Wps 3000tsl

Manufactured by Thermo Fisher Scientific

Sourced in United States, France

The WPS-3000TSL is a well plate sealer designed for sealing microplate wells. It can accommodate a variety of well plate sizes and sealing materials. The core function of this product is to provide a reliable and consistent method for sealing well plates.

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WPS-3000TSL autosampler (2 citations)

27 protocols using wps 3000tsl

Ultra-High-Performance Liquid Chromatography for Pharmaceutical Compound Analysis

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The sample analysis was performed by using a Dionex UltiMate 3000 UHPLC system (Thermo Scientific, San Jose, CA, USA), comprising a LPG-3400SD binary pump, WPS-3000TSL thermostat autosampler, TCC-3000SD thermostat column compartment and DAD-3000 diode array detector. The data were recorded and analyzed by Chromeleon™ 7.2 Chromatography Data System Software (Thermo Scientific, San Jose, CA, USA).
The chromatographic separation of MI, MCI, MP, EP, PP and BP was achieved through an ACCLAIM™ 120 C₈ analytical column with the dimensions 150 mm × 2.1 mm and 5 μm of particles size (Thermo Scientific, San Jose, USA). The optimal separation was obtained by using binary mobile phase: water (0.1% trifluoroacetic acid, pH 2.1, solvent A) and acetonitrile (solvent B) at a flow rate of 0.5 mL/min. The gradient mobile phase elution was 0–2 min (B, 12.5%), 2–4 min (B 20–30%), 4–16 min (B, 30–50%), 16–22 min (B, 50–100%), return to its equilibrium conditions and 22–30 min. The column temperature was kept at 35 °C, and the sample injection volume was 10 µL. The column was also washed with a mixture (50:50, v/v) of methanol and Milli-Q water solution, for five minutes, following the analysis of every ten samples. The optimal detection wavelength was performed in the UV range at 255 nm.

Ali H.M., Alsohaimi I.H., Khan M.R, & Azam M. (2020). Simultaneous Determination of Isothiazolinones and Parabens in Cosmetic Products Using Solid-Phase Extraction and Ultra-High Performance Liquid Chromatography/Diode Array Detector. Pharmaceuticals, 13(11), 412.

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Ultrafiltration Centrifugation for FH Entrapment Efficiency

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Drug EE% was determined
by the ultrafiltration centrifugation method.^{19 (link)} Briefly, 1 mL of FH-HEX was centrifuged in an Amicon centrifugal
tube (100 000 molecular weight cutoff (MWCO), Millipore) at
15 000 rpm for 15 min at 4 °C. The supernatant was then
collected, and the amount of free FH was quantified using a previously
validated HPLC (Dionex, Thermo Scientific), equipped with an LPG-3400SD
quaternary pump, a WPS-3000TSL autosampler, a variable-wavelength
detector (VWD-3000), and a reverse-phase C₁₈ column (Hypersil
ODS, 150 × 4.6 mm², 5 μm). The wavelength of
the UV detector was set at 226 nm. The samples were separated using
acetonitrile and deionized water containing 10 mM aqueous triethylamine
at a ratio of 55:45 v/v at a flow rate of 1 mL/min at 25 °C.
The coefficient of determination (R²)
of the FH calibration curve in PBS (pH 7.4) in the concentration range
of 1–100 μg/mL was 0.9994, and the respective limits
of detection (LOD) and quantification (LOQ) were 0.5 and 1 μg/mL.
The coefficients of variation (CV)% ranged from 2.1 to 4.9%, and the
accuracy for FH determination was 1.5–4.6% with a mean% drug
recovery of 97.5 ± 1.16%.
The adsorbed amount of FH on
the Amicon centrifugal tube membrane was determined by filtering an
FH solution of the same concentration, and then the amount of free
FH in the filtrate was quantified.^{56 (link)}EE% of FH was calculated using the following equation:

Abdel-Bar H.M., Khater S.E., Ghorab D.M, & Al-mahallawi A.M. (2020). Hexosomes as Efficient Platforms for Possible Fluoxetine Hydrochloride Repurposing with Improved Cytotoxicity against HepG2 Cells. ACS Omega, 5(41), 26697-26709.

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Monoamine Neurotransmitter Extraction and Analysis

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Dissected colon and ileal segments were placed in a buffer comprising 0.2 M perchloric acid, 0.05% Na₂EDTA, 0.1% Na₂S₂O₅, and isoproterenol (internal standard) and stored at −80°C until lysate preparation. To extract monoamine neurotransmitters, the samples were homogenized using a bullet blender and 0.2-mm diameter stainless steel beads. Lysates were centrifuged and 300 μL of lysate was added to 30 mg alumina. Lysate pH was quickly increased to 8.6 using 1 M Tris buffer. Lysates were mixed in a rotating shaker at 4°C for 15 min and then centrifuged at 10,000 x g. Supernatants were aspirated and the monoamine neurotransmitters were eluted from the alumina with 0.2 M perchloric acid. Monoamine lysates were placed in a refrigerated automatic sampler (model WPS-3000TSL) until being separated isocratically by a reversed-phase C18 column with a flow rate of 0.6 mL/min using a Dionex Ultimate 3000 HPLC system (pump ISO-3100SD, Thermo Scientific, Chelmsford, MA). Electrochemical detection was achieved using an ESA CoulArray model 5600A (Thermo Fisher) coupled with an analytical cell (microdialysis cell 5014B) and a guard cell (model 5020) with cell potentials set at −350, 0, 150, and 220 mV. Data acquisition and analysis were performed using Chromeleon 7 and ESA CoulArray 3.10 HPLC software and quantified data were normalized to wet tissue weight.

Ghaisas S., Langley M.R., Palanisamy B.N., Dutta S., Narayanaswamy K., Plummer P.J., Sarkar S., Ay M., Jin H., Anantharam V., Kanthasamy A, & Kanthasamy A.G. (2019). MitoPark Transgenic Mouse Model Recapitulates the Gastrointestinal Dysfunction and Gut-Microbiome Changes of Parkinson’s Disease. Neurotoxicology, 75, 186-199.

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Analytical Methods for Compound Characterization

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LC analysis was performed in a Dionex Ultimate 3000 liquid chromatograph with an autosampler (WPS-3000TSL), a pump (LPG-3400SD), a column thermostat (TCC-3000SD), and a diode detector (DAD-3000) (Thermo-Fisher Scientific, Sunnyvale, CA, USA). The system was connected with Chromeleon 7 software.
Spectrophotometric measurements were conducted using a Multiskan GO Spectrophotometer (Thermo-Fisher Scientific). The %Inhibition was calculated from the formula

% inhibition = (\frac{A_{Control} - A_{Sample}}{A_{Control}}) \times 100

MS spectra in the negative mode were recorded on an ESI-qTOF Compact mass spectrometer (Bruker, Bremen, Germany). The mass spectrometer was re-calibrated for every run [31 (link)]. ¹H-NMR, ¹³C-NMR, HSQC, HMBC, and COSY experiments were recorded on Avance 300 MHz spectrometer (Bruker) in DMSO-d6 and calibrated using the residual solvent peak. The data were processed with MestReNova 12 software (Mestrelab Research, Santiago de Compostela, A Coruña, Spain).

Starzec A., Włodarczyk M., Kunachowicz D., Dryś A., Kepinska M, & Fecka I. (2023). Polyphenol Profile of Cistus × incanus L. and Its Relevance to Antioxidant Effect and α-Glucosidase Inhibition. Antioxidants, 12(3), 553.

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HPLC Analysis of Monoamine Neurotransmitters

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Striatum and olfactory bulb samples were prepared and processed for HPLC as described previously (Gordon et al., 2016a ). Briefly, dissected brain regions were placed in a buffer containing 0.2 M perchloric acid, 0.05% Na2EDTA, 0.1% Na2S2O5, and isoproterenol (internal standard) to extract monoamine neurotransmitters. Monoamine lysates were placed in a refrigerated automatic sampler (model WPS-3000TSL) until being separated isocratically by a reversed-phase C18 column with a flow rate of 0.6 ml/min using a Dionex Ultimate 3000 HPLC system (pump ISO-3100SD, Thermo Scientific, Bannockburn, IL). Electrochemical detection was achieved using a CoulArray model 5600A coupled with an analytical cell (microdialysis cell 5014B) and a guard cell (model 5020). Data acquisition and analysis were performed using Chromeleon 7 and ESA CoulArray 3.10 HPLC Software and quantified data were normalized to wet tissue weight.

Langley M.R., Ghaisas S., Ay M., Luo J., Palanisamy B.N., Jin H., Anantharam V., Kanthasamy A, & Kanthasamy A.G. (2017). Manganese exposure exacerbates progressive motor deficits and neurodegeneration in the MitoPark mouse model of Parkinson's disease: Relevance to gene and environment interactions in metal neurotoxicity. Neurotoxicology, 64, 240-255.

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Characterization of Protein Samples

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At‐line MALS and fluorescence measurements were performed using an Ultimate 3000 system (Thermo Fisher, Waltham, MA, USA). A sample of all collected fractions during the purification runs was directly injected into the detectors bypassing the column. The HPLC system was equipped with a LPG‐3400SD quaternary pump, WPS‐3000TSL analytical autosampler, DAD 3000 UV‐detector and FLD 3100 fluorescence detector (Thermo Fisher, Waltham, MA, USA). Additionally, the system was connected to a multi‐angle light scattering detector DAWN HELEOS 18‐angle and a differential refractive index detector Optilab rEX (both Wyatt, Santa Barbara, CA, USA). Chromeleon 7 (Thermo Fisher Scientific, Waltham, MA, USA) and Astra 5.3.4 Wyatt, (Santa Barbara, CA, USA) software were used for method programming, system control and data acquisition. GFP fluorescence was monitored with an excitation wavelength of 480 nm and emission of 505 nm. Analysis time was 3 min/sample. Light scattering intensity was accessed by calculating the peak area of the light scattering signal obtained with the 90° angle.

Pereira Aguilar P., González‐Domínguez I., Schneider T.A., Gòdia F., Cervera L, & Jungbauer A. (2019). At‐line multi‐angle light scattering detector for faster process development in enveloped virus‐like particle purification. Journal of Separation Science, 42(16), 2640-2649.

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Quantitative LC-MS/MS Bioanalysis Protocol

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A TSQ Quantum Ultra mass spectrometric detector with electrospray ionization (ESI) interface (Thermo Scientific, USA) was coupled with a Dionex Ultimate 3000 ultra-performance liquid chromatography system consisting of an LPG-3400SDN pump, a WPS-3000TSL autosampler, and a TCC-3000RS column compartment. The separation of analytes and IS was achieved by using a Hypersil GOLD C18 column (50 mm × 2.1 mm, 3 µm; Thermo Scientific, USA) that was maintained at 25°C. The water (containing 0.1% formic acid and solvent A) and acetonitrile (solvent B) was used as mobile phase with gradient elution. The gradient was as follows: 0 min 5% B, 0.5 min 5% B, 2.0 min 80% B, 2.1 min 5% B, and 3.0 min 5% B, and then stopped. The flow rate was set at 0.4 mL/min. The select reaction monitoring (SRM) conditions (source parameters) were defined as follows: spray voltage 3.5 kV, vaporizer temperature 250°C, and capillary temperature 300°C. The optimized mass spectrometric parameters and mass spectra for all the analytes are shown in Table 1 and Figure 1, respectively. All the operations, acquiring and analysis of data, were controlled using Xcalibur software (Thermo Scientific, USA).

Cui W., Liu Q., Xiong S, & Qiao L. (2018). LC-MS/MS Method for Simultaneous Quantification of Dexmedetomidine, Dezocine, and Midazolam in Rat Plasma and Its Application to Their Pharmacokinetic Study. Journal of Analytical Methods in Chemistry, 2018, 3184759.

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HPLC-ECD Analysis of Neurotransmitters

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The assay was validated in a previous method for HPLC-ECD detection of tyrosine and tryptophan-based metabolites (Langley et al., 2017 (link)). The brain tissue lysates were loaded in the auto-sampler (Thermo Scientific WPS-3000TSL). The analytes were separated isocratically with mobile phase (MDTM, Thermo-Scientific) by reversed-phase C-18 Column (ZORBAX Eclipse Plus 4.6 × 100 mm, Agilent) at 0.6 ml/min for 21 min (pump, ISO-3100 SD, Thermo Scientific). Detection of metabolites DA, NE, DOPAC, HVA, 5-HIAA, and 5-HT were performed by ECD (CoulArray 5600A) using the following potentials: Channel 1: 350 mV (ESA Guard cell, 5020) Channel 3: −150 mV and CH4: +220 mV (ESA Microdialysis cell, 5014B). Method development and Data integration and analysis were performed using Chromeleon (v. 7.1.3) and ESA CoulArray Software (v. 3.10).

Massey N., Shrestha D., Bhat S.M., Padhi P., Wang C., Karriker L.A., Smith J.D., Kanthasamy A.G, & Charavaryamath C. (2022). Mitoapocynin Attenuates Organic Dust Exposure-Induced Neuroinflammation and Sensory-Motor Deficits in a Mouse Model. Frontiers in Cellular Neuroscience, 16, 817046.

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Quantifying Dopamine and Metabolites

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Brain STR were micro-dissected, preserved in antioxidant mixture containing 0.2 M perchloric acid, 0.1% NA2S2O5, 0.05% NaEDTA, and isoproterenol as an internal standard as described previously and immediately stored in dry ice [46 (link)]. The samples were further processed for HPLC-ECD with pump (Thermo-Scientific ISO-3100SD-BM) and autosampler (Thermo-Scientific WPS-3000 TSL) using a CoulArray 5600A-ECD detection system operated at 25 °C. Specifically, 20 µL samples were injected onto an Agilent Eclipse Plus (3.5 µm, 100A) C18 HPLC column (150 × 4.60 mm). Then, analytes were eluted with MD-TM MP mobile phase (Cat. no. 701332, Thermo Fisher Scientific) under isocratic conditions at 0.6 mL/min for 21 min. Dopamine (DA) and its major metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), were quantified using a standard curve. All sample concentrations were corrected using isoproterenol (ISO) as an internal standard correction factor and expressed as ng neurotransmitter/mg wet tissue weight.

Schlichtmann B.W., Palanisamy B.N., Malovic E., Nethi S.K., Padhi P., Hepker M., Wurtz J., John M., Ban B., Anantharam V., Kanthasamy A.G., Narasimhan B, & Mallapragada S.K. (2023). Aggregation-Inhibiting scFv-Based Therapies Protect Mice against AAV1/2-Induced A53T-α-Synuclein Overexpression. Biomolecules, 13(8), 1203.

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HPLC Analysis of Molecular Compounds

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The HPLC Dionex Ultimate 3000 system (Thermo Scientific, Villebon sur Yvette, France) contained an integrated solvent and degasser SRD-3200, an analytical pump HPG-3200SD, a thermostated autosampler WPS-3000TSL, a thermostated column compartment TCC-3000SD, and a diode array detector MWD-3000. Data acquisition (e.g., peak time and area) was carried out using in line Chromeleon software (v6.80 SP2) (Thermo Scientific). The eluent was monitored at 214 nm. Chromatographic separation was achieved at 25°C using a reverse phase Nova-Pak C18 column (60 Å, 4 μm, 4.6 mm × 150 mm, Waters, Guyancourt, France). The mobile phase (0.01 M phosphate buffer pH 3: methanol; 40 : 60 v/v) was pumped at a flow rate of 1.0 mL/min. The injection volume was set at 25 μL.

Ajemni M., Balde I.B., Kabiche S., Carret S., Fontan J.E., Cisternino S, & Schlatter J. (2015). Stability-Indicating Assay for the Determination of Pentobarbital Sodium in Liquid Formulations. International Journal of Analytical Chemistry, 2015, 697937.

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