LC-MS/MS analysis was performed on an LC-30AD ultraperformance liquid chromatograph (Shimadzu, Japan) coupled with a QTRAP 6500 mass spectrometer (Applied Biosystems, Waltham, MA, USA). Chromatographic column (Waters ACQUITY BEH C18, 2.1 mm × 100 mm, 1.7 µm) was used for separating the FQs in honey samples. The mobile phases, consisting of A (methanol) and B (2 mmol/L ammonium formate containing 0.1% formic acid), were set at the flow rate of 0.3 mL/min. The elution gradient was as follows: 0–1.0 min (B: 95%), 1–3 min (B: 95–80%), 3–7.5 min (B: 80–5%), 7.5–10 min (B: 5%), 10–10.1 min (B: 5–95%), and 10.1–12.0 min (B: 95%). The injection volume of the sample extract was 5 µL. Source temperature was 500 °C and ion spray voltage was set at 4.5 kV; curtain gas, gas 1, and gas 2 pressures were set at 40 psi, 55 psi, and 50 psi, respectively. The MRM (multiple reaction monitoring) transition parameters are listed in the Supplementary Materials (Table S3) .
Lc 30ad
Manufactured by Shimadzu
Sourced in Japan, United States
The LC-30AD is a high-performance liquid chromatography (HPLC) pump from Shimadzu. It is designed to deliver precise and stable mobile phase flow for a variety of HPLC applications. The pump features a compact design, a maximum flow rate of 10 mL/min, and a pressure range up to 60 MPa.
Automatically generated - may contain errors
Lab products found in correlation
Sil 30ac, by Shimadzu (31 mentions)
Cbm 20a, by Shimadzu (15 mentions)
Cto 20ac, by Shimadzu (12 mentions)
Spd m20a, by Shimadzu (9 mentions)
Cto 30a, by Shimadzu (5 mentions)
Spd m30a, by Shimadzu (5 mentions)
Dgu 20a, by Shimadzu (5 mentions)
Dgu 20a5r, by Shimadzu (4 mentions)
Cto 20a, by Shimadzu (4 mentions)
Lcms 2020, by Shimadzu (4 mentions)
Nexera uhplc system, by Shimadzu (3 mentions)
Formic acid, by Thermo Fisher Scientific (3 mentions)
Uplc system, by Shimadzu (2 mentions)
Hplc system, by Shimadzu (2 mentions)
Lc 20ad, by Shimadzu (2 mentions)
Spd 20a, by Shimadzu (2 mentions)
Dgu 20a5, by Shimadzu (2 mentions)
Multiquant 3, by AB Sciex (2 mentions)
Cdd 10avp, by Shimadzu (2 mentions)
Kinetex c18 column, by Phenomenex (2 mentions)
81 protocols using lc 30ad
1
Quantitative Analysis of Fluoroquinolones in Honey
2
Quantitative Analysis of GOS Production
3
UHPLC Analysis of α-Dicarbonyl Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
α-DC analysis was conducted using an Ultrahigh Performance Liquid Chromatography apparatus (Shimadzu, Columbia, MD) consisting of two pumps LC-30AD, an SPD-M20A photo diode array detector (PDA), DGU–20A5 degasser, SIL–30AC autosampler (operated at 4°C) and CTO-20 AC column oven. The UHPLC system was controlled by a personal computer equipped with LabSolutions software operating in a Windows XP. The separation was achieved following the method described by Papetti et al. [18 (link)] using Ascentis Express ES-C18 column (150 × 4.6 mm, 2.7 μm particles; Sigma-Aldrich, St. Louis, MO) with a UHPLC pre-column filter (UltraShield Analytical Scientific Instruments, Richmond, CA) operating at 25.0 ± 0.5°C. The analytes were eluted with a flow rate of 0.3 mL/min using 0.1% formic acid in water (A) and methanol (B) as eluents. The 120 min gradient is described as follows: 0–5 min (90–85% A), 5–13 min (85–80% A), 13–40 min (80% A), 40–65 min (80–70% A), 65–90 min (70–50% A), 90–100 min (50–0% A), 100–105 min (0% A), and 105–110 min (0–90% A); the column was then re-equilibrated with the initial mobile phase for 10 min. The injection volume was 5 μL. The PDA detector was set at 314 and 335 nm to record the peaks, and UV spectra recorded from 215 to 420 nm. Before injection sample solutions were filtered with PVDF syringe filter (13 mm, 0.22 μm; Millipore Millex, Billerica, MA, USA).
4
Quantification of Acylcarnitines by Mass Spectrometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples were analyzed by a modified version of the flow analysis described by Millington and Stevens.21 (link) A Sciex QTRAP 6500 (Waverley, Victoria, Australia) tandem mass spectrometer coupled to a Shimadzu LC-30AD (Sydney, New South Wales, Australia) was used to flow-inject samples using a mobile phase of 50:50 methanol and water at 20 µL/min for 7 minutes. The mass spectrometer used a declustering potential of 150 V, ion spray voltage of 5500 V, and collision energy of 46 to conduct a precursor ion scan of m/z 99.1. The scan was conducted with the turbo spray source at 21°C and recorded as an accumulated spectrum from m/z 200 to 500. Gas settings were set at curtain gas (CUR) of 20, GS1 of 15, and GS2 of 15.
Areas under the peak for each acylcarnitine were calculated using a summation integration method in MultiQuant 3.0 (Waverley, Victoria, Australia). The areas for each acylcarnitine were analyzed as ratios to the internal standard, d3-palmitoylcarnitine, to generate relative concentrations. To account for interexperimental variability, the mean of the control samples was calculated and used to divide the relative concentration of acylcarnitine for each sample within the same experiment. This normalization was conducted for each experiment prior to pooling the data.
Areas under the peak for each acylcarnitine were calculated using a summation integration method in MultiQuant 3.0 (Waverley, Victoria, Australia). The areas for each acylcarnitine were analyzed as ratios to the internal standard, d3-palmitoylcarnitine, to generate relative concentrations. To account for interexperimental variability, the mean of the control samples was calculated and used to divide the relative concentration of acylcarnitine for each sample within the same experiment. This normalization was conducted for each experiment prior to pooling the data.
5
UHPLC-based Reversed-phase Separation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Reverse-phase separation was performed using a Nexera UHPLC (Shimadzu Corporation, Kyoto, Japan) composed of one degasser DGU-20A 5R, pump LC-30AD, autosampler SIL-30AC, and oven CTO-30A. Analytes were separated in a XSelect HSS T3 (150 mm × 2.1 mm I.D, particle size of 2.5 µm) analytical column at 40 °C, using 5 mM ammonium formate and 0.1% formic acid (v/v) in water as eluent A, and 5 mM ammonium formate and 0.1% formic acid (v/v) in methanol as eluent B. The gradient at a flow rate of 0.3 ml/min was as follows: 5% B from 0 to 1 min, then increased from 1 to 21 min to 90% B, constant from 21 to 25 min to 90% B, and decreased from 26 to 30 min to 5% B. The gradient was reduced for urine samples to 5% B from 0 to 1 min, then increased from 1 to 5 min to 90% B, constant from 5 to 9 min to 90% B, and decreased from 10 to 14 min to 5% B. Injection volume for mix 50 was 5 µl and for urine samples 10 µl.
6
Metabolite Profiling via QTRAP LC-MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The SCIEXSCIEX QTRAP 6500+ mass spectrometer and a Shimadzu LC30AD liquid chromatography system was used to analyze the supernatant. The Waters XBridge Amide (100 mm × 4.6 mm i.d., 3.5 μm) was used for LC separation. A 5μL sample was needed. The electrospray ionization mass spectra were acquired in positive ion mode (4850 V ion spray voltage) and negative ion mode (4500 V ion spray voltage), respectively. The multiple reaction monitoring (MRM) acquisition methods were used to collect MS information simultaneously. The heated capillary temperature was maintained at 475 °C. The curtain gas flow, nebulizer, and heater gas were set to 25, 33, and 33 arbitrary units, respectively.
7
Quantification of Neopterin in Silkworm Brains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ultra performance liquid chromatograph (UPLC) system (Shimadzu, Japan) used here consists of a SIL-30AC automatic sampler, an LC-30AD solvent delivery module, a CTO-30A column oven, a CBM-20A system controller, an RF-20A fluorescence detector and an LC solution workstation. The silkworm brains were removed immediately and placed into 1.5 ml centrifuge tubes containing 50 mM Tris-HCl (pH 7.5), 0.1 M KCl, 1 mM EDTA, 1 mM dithiothreitol and proteinase inhibitors by use of a JXFSTPRP-32 grinding miller (Shanghai Jingxin Industrial Development Co. Ltd, China) [37 (link)]. About 1 ml of the homogenizing buffer was added to each sample. After homogenizing, the mixtures were centrifuged at 12 000 rpm at 4°C for 20 min. Then, the supernatant was filtered using a 0.22 µm filter (Tianjin Jinteng Experiment Co. Ltd, China) and stored at −80°C. A T3 C18 column (1.8 µm, 2.1 internal diameter ×100 mm) (Acquity UPLC HSS) was used to separate the chemicals under the mobile phase isopropanol : methanol : acetic acid : H2O (0.5:0.5:0.5:98.5, v/v, pH 2.68) with a flow rate of 0.15 ml min−1. Neopterin was detected at λex/em 350/450 nm [37 (link),42 (link)].
8
HPLC Assay for Silmitasertib in Biological Samples
9
HPLC Separation of Analytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HPLC chromatographic separation was achieved through a Shimadzu apparatus (Kyoto, Japan) with system controller (CBM-20A), pump (LC-30AD), autoinjector (SIL-30AC), online degasser (DGU-20A), and column heater (CTO-20A). An Agilent Zorbax SB-C18 column with the diameter of 2.1 × 100 mm (3.5 μM, Agilent, USA) was applied to separate the analytes. The temperature of column was 40°C. Analytes were equivalently eluted with 30:70 (v/v) acetonitrile-water containing 5 mM ammonium acetate (pH 4) and the flow rate was 0.3 ml/min. The analysis time of all analytes was 1.8 min, and the injection volume was 10 μl.
10
Quantitative Analysis of Phenylalanine
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the cell culture medium, analysis of Phe was performed on LC-MS (QTRAP 5500; AB SCIEX) and HPLC (UFLC and LC-30AD; Shimadzu). All of the samples were injected into the liquid chromatography using a Waters HPLC BEH C18 column (2.1 × 50 mm, 1.7-μM particle size). The mobile phase (0.1% formic acid in water/methanol) was pumped at a flow rate of 0.5 mL/min. The mass spectrometry conditions were: ion detection method (multiple reaction mode), ion polarity (positive), declustering potential (120.0 V), entrance potential (10.0 V), collision energy (18.0 eV), collision cell exit potential (10.0 V), ion source (turbo spray), curtain gas (20.0 Psi), collision gas ( medium), ionspray voltage (5,500.0 V), temperature (500.0°C), ion source gas 1 (60.0 Psi), and ion source gas 2 (60.0 Psi).
For blood samples, the blood was collected at 4 p.m. according to previous reports,18 (link) and the whole blood was collected on the blood specimen collection card, dried under ambient conditions, and stored at 4 p.m. in plastic bags. The concentration of Phe in the blood was tested by dry blood spots, according to the protocol provided in the Phe assay kit.
For blood samples, the blood was collected at 4 p.m. according to previous reports,18 (link) and the whole blood was collected on the blood specimen collection card, dried under ambient conditions, and stored at 4 p.m. in plastic bags. The concentration of Phe in the blood was tested by dry blood spots, according to the protocol provided in the Phe assay kit.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!