Plasma concentrations of gliclazide were determined by a high-performance liquid chromatography-mass spectrometric (HPLC LC/MS) assay.13 (link) The method was referenced to the Bioanalytical Method Guidance as stated in the United States Food and Drug Administration. The calibration standards and quality control samples were prepared by spiking known amounts of gliclazide in drug-free plasma. The HPLC system consisted of a Waters Alliance 2690 separation module, Millenium chromatography management system and Waters 996 photodiode array detector (Waters, Milford, Mass).
Alliance 2690 separation module
Manufactured by Waters Corporation
Sourced in United States
The Alliance 2690 separation module is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative-scale separations. It features a modular design with a solvent delivery system, autosampler, and column compartment to enable flexible configuration for a variety of analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
X select hss ireland, by Waters Corporation (2 mentions)
2487 dual λ absorbance detector, by Waters Corporation (2 mentions)
Formic acid, by Thermo Fisher Scientific (2 mentions)
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
Reverse phase 5 m c 18 column, by Waters Corporation (2 mentions)
Model 2010a, by Shimadzu (2 mentions)
Synergi 4 m polar rp column, by Phenomenex (2 mentions)
Waters 996 photodiode array detector, by Waters Corporation (1 mentions)
Xbridge c18 analytical column, by Waters Corporation (1 mentions)
Amazon etd ion trap mass spectrometer, by Bruker (1 mentions)
Mightysil rp 18 column, by Kanto Chemical (1 mentions)
10 protocols using alliance 2690 separation module
1
Quantitative Analysis of Gliclazide in Plasma
2
HPLC Analysis of Hydroxy Fatty Acids
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All HPLC Analyses of (a)HETE and (a)HODE were carried out on a single Beckman
5 μm ultrasphere silica column (250 mm × 4.6 mm) using
isocratic normal phase conditions (1.2% IPA in hexanes containing
0.1% acetic acid).28 (link) Chiral HPLC analyses
of HETE and HODE methyl esters were performed on a Chiralpak AD column
(250 mm × 4.6 mm) produced by Chiral Technologies Inc., Exton,
PA. aHETE products have been eluted with 2% ethanol
in hexanes, whereas aHODE were eluted with 5% methanol
in hexanes. Direct infusion MS experiments were performed on ThermoFinnigan
TSQ Quantum triple quadrupole mass spectrometer, whereas all HPLC/MS
analyses were conducted on the same instrument coupled with a Surveyor
MS Pump and Surveyor Autosampler (for RP-HPLC) or with Waters Alliance
2690 Separation Module (NP-HPLC). Detailed information about solvent
gradients and MS settings applied during these analyses is given in
the appropriate protocols presented below. Unless stated otherwise,
all the HPLC separations were conducted with a solvent flow rate of
1 mL/min.
5 μm ultrasphere silica column (250 mm × 4.6 mm) using
isocratic normal phase conditions (1.2% IPA in hexanes containing
0.1% acetic acid).28 (link) Chiral HPLC analyses
of HETE and HODE methyl esters were performed on a Chiralpak AD column
(250 mm × 4.6 mm) produced by Chiral Technologies Inc., Exton,
PA. aHETE products have been eluted with 2% ethanol
in hexanes, whereas aHODE were eluted with 5% methanol
in hexanes. Direct infusion MS experiments were performed on ThermoFinnigan
TSQ Quantum triple quadrupole mass spectrometer, whereas all HPLC/MS
analyses were conducted on the same instrument coupled with a Surveyor
MS Pump and Surveyor Autosampler (for RP-HPLC) or with Waters Alliance
2690 Separation Module (NP-HPLC). Detailed information about solvent
gradients and MS settings applied during these analyses is given in
the appropriate protocols presented below. Unless stated otherwise,
all the HPLC separations were conducted with a solvent flow rate of
1 mL/min.
3
LC-IT-MS Analysis of Compounds
4
HPLC Analysis of Vitamins and Purines
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A Waters Alliance 2,690 separation module with UV detector was used for the chromatographic analysis (United States) coupled to a C18 stainless steel column with a 5 µm particle size (250 × 4.6 mm). For vitamin C, dopamine and noradrenaline analysis, methanol, HPLC grade water and 20 mM monobasic sodium phosphate (5:95, v/v) were used with isocratic elution at 280 nm at a column temperature of 35°C and an elution rate of 0.5 mL/min. For the analysis of adenosine, inosine and hypoxanthine, acetonitrile, HPLC grade water and 0.01 M monobasic sodium phosphate (5:95 v/v) was used with isocratic elution at 260 nm, with the column at room temperature and a flow rate of 1.0 mL/min (Ur Rehman et al., 2020 (link); Arif et al., 2022 (link)).
5
HPLC-FLD for Tetrodontoxin Analysis
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HPLC-FLD for TTX analysis was conducted as described previously [55 (link)]. An Alliance 2690 Separation Module (Waters) connected with the Waters 2487 fluorescence detector. A Mightysil RP-18 column (4.6 mm i.d × 250 mm, particle size 5 µm, Kanto Chemical Co., Inc) was used. The mobile phase for the TTX and TTX analogs was 2 mM heptanesulfonic acid in 10 mM ammonium phosphate buffer (pH 7.0) at a flow rate of 1 mL/min. The eluate was continuously mixed with 4 M NaOH and heated at 110 °C. The intensity of the fluorescence was measured at 505 nm with 384 nm excitation. The same TTX standard was used to compare the retention time of TTX and TTX analogs. The LOD of TTXs was 0.02 μg/g tissue (S/N = 3) and the LOQ of TTXs was 0.06 μg/g tissue; (S/N = 10).
6
HPLC Analysis of Vitamins and Purines
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A Waters Alliance 2,690 separation module with UV detector was used for the chromatographic analysis (United States) coupled to a C18 stainless steel column with a 5 µm particle size (250 × 4.6 mm). For vitamin C, dopamine and noradrenaline analysis, methanol, HPLC grade water and 20 mM monobasic sodium phosphate (5:95, v/v) were used with isocratic elution at 280 nm at a column temperature of 35°C and an elution rate of 0.5 mL/min. For the analysis of adenosine, inosine and hypoxanthine, acetonitrile, HPLC grade water and 0.01 M monobasic sodium phosphate (5:95 v/v) was used with isocratic elution at 260 nm, with the column at room temperature and a flow rate of 1.0 mL/min (Ur Rehman et al., 2020 (link); Arif et al., 2022 (link)).
7
Quantification of Neurotransmitters and Metabolites
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HPLC analysis was performed using a Waters Alliance 2690 separation module equipped with an auto-sampler, UV detector, and PDA (United States). A C18 column (250 × 4.6 mm, 5 µm particle size) (Waters X Select® HSS Ireland) was used, and the mobile phase employed for quantification of noradrenaline, serotonin, dopamine and vitamin C was composed of 20 mM monobasic sodium phosphate and methanol (95:5, v/v), the detection is performed at 280 nm, at a column temperature of 35°C and flow rate of 0.5 mL/min (Tokhi et al., 2023 (link)). The mobile phase used for quantification of adenosine, inosine, and hypoxanthine was composed of acetonitrile (5:95, v/v) and 0.01M monobasic sodium phosphate. The flow rate was 0.01 mL/min, at a column temperature of 35°C, and detection was performed at 260 nm using isocratic elution in phosphate buffer (Ur Rehman et al., 2020 (link)).
8
Chromatographic Analysis of Compounds
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Chromatographic analysis was performed utilizing a Waters Alliance 2690 separation module with PDA, UV detector, and autosampler (USA). A C18 stainless steel column (250× 4.6 mm) (Waters X Select® HSS Ireland) with a 5µm particle size was employed. The mobile phase comprised methanol and 20mM monobasic sodium phosphate (5:95, v/v); while detection was performed at 280 nm with isocratic elution. The elution rate was set at a flow rate of 0.5 mL/min while the column was maintained at a temperature of 35 ℃.29 (link),38 (link)
9
Quantifying PTX and BIBF in Nanoparticles
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NP were dissolved in acetonitrile and drug content was estimated through HPLC-UV for PTX and HPLC-MS for BIBF. PTX content in the NP was quantified using HPLC-UV (2690 Alliance separation module coupled with 2487 dual λ absorbance detector, Waters, Milford, MA). Reverse phase 5 µm C-18 column, 100 A°, 4.5 × 250 mm (Waters) was utilized in the assay and isocratic elution with a mobile phase of acetonitrile (Fisher Scientific): water (60:40, v/v) at a flow rate of 1 mL/min was used. The detection wavelength was set at 227 nm and the injection volume was 100 µL. BIBF content was determined using HPLC-Mass (Shimadzu Model 2010A liquid chromatograph and mass spectrometer, Shimadzu, Columbia, MD) using a LC-10AD VP Solvent Delivery system. Synergi 4 µm Polar-RP column, 80 A°, 2 × 150 mm (Phenomenex Inc, Torrance, California) was used. Isocratic elution was utilized with a mobile phase composed of water + 0.1% formic acid (Fisher Scientific): acetonitrile + 0.1% formic acid (50:50, v/v), at a flow rate of 0.2 mL/min. Electrospray ionization was used, m/z ratio of 540.5 was utilized, and 25 µL was injected.
Drug loadings and encapsulation efficiencies were calculated from the following formulas.
Drug loadings and encapsulation efficiencies were calculated from the following formulas.
10
Quantifying PTX and BIBF in Nanoparticles
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