A Shimadzu HPLC-UV system with a pump (LC-20AT), an autosampler (SIL-20AC), a column oven (CTO-20A), and an ultraviolet detector (SPD-20A) was used in this study (Shimadzu Co., Kyoto, Japan). A Kinetex C18 column (250 × 4.6 mm, 5 μm, 100 Å; Phenomenex, Torrance, CA, USA) protected by a C18 guard column (SecurityGuard HPLC Cartridge System, Phenomenex) was used for chromatographic separation at 40 °C. The mobile phase for the HPLC system consisted of phosphate buffer (pH 5.0; 10 mM) (solvent A) and ACN (solvent B) at a flow rate of 1 mL/min. The gradient elution protocol was as follows (solvent A/solvent B (v/v)): ramped from 71:29 (v/v) to 59:41 (v/v) during 10 min; back to 71:29 (v/v) for 10 min. The wavelength for SBN and IS was set as 220 nm. The total run time and injection volume were 20 min and 20 μL, respectively. All solvents used for sample preparation and HPLC analysis were degassed by sonication under vacuum and in-line electronic vacuum degasser modules.
Hplc uv system
Manufactured by Shimadzu
Sourced in Japan
The HPLC-UV system by Shimadzu is a high-performance liquid chromatography instrument equipped with a UV detector. It is designed to separate, identify, and quantify various chemical compounds in a liquid mixture through the process of liquid chromatography. The system utilizes a high-pressure pump to move the sample mixture through a column packed with a stationary phase, and a UV detector to measure the absorbance of the compounds as they elute from the column.
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Lab products found in correlation
Securityguard hplc cartridge system, by Phenomenex (1 mentions)
Spd 20a, by Shimadzu (1 mentions)
Lc 20at, by Shimadzu (1 mentions)
Kinetex c18 column, by Phenomenex (1 mentions)
Cto 20a, by Shimadzu (1 mentions)
Sil 20ac, by Shimadzu (1 mentions)
Spd 20a detector, by Shimadzu (1 mentions)
Lc 20at pump, by Shimadzu (1 mentions)
Cbm 20a controller, by Shimadzu (1 mentions)
5 protocols using hplc uv system
1
HPLC-UV Analysis of Pharmaceuticals
2
HPLC-UV Analysis of Organic Compounds
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A Shimadzu HPLC-UV system (Shimadzu Corporation, Kyoto, Japan) was used. The HPLC conditions were as follows: Column, YMC Pack ODS-AM, 250 mm × 6 mm I.D. column (YMC CO., LTD, Kyoto, Japan); eluent, a linear gradient of acetonitrile (0–50%B) in 0.1% (v/v) TFA for 50 min. Column temperature was 40 °C and the flow rate and detection wavelength were 1.0 mL/min and 270 nm, respectively. For fractionation, we injected 200 μL, and for the other purpose, we injected 20 μL.
3
Platelet Aggregation Analysis Using QX-100
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QX-100 platelet aggregation analyzer was obtained from Shanghai Fudan University Instrument factory (Shanghai, China). Drug analysis was performed with a Shimadzu HPLC-UV system (Tokyo, Japan), comprising a LC-10AD pump, an UV-visible detector, and a CTO-10A column oven. Milli-Q pure water instruments were from Millipore Co. Ltd. (Hong Kong, China).
4
Quantitative HPLC Analysis of NFOH in Rat Plasma
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The Ethics Committee approved the animal experiment protocol (protocol n°1155/2019). NLC-NFOH (2.8 mg/kg) was administered to one healthy male Wistar rat (341g) by gavage. After 5 h blood from the carotid vein was collected. A blood sample was collected to heparin tubes (10 µl, 500 IU/mL). Plasma samples were obtained immediately by centrifuging at 8000 rpm for 10 min, and stored at -20°C until further analysis by HPLC [24] (link). Plasma samples (200 µL) were mixed with acetonitrile (200 µL), to precipitate proteins, then centrifuged at 10000 rpm for 10 min. The supernatant was transferred to a vial insert and analyzed in HPLC.
The quantification of NFOH in plasma was performed according to Monteiro et al [25] (link).
The Shimadzu HPLC-UV system consisted of a CBM-20A controller, LC-20AT-pump, SPD-20A
detector, and SIL-20AC sampler LC Solution software (version 1.25SP4) was applied for data collecting and processing. The mobile phase consisted of acetonitrile: water (20:80) at a flow rate of 1.2 mL/min and the UV detector at 265 nm. The volume of injection was 20 μl. The method was linear (r= 0.9997) in the concentration range of 10 -0.1 µg/mL.
The quantification of NFOH in plasma was performed according to Monteiro et al [25] (link).
The Shimadzu HPLC-UV system consisted of a CBM-20A controller, LC-20AT-pump, SPD-20A
detector, and SIL-20AC sampler LC Solution software (version 1.25SP4) was applied for data collecting and processing. The mobile phase consisted of acetonitrile: water (20:80) at a flow rate of 1.2 mL/min and the UV detector at 265 nm. The volume of injection was 20 μl. The method was linear (r= 0.9997) in the concentration range of 10 -0.1 µg/mL.
5
HPLC-UV Analysis of Phytoestrogens
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The chromatographic analyses were performed according to Zimila et al. 10 on a Shimadzu HPLC-UV system consisting of a controller SCL-10AVP, degasser CTO-20A, pump LC-20AT and detector SPD-20A. The separation was carried out at 40°C on an ODS4 C18 column (5 μm, 4.6 mm x 150 mm, GLScience Inc., Tokyo), protected by a pre-column with the same composition (4.0 mm × 10 mm). A mobile phase consisting of a mixture of acetonitrile:water (77:23) was eluated in an isocratic mode at a flow rate of 1.0 mL min -1 . The wavelength was set at 282 nm and the run time was 20 min. The chromatographic peaks of PEs were in the range of 10 to 15.5 min, and the methanol solutions of DHPB at 2.5, 5.0, 10.0, 25.0 and 50.0 ng μL -1 were used to obtain the calibration curve.
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