100) record was applied for the mass spectra of the compounds separated by HPLC, using a collision energy of 20 eV. The HPLC gradient method and eluents are described in ‘Supplementary Materials Table S2’ section. The MS/MS conditions, as source temperature, capillary and source voltages have been previously described by Katsinas et al. [16 (link)]. For data acquisition and processing, MassLynx^® 4.1 software (Waters, Dublin, Ireland) was used.

Hernández F., Andreu-Coll L., Bento-Silva A., Serra A.T., Mena P., Legua P, & Bronze M.R. (2022). Phytochemical Profile of Opuntia ficus-indica (L.) Mill Fruits (cv. ‘Orito’) Stored at Different Conditions. Foods, 11(2), 160.

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HPLC Analysis of Urinary Hypoxanthine and Creatinine

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Urinary hypoxanthine and creatinine concentrations were determined using an adaptation of the high-performance liquid chromatography (HPLC) method described by George et al.³¹ Briefly, urine samples were thawed and sonicated before 200 μL was transferred to an Eppendorf tube containing 1 × 10⁻⁷ mol of 2-aminopurine (internal standard). The samples were then analyzed on an HPLC (Waters 996 PDA, Waters 600 controller, and 717plus autosampler; Millipore Corp) by injecting 35 μL onto a Supelcosil LC-18-S 15 cm × 4.6 mm, 5 μm column (SGE; Austin, TX), with the following isocratic conditions: 10 mM potassium dihydrogen phosphate buffer, pH 4.7, flow rate 1.0 mL/min. Creatinine, hypoxanthine, and 2-aminopurine were quantitated by obtaining peak areas at the appropriate retention times (~3.5, 8, and 13.5 minutes, respectively) and wavelengths (230, 248, and 305 nm, respectively). The area ratios of each compound to 2-aminopurine were determined and converted into concentration using standard curves. Samples were analyzed in triplicate and values with a coefficient of variation less than 10% were included in the final analysis. The limits of detection were 1.58 μM for hypoxanthine and 3.2 μM for creatinine.

Holden M.S., Hopper A., Slater L., Asmerom Y., Esiaba I., Boskovic D.S, & Angeles D.M. (2014). Urinary Hypoxanthine as a Measure of Increased ATP Utilization in Late Preterm Infants. Infant, child & adolescent nutrition, 6(4), 240-249.

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HPLC-DAD-MS/MS Identification of Phenolic Compounds

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The phenolic compounds present in the freshly prepared aqueous solutions of CONV and OPT3, and the degradation phenolic products present in the 40 °C exposed aqueous solutions of OL, HT, CONV, and OPT3, were putatively identified by an HPLC-DAD-MS/MS system: Waters Alliance 2695 (Waters^®, Dublin, Dublin, Ireland) separation module with an autosampler (10 μL injection volume), a quaternary pump and a solvent degasser, coupled to a Photodiode Array Detector Waters 996 PDA (Waters^®, Dublin, Ireland) scanning wavelength absorption between 210 and 600 nm. A LiChrospher^® 100 RP-18 5 μm (250 × 4.0 mm) (Sigma-Aldrich, St. Louis, MO, USA) column at 35 °C (stabilized by a column oven) was used. MS/MS detection was carried out with a Micromass^® Quattro Micro triple quadrupole (Waters^®, Dublin, Ireland), using an ESI. A full scan mode (m/z: 60–1100) record was applied for the mass spectra of the compounds separated by HPLC, using a collision energy of 20 eV. The HPLC gradient method, eluents, and elution program used, together with the source temperature, capillary, and source voltages are described by Katsinas et al. [16 (link)] For data acquisition and processing, MassLynx^® 4.1 software (Waters^®, Dublin, Ireland) was used.

Katsinas N., Enríquez-de-Salamanca A., Bento da Silva A., Bronze M.R, & Rodríguez-Rojo S. (2021). Olive Pomace Phenolic Compounds Stability and Safety Evaluation: From Raw Material to Future Ophthalmic Applications. Molecules, 26(19), 6002.

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Isolation and Characterization of Bioactive Compounds

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Isolation of extracts was conducted by binary HPLC pump (WATERS 600, Waters Co., Milford, MA, USA) and UV-vis detector (WATERS 996 PDA, Waters Co.) was conducted using reversed-phase Phenomenex Luna 5 μ C₁₈ column (250 × 10.00 mm, 5 μm, Waters Co.) at a flow rate of 2.0 mL/min. Low-resolution LC-MS data were measured using an HPLC (Agilent Technologies, Santa Clara, CA, USA) with a reversed-phase Phenomenex Luna 5μm C₁₈ column (4.6 × 100 mm, 5 μm) at a low flow rate of 1.0 mL/min. NMR spectra were recorded on 300 and 125 MHz for ¹H and ¹³C NMR (Bruker Advance, Billerica, MA, USA), respectively, using the solvent DMSO-d₆ (Cambridge Isotope Laboratories, Inc. Tewksbury, MA, USA). EP grade solvents (Dae-Jung Chemicals & Metals Co. Ltd, Seoul, Korea) were used for fractionation of extracts. For HPLC analyses and LC-MS, HPLC grade solvents were used (J.T.Baker and Dae-Jung Chemicals & Metals Co. Ltd.).

Kim K., Jeong H.I., Yang I., Nam S.J, & Lim K.M. (2019). Acremonidin E produced by Penicillium sp. SNF123, a fungal endophyte of Panax ginseng, has antimelanogenic activities. Journal of Ginseng Research, 45(1), 98-107.

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NMR Spectroscopy and HPLC Analysis Protocol

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The ¹H and ¹³C NMR spectra were recorded at 25 °C using a 600 MHz Fourier transform nuclear magnetic resonance device (VNS600; Agilent Technologies, Santa Clara, CA, USA) at the Core Research Support Center for Natural Products and Medical Materials (CRCNM) in the Yeungnam University. Medium pressure liquid chromatography (MPLC) was also conducted using Biotage Isolera (Biotage, Uppsala, Sweden). In addition, high-performance liquid chromatography (HPLC) was conducted on a Waters system (Waters 1525 pump and Waters 996 PDA; Waters, Milford, MA, USA) with a semi-preparative HPLC column (Phenomenex Luna C18(2), 10 mm × 250 mm, 5 μ (Phenomenex, Torrance, CA, USA) and an analytical HPLC column (Phenomenex Luna C18(2), 4.6 mm × 250 mm, 5 μ (Phenomenex, Torrance, CA, USA).

Chae H.J., Kim G.J., Deshar B., Kim H.J., Shin M.J., Kwon H., Youn U.J., Nam J.W., Kim S.H., Choi H, & Suh S.S. (2021). Anticancer Activity of 2-O-caffeoyl Alphitolic Acid Extracted from the Lichen, Usnea barbata 2017-KL-10. Molecules, 26(13), 3937.

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