Model 515

Manufactured by Waters Corporation

Sourced in United States

The Waters Corporation Model 515 is a high-performance liquid chromatography (HPLC) pump. It is designed to provide precise and reliable solvent delivery for a wide range of analytical applications.

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13 protocols using model 515

HPLC Assay for Drug Quantification

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The apparatus used was a Waters HPLC system model 746 (Milford, US), consisting of a model 515 intelligent solvent delivery pump, a 100 μl injection loop, a computerized system controller, and a Waters 2487 UV detector. HPLC assay was carried out on a μ-Bondapack C₁₈ column (250 mm × 4.6 mm, 10 μm, Waters, Ireland). The mobile phase consisted of potassium dihydrogen phosphate (0.1 M)/acetonitrile at 80/20 (pH, 3 ± 0.1) eluted at flow rate 1.5 ml/min. Column effluent was detected at 276 nm with a UV detector. Column temperature was set at 40°C, and 50 µl of samples was injected to the HPLC system.
Quantitation was achieved by measurement of the peak area ratios of the drug to the IS.

Emami J, & Rezazadeh M. (2016). A simple and sensitive high-performance liquid chromatography method for determination of ciprofloxacin in bioavailability studies of conventional and gastroretentive prolonged-release formulations. Advanced Biomedical Research, 5, 163.

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Pressurized Liquid Extraction of Samples

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The PLE assays were carried out in an extraction unit developed at LATESC and minutely described by Gonçalves Rodrigues et al (2019) (link).
First, 5 g of sample was inserted into the 90 mL extraction vessel and its void fraction was filled with cotton layers and glass beads. Following this, the solvent was delivered to the system by an HPLC pump (Waters, Model 515, Milford, MA, USA), passed through a pre-heating vessel to reach the target temperature, and then flowed downstream through the extraction vessel. Once the extraction pressure of 100 bar was reached, the needle valve was opened and the extraction began in a continuous mode. During the extraction process, 100 mL of solvent were used, maintaining the same 1:20 w/v solute to solvent ratio used for MAE. The specific PLE variables can be seen in section 2.7.
Finally, the extracts were submitted to rotary evaporation and stored in amber flasks at −18 °C until further analysis.

Chada P.S., Santos P.H., Rodrigues L.G., Goulart G.A., Azevedo dos Santos J.D., Maraschin M, & Lanza M. (2022). Non-conventional techniques for the extraction of antioxidant compounds and lycopene from industrial tomato pomace (Solanum lycopersicum L.) using spouted bed drying as a pre-treatment. Food Chemistry: X, 13, 100237.

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HPLC Analysis of Compound Mixtures

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Waters HPLC (Milford, MA, USA) system used was equipped with a binary gradient pump (Model 515, Waters), an autosampler injector (Model 2707, Waters) and a diode array detector (Model 2998, Waters). Waters HPLC interface and Empower software by Waters corporation, USA was utilized for data acquisition. The analytical column RP-18e B Lichrosphere^® (250 mm × 4 mm, 5 μm, Merck, Germany) at 30°C ± 3°C was used for the study. The mobile phase included ACN water (70:30, v/v) and was degassed before analysis using a Millipore vacuum pump. Column effluent was monitored at 220 nm and 254 nm (photodiode array detection) with a total runtime of 30 min. The flow rate was maintained 1.0 ml/min. and injection volume remained 20 μL.

Naqvi A., Malasoni R., Gupta S., Srivastava A., Pandey R.R, & Dwivedi A.K. (2017). In Silico and In Vitro Anticancer Activity of Isolated Novel Marker Compound from Chemically Modified Bioactive Fraction from Curcuma longa (NCCL). Pharmacognosy Magazine, 13(Suppl 3), S640-S644.

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Characterization of High-G/M and Mid-G/M Alginates

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Sodium alginate with molecular weight (M_w) of 520 kDa and G/M ratio of 64/36 was designated High-G/M alginate, and sodium alginate with M_w of 532 kDa and G/M ratio of 34/66 was designated Mid-G/M alginate. Both were purchased from Qingdao Mingyue Seaweed Group Co., Ltd. (Qingdao, China). The detailed information of both purchased alginates was characterized by GPC and ¹HNMR spectroscopy (Table 1).
The G/M ratio of the respective alginates was determined at 80°C by ¹HNMR spectroscopy using an AVANCE NB-360 spectrometer (Bruker, Germany). The M_w and molecular weight distribution (M_w/M_n) of alginate samples were analyzed via gel permeation chromatography (GPC) using a system equipped with two TSK-gel size-exclusion columns (G4000PWXL and G3000PWXL), a refractive index detector (Waters, model 2414, Milford, MA), and an HPLC pump (Waters, model 515).
Collagen, acquired from rat tails, and was obtained by our laboratory. Anhydrous calcium chloride (CaCl₂) was purchased from Sigma-Aldrich (Merck Life Science (Shanghai) Co., Ltd.). All other chemical reagents were analytical grade and used as received.

Zheng G., Xue C., Cao F., Hu M., Li M., Xie H., Yu W, & Zhao D. (2023). Effect of the uronic acid composition of alginate in alginate/collagen hybrid hydrogel on chondrocyte behavior. Frontiers in Bioengineering and Biotechnology, 11, 1118975.

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HPLC Analysis of Withanolide A

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chromatographic conditions: The Waters HPLC (Milford, MA, USA) system used was equipped with a binary gradient pump (model 515, Waters), an autosampler injector (Model 2707, Waters), and a diode array detector (Model 2998, Waters). The Waters HPLC interface and Empower 2 software were utilized for data acquisition. The HPLC resolution of Withanolide A was achieved on an RP-18e Lichrosphere^® column (250 × 4 mm, 5 μm, Merck, Germany) at 30 ± 3°C utilizing the mobile phase consisting of acetonitrile: Water (1:1); flow rate of 1.0 mL/minute, and injection volume that remained at 20 μL. The column effluent was monitored at 240 nm (PDA detection).

Ahmad H., Khandelwal K., Pachauri S.D., Sanghwan R.S, & Dwivedi A.K. (2014). Stability indicating studies on NMITLI 118RT+ (standardized extract of withania somnifera dunal). Pharmacognosy Magazine, 10(39), 227-233.

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HPLC Analysis with Multidetector Setup

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A Waters HPLC system consisted of an isocratic pump (Model 515), an autosampler (Model 710 plus) and a variable UV-Vis detector (Model 480) was used. A multi-channel Chrom & Spec software for chromatography, version 1.5 x was used for data processing. A dry air oven (Melag, Germany) was used for heating the samples. The light sources were a l00 W Tungsten lamp (visible light) and a low-pressure Mercury lamp 200 W (UV light) with λ_max around 254 nm.

Souri E., Aghdami A.N, & Adib N. (2014). A stability indicating HPLC method for determination of mebeverine in the presence of its degradation products and kinetic study of its degradation in oxidative condition. Research in Pharmaceutical Sciences, 9(3), 199-206.

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Fluorescent Particle Microscopy Analysis

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Fluorescent polystyrene particles (0.79, 0.92 and 1.0 µm, Thermo Scientific Fluoro-Max) were suspended in deionized water (with 0.1% of Triton X to reduce agglomeration) in a concentration of ∼ 0.001 vol%.
An HPLC pump (Waters, model 515) was used to pump the samples through the devices at a controlled flow rate with a read out of the pressure.
During the operation, the devices were observed with an inverted fluorescence microscope (Olympus IX73 with an Orca-Flash 4.0 LT digital CMOS camera). Images were taken with a magnification of 20X and a 2 s exposure time.

Cruz J, & Hjort K. (2021). High-resolution particle separation by inertial focusing in high aspect ratio curved microfluidics. Scientific Reports, 11, 13959.

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HPLC Analysis of Organic Compounds

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The apparatus used was a Waters HPLC system model 746 (Milford, US) consisting of a model 515 intelligent solvent delivery pump, a 100-μL injection loop, a computerized system controller, and a Waters 2487 UV detector. Chromatographic separation was achieved using a μ-Bondapak C18 column (3.9 mm × 250 mm Waters, Ireland). The mobile phase consisted of water/acetonitrile at 60/40 eluted at a flow rate of 1.2 mL/min. Column effluent was detected at 230 nm with a UV detector.

Kazemi M., Emami J., Hasanzadeh F., Minaiyan M., Mirian M, & Lavasanifar A. (2020). Development of a RP-HPLC method for analysis of docetaxel in tumor-bearing mice plasma and tissues following injection of docetaxel-loaded pH responsive targeting polymeric micelles. Research in Pharmaceutical Sciences, 15(1), 1-13.

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HPLC Analysis of Thermal and Photolytic Degradation

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The HPLC system consisted of a pump (Model 515), a UV-Vis detector (Model 486), and an autosampler (Model 717) from Waters (Milford, USA). The HPLC data were processed using multi-channel Chrom&Spec software, version 1.5 x. Thermal degradation was performed in a Melag dry air oven (Germany). A 100 W tungsten (visible light) and a low pressure mercury lamp (UV light) were used for the photolytic degradation studies.

Souri E., Zargarpoor M., Mottaghi S., Ahmadkhaniha R, & Kebriaeezadeh A. (2014). A Stability-Indicating HPLC Method for the Determination of Fingolimod in Pharmaceutical Dosage Forms. Scientia Pharmaceutica, 83(1), 85-93.

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HPLC Analysis of Acetate Buffer Compounds

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The apparatus used was a Waters HPLC system model 746 (Milford, US) consisting of a model 515 intelligent solvent delivery pump, a 100 μl injection loop, a computerized system controller, and a Waters 2487 UV detector. Chromatographic separation was achieved using a µ-Bondapak C 18 column (3.9 mm × 250 mm Waters, Ireland) heated at 58 • C. The mobile phase consisted of acetate buffer (0.01 M)/ acetonitrile at 58/42 (pH, 5 ± 0.1) eluted at flow rate 1.9 ml/min. Column effluent was detected at 227 nm with a UV detector.

Rezazadeh M., Emami J., Mostafavi A., Rostami M., Hassanzadeh F., Sadeghi H., Minaiyan M, & Lavasanifar A. (2015). A Rapid and Sensitive HPLC Method for Quantitation of Paclitaxel in Biological Samples using Liquid-Liquid Extraction and UV Detection: Application to Pharmacokinetics and Tissues Distribution Study of Paclitaxel Loaded Targeted Polymeric Micelles in Tumor Bearing Mice. Journal of pharmacy & pharmaceutical sciences : a publication of the Canadian Society for Pharmaceutical Sciences, Societe canadienne des sciences pharmaceutiques, 18(5).

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