DNA was isolated from tissue specimens or cell pellets as described11 (link). To quantify acetaldehyde-derived DNA damage, we measured the N2-ethylidene-dG level in the oesophagus of mice and cultured cells as described37 (link). Briefly, NaBH3CN (100 mM) (Sigma-Aldrich), a reducing reagent, was added to DNA samples. This converts N2-ethylidene-dG to stable N2-Et-dG. As the endogenous N2-Et-dG level in tissues is extremely low, the N2-Et-dG level that is converted from N2-ethylidene-dG indicates the endogenous N2-ethylidene-dG level10 (link). The DNA adduct standard, N2-Et-dG, and its stable isotope, [U-15N5]-labeled N2-Et-dG, were synthesized as described11 (link). DNA samples were digested as described11 (link), and subjected to liquid chromatography tandem mass spectrometry (LC/MS/MS). LC/MS/MS analyses were performed using a Shimadzu LC system (Shimadzu Corp., Kyoto, Japan) interfaced with a Quattro Ultimo triple-stage quadrupole mass spectrometer or an ACQUITY UPLC H-Class system interfaced with a XEVO-TSQ triple-stage quadrupole mass spectrometer (Waters Corp., Milford, MA, USA) as reported11 (link). Shim-pack XR-ODS columns (3.0 × 75 mm, 2.2 μm; Shimadzu Corp.) or ACQUITY UPLC BEH C18 columns (2.1 × 100 mm, 1.7 μm; Waters Corp.) were used to separate the samples.
Lc system
Manufactured by Shimadzu
Sourced in Japan, Germany
The Shimadzu LC system is a high-performance liquid chromatography (HPLC) instrument designed for the separation, identification, and quantification of a wide range of chemical compounds. It is a versatile analytical tool used in various industries, including pharmaceuticals, environmental analysis, food and beverage, and chemical research.
Automatically generated - may contain errors
Lab products found in correlation
Sil 20a ht, by Shimadzu (3 mentions)
Atlantis c18 column, by Waters Corporation (3 mentions)
8030 triple quadrupole mass spectrometer, by Shimadzu (3 mentions)
Nabh3cn, by Merck Group (2 mentions)
Acquity uplc beh c18 column, by Waters Corporation (2 mentions)
Acquity uplc h class system, by Waters Corporation (2 mentions)
Quattro ultimo triple stage quadrupole mass spectrometer, by Waters Corporation (2 mentions)
Xevo tsq triple stage quadrupole mass spectrometer, by Waters Corporation (2 mentions)
Shim pack xr ods column, by Shimadzu (2 mentions)
Lc 20ad pump, by Shimadzu (2 mentions)
Cto 10as vp, by Shimadzu (2 mentions)
Dgu 20a5r, by Shimadzu (2 mentions)
Luna c18 column, by Phenomenex (2 mentions)
Sil 20a, by Shimadzu (2 mentions)
Lc 20at pump, by Shimadzu (2 mentions)
Analyst tf 1, by AB Sciex (2 mentions)
Tripletof 5600 mass spectrometer, by AB Sciex (2 mentions)
Zorbax sb c8 reversed phase column, by Agilent Technologies (2 mentions)
Lc 20ad, by Shimadzu (2 mentions)
Sil 20ac autosampler, by Shimadzu (2 mentions)
68 protocols using lc system
1
Quantification of Acetaldehyde-Derived DNA Damage
2
Quantifying Esophageal N2-Ethylidene-dG Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Esophageal N2-ethylidene-dG level was quantified as described previously (44 (link)). Briefly, DNA was isolated from esophageal tissue specimens. NaBH3CN (100 mM; 156159, Sigma–Aldrich, St. Louis, MO), a reducing reagent, was added to DNA samples. This converts N2-ethylidene-dG to stable N2-Et-dG. As the endogenous N2-Et-dG level in tissues is extremely low, the N2-Et-dG level that is converted from N2-ethylidene-dG indicates the endogenous N2-ethylidene-dG level (45 (link)). The DNA adduct standard, N2-Et-dG and its stable isotope, [U-15N5]-labeled N2-Et-dG, were synthesized as described previously (46 (link)). DNA samples were digested as described previously (46 (link)) and subjected to liquid chromatography-tandem mass spectrometry (LC/MS/MS). LC/MS/MS analyses were performed using a Shimadzu LC system (Shimadzu Corp., Kyoto, Japan) interfaced with a Quattro Ultimo triple-stage quadrupole mass spectrometer or an ACQUITY UPLC H-Class system interfaced with a XEVO-TSQ triple-stage quadrupole mass spectrometer (Waters Corp., Milford, MA), as reported previously (46 (link)). Shim-pack XR-ODS columns (3.0 × 75 mm, 2.2 μm; Shimadzu Corp.) or ACQUITY UPLC BEH C18 columns (2.1 × 100 mm, 1.7 μm; Waters Corp.) were used to separate the samples.
3
Quantification of Fluoxetine by HPLC-FLD
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The quantification of FLX was performed by HPLC-FLD using a Shimadzu LC system (Shimadzu Corporation, Kyoto, Japan) equipped with a LC-20AD pump, a DGU-20A 5R degasser, a CTO-10AS VP column oven, a SIL-20A HT automatic injector, and a RF-20A-XS fluorescence detector. The chromatographic separation was achieved using a Luna C18 column (150 × 4.6 mm, 5 μm particle size) (Phenomenex, Torrance, CA, USA), using the method described by Silva et al. [20 (link)]. The identification of the analyte was based on its retention time compared with a standard solution, and the quantification was performed using the external calibration curve method. A linear relationship between peak area and the FLX concentration was established in the range of 1–2000 μg·L−1, according to Equation (S3) (Supplementary Materials) . The limit of detection (LOD) and limit of quantification (LOQ) were determined on the basis of the signal-to-noise ratio using the analytical response of 3 and 10 times the background noise, respectively. The determined LOD and LOQ for FLX were 10.8 and 35.9 μg·L−1, respectively.
4
Amino Acid Composition Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hydrolysis of total protein and the analysis of amino acid composition was performed in duplicate according to Vieira et al. (2018) [17 (link)] methodology. The liquid chromatograph consisted of a Shimadzu LC system (Shimadzu Corporation, Kyoto, Japan) equipped with an LC-20AD pump, DGU-20AS degasser and photodiode array SPD-M20A (PAD), and fluorescence RF-10AXL (FLD) detectors. The relative amino acid composition was expressed as mg/g protein.
5
HPLC Analysis of Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HPLC method was adapted from Ang et al. The HPLC analysis was performed using a Shimadzu-LC system (Shimadzu, Nakagyo-ku, Kyoto, Japan) equipped with a CBM-20A controller, LC-20AT pump, DGU-20A5 prominence degasser, SIL-20A auto-sampler, SPD-20AV detector and a CTO-10ASvp column oven. Chromatographic separations were achieved using the Thermo Hypersil Gold column (250 × 4.6 mm I.D.; 5 µm) guarded with C-18 guard column (Zorbax Eclipse Plus, Agilent, Santa Clara, CA, United States). The mobile phase which consists of acetonitrile and 2% acetic acid at the volume ratio of 40:60 (% v/v), was used with an isocratic elution at the flow rate of 1.3 mL/min. The column temperature was set at a standard of 35 °C with UV detection at 370 nm. The injection volume for each solution was 20 µL with a run time of 18.5 min. Data was acquired and processed using the LC-Solution Software (LC2010, Shimadzu, Nakagyo-ku, Kyoto, Japan). Solvents and distilled water were filtered via a 0.45 µm nylon membrane prior to being used in the HPLC method.
6
HPLC Quantification of Imiquimod
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Quantification of IMQ was performed based on a reported validated HPLC method.31 (link) A Shimadzu® LC system (Shimadzu Corporation, Kyoto, Japan) was used throughout this study. The machine was equipped with a pump (LC-20ADXR), an autosampler (SIL-20A HT), a degasser (DGU-20A3) and a photodiode array detector (SPD-M20A). PhenoSphere-Next™ C18 column (250 × 4.6 mm, 5 µm) was used, while the mobile phase consists of 0.1 M sodium acetate buffer adjusted to pH 4 and acetonitrile at a ratio of 60:40 (v/v). The volume of injection was 10 µL and a flow rate of 1.0 mL/min was applied. The total runtime was 5.5 minutes at which the peak was detected at 4.5 minutes using the wavelength of 244 nm.
7
Quantifying Phytohormones via HPLC-ESI-MS/MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A triple quadrupole mass spectrometer coupled with an electrospray ionization and high-performance liquid chromatography (HPLC‒ESI‒MS/MS) system (Shimadzu LC system, pump model: LC-10ADvp; oven model: CTO-10Avp; system controller model: SCL-10Avp; ABI 4000; Applied Biosystems)71 (link) was used to quantify phytohormones at Nanjing Ruiyuan Biotechnology Co., Ltd. (Nanjing, China).
8
GBP Quantification Using LC-MS/MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plasma concentrations of GBP were analyzed using a validated LC-MS/MS method. LC system (Shimadzu®, Japan) coupled with triple quadrupole MS/MS detector (API 4000®, Canada) was used. The chromatographic separation was carried out on Gemini C18 column (2.0 × 150 mm i.d., 5 μm; Phonomenex Inc. Torrance, CA, USA). The mobile phase consisted of 0.5% acetic acid:acetonitrile [10:90] (v/v).The flow rate was set as 0.5 ml/min. The analysis was operated at the MRM (multiple reaction monitoring) mode, and its MS parameters are shown in Table 2 (Park et al., 2007 (link)).
9
Quantitative Analysis of Cellular Glutathione
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
1x 106 cells were plated in 100 mm plates. The following day, cells were treated accordingly for 24h. Following treatment, media was removed, and plates were washed 1X and scraped into ice-cold PBS. Cells were pelleted via centrifugation at 1,000×g (5 min, 4 °C), and resuspended in 20 % 5-sulfosalcylic acid ((w/v) 150 μL) containing 1.25 nmol of GSH-(glycine-13C2,15N, internal standard). Samples were briefly sonicated and then centrifuged (14,000×g, 10 min). Supernatant (12 μL) was then chromatographed using a Shimadzu LC system equipped with a 150 × 3 mm, 3 μm particle diameter Atlantis C18 column (Waters, Milford, MA) at a flow rate of 0.400 mL/min. Solvent A (10 mM heptafluorobutyric acid (HFBA) in H2O) was held at 95 % for 1 min, and lowered to 90 % A over the next 9 min. Then, a linear gradient to 98 % B (10 mM HFBA in ACN) was applied over the next 10 min. The column was held at 98 % B for 4.5 min and then equilibrated to 95 % A for 0.5 min. The needle was washed prior to each injection with a buffer consisting of 25 mM NH4OAc in MeOH for 2.5 min. MRM was performed in positive ion mode using an AB SCIEX 6500+ QTRAP with the parameters below. GSH and LGSH were quantified using GSH-(glycine-13C2,15N). Samples were normalized to total protein.
| Q1 mass | Q3 mass | ID | DP (volts) | CE (volts) |
|---|---|---|---|---|
| 308.0 | 179.0 | GSH | 51.0 | 17.0 |
| 311.0 | 182.0 | GSH-(glycine-13C2,15N) | 51.0 | 17.0 |
| 380.1 | 233.1 | LGSH | 45.0 | 23.0 |
10
Resveratrol Quantification by LC-MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Resveratrol content was determined by LC/MS. Extracts analyzed by HPLC tandem mass spectrometry (LC-MS) were prepared in 0.1% formic acid. A Shimadzu LC system (Kyoto, Japan) was equipped with a Poroshell 120 EC-C18 (3.0 × 50 mm) column. The column temperature was fixed at 40 °C. Mobile phases were 0.1% formic acid in water (solvent A) and 0.1% formic acid in Acetonitrile (solvent B). The gradient was 0 min (60% A), 5 min (30% A), 8 min (5% A), 10 min (5% A), 10.01 min (60% A), and 15 min (60% A). This was followed by 0 min (40% B), 5 min (70% B), 8 min (95% B), 10 min (95% B), 10.01 min (40% B), and 15 min (40% B). The column effluent was monitored at 280 nm and mass spectra data were acquired by electrospray ionization (ESI) in the positive ion mode with a Tandem Mass Spectrometry(API 3200). The source temperature was 500 °C. LC-MS data were collected and processed by Analyst 1.6.2. (SCIEX) (Concord, Ontario, Canada).
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!