The stored reaction mixture solutions or fermentation extracts were subjected to HPLC analysis (Dionex UltiMate 3000 SD HPLC system, Thermo Scientific, MA, USA). The separation was achieved on a C18 column (SilGreen ODS column [φ 4.6 × 250 mm, S-5 μM], Greenherbs, Beijing, China) with a flow rate of 1 ml/min at 40°C. A linear gradient elution was performed with mobile phases containing acetonitrile (A) and H2O (B). For detection of the product of K19G: 0 min: 40% A in 60% B; 0–10 min: linear gradient increase to 80% A in 20% B; 10–15 min: 100% A; 15–16 min: 40% A in 60% B; 16–20 min: 40% A in 60% B. For detection of the product of S19G, Sm: 0 min: 20% A in 80% B; 0–10 min: gradient increased to 60% A in 40% B; 10–15 min: 100% A; 15–16 min: 20% A in 80% B; 16–20 min: 20% A in 80% B. For detection of the product of Sb, Rub: 0 min: 15% A in 85% B; 0–10 min: line gradient increase to 55% A in 45% B; 10–15 min: 100% A; 15–16 min: 15% A in 85% B; 16–20 min: 15% A in 85% B. For detection of the product of stevioside, RedA, RedD: 0 min: 5% A in 95% B; 0–10 min: line gradient increase to 45% A in 55% B; 10–15 min: 100% A; 15–16 min: 5% A in 95% B; 16–20 min: 5% A in 95% B.
Ultimate 3000 sd hplc system
Manufactured by Thermo Fisher Scientific
Sourced in United States, Netherlands
The UltiMate 3000 SD HPLC system is a high-performance liquid chromatography instrument designed for versatile analytical applications. It features a modular design, advanced control software, and precision components to deliver reliable and reproducible separation of a wide range of analytes.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using ultimate 3000 sd hplc system
1
HPLC Analysis of Metabolite Products
2
UPLC Separation of Fluorescently Labeled Glycans
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Fluorescently labeled glycans were separated
on an Acquity UPLC glycan BEH amide column (2.1 × 100 mm, 1.7
μm, Waters Chromatography BV, Etten-Leur, Netherlands), using
an UltiMate 3000 SD HPLC system (Thermo Fisher Scientific, Waltham,
MA, U.S.A.) equipped with a Jasco FP-920 fluorescence detector (λex, 330 nm; λem, 420 nm; Jasco, Inc., Easton,
MD, U.S.A.). An injection volume of 3 μL was used. Ternary gradients
were run using Milli-Q water, acetonitrile, and a buffer solution
consisting of 250 mM formic acid in Milli-Q water, adjusted to pH
3.0 using ammonia. A constant 20% of the buffer was maintained throughout
the run. Elution was performed by a slow sloping gradient of 22–40%
Milli-Q water (total concentration, including buffer) from 0 to 67.5
min. The remaining percentage of the solvent composition comprised
of acetonitrile. After completion of the gradient, final gradient
conditions were maintained for 9 min and the column reconditioned
back to initial conditions for 13 min.
on an Acquity UPLC glycan BEH amide column (2.1 × 100 mm, 1.7
μm, Waters Chromatography BV, Etten-Leur, Netherlands), using
an UltiMate 3000 SD HPLC system (Thermo Fisher Scientific, Waltham,
MA, U.S.A.) equipped with a Jasco FP-920 fluorescence detector (λex, 330 nm; λem, 420 nm; Jasco, Inc., Easton,
MD, U.S.A.). An injection volume of 3 μL was used. Ternary gradients
were run using Milli-Q water, acetonitrile, and a buffer solution
consisting of 250 mM formic acid in Milli-Q water, adjusted to pH
3.0 using ammonia. A constant 20% of the buffer was maintained throughout
the run. Elution was performed by a slow sloping gradient of 22–40%
Milli-Q water (total concentration, including buffer) from 0 to 67.5
min. The remaining percentage of the solvent composition comprised
of acetonitrile. After completion of the gradient, final gradient
conditions were maintained for 9 min and the column reconditioned
back to initial conditions for 13 min.
3
Plasma Catechols Quantification by HPLC-ED
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Plasma samples were thawed on ice and a spatula-tip (~5mg) of activated alumina powder (199966, Sigma) was added to each well of a 96-well AcroPrep filter plate with 0•2 µM wwPTFE membrane (514-1096, VWR). A100 µL of plasma sample, 1 µM DHBA (3,4-dihydroxybenzylamine hydrobromide, 858781, Sigma) as an internal standard, and 800 µL of TE buffer (2•5% EDTA; 1•5 M Tris/HCl pH 8•6) were added sequentially to the wells. Liquid was removed using a 96-well plate vacuum manifold and the alumina were washed twice with 800 µl of H2O. Catechols were eluted using 0•7% HClO4, which was incubated for 30 min at RT. Samples were injected in a HPLC-ED system (Ultimate 3000 SD HPLC system coupled to Ultimate 3000 ECD-3000RS electrochemical detector with a glassy carbon working electrode (DC amperometry at 800 mV), Thermo Scientific). Samples were analysed on a C18 column (Kinetex 5 μM, C18 100 Å, 250 × 4•6 mm, Phenomenex, Utrecht, The Netherlands) using a gradient of water/methanol with 0•1% formic acid (0-3 min, 99% H2O; 3-7 min, 99-30% H2O; 7-10 min 30-5% H2O; 10-11 min 5% H2O; 11-18 min, 99% H2O). Data recording and analysis were performed using Chromeleon software (version 6•8 SR13). Potential intake differences of levodopa were corrected by using carbidopa as an internal standard.
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