Antarctic krill PL was analyzed using a Shimadzu LC-30A system coupled to a Triple-TOF 6600 mass spectrometry with a 2.6 μm, 100 mm × 2.1 mm Kinetex C18 column (Phenomenex, Torrance, CA, USA). H2O/MeOH/ACN (1:1:1, v/v/v) and IPA/ACN (5:1, v/v) were used as mobile phase A and B, respectively, and both mobile phases contained 5 mM ammonium acetate. The binary gradient for PL separation was as follows: 0–0.5 min, 80% A, 0.5–1.5 min, 80–60% A, 1.5–3 min, 60–40% A, 3–8 min, 40–2% A, 8–10 min, and 2–80% A. The flow rate was set at 0.4 mL/min. The column temperature was set at 60 °C. After each analysis, the column was flushed with 80% of the mobile phase A for 5 min before the beginning of the next analysis. Data were collected in negative ionization mode. The collision energy of 30 eV was used for PA, 35 eV for other PLs.
Lc 30a system
Manufactured by Shimadzu
Sourced in Japan, United States
The Shimadzu LC-30A is a liquid chromatography system designed for high-performance liquid chromatography (HPLC) applications. It features a modular design that allows for the integration of various components, such as pumps, auto-samplers, and detectors, to create a customized analytical solution. The core function of the LC-30A system is to separate, identify, and quantify the components of a liquid sample.
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Lab products found in correlation
Lcms 8050 triple quadrupole lc ms ms, by Shimadzu (2 mentions)
Labsolutions software, by Shimadzu (2 mentions)
Lcms 8050, by Shimadzu (2 mentions)
Kinetex c18 column, by Phenomenex (1 mentions)
Acquity uplc beh c18 column, by Waters Corporation (1 mentions)
600 mhz spectrometer, by Bruker (1 mentions)
5600 q tof mass spectrometer, by AB Sciex (1 mentions)
Analyst tf software, by AB Sciex (1 mentions)
Tripletof 5600 mass spectrometer, by AB Sciex (1 mentions)
Tripletof 6600 mass spectrometer, by AB Sciex (1 mentions)
Inertsustain amide column, by GL Sciences (1 mentions)
Acquity upc2 system, by Waters Corporation (1 mentions)
13 protocols using lc 30a system
1
Lipidomic Analysis of Antarctic Krill
2
Quantification of TKIs by LC-MS/MS
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The amounts of TKIs were determined with a LCMS-8050 triple quadrupole LC-MS/MS (Shimadzu, Kyoto, Japan) coupled to an LC-30A system (Shimadzu) using an ACQUITY UPLC BEH C18 column (130 Å, 1.7 μm, ID 2.1 mm × 50 mm; Waters Corporation, MA, USA) at 40 °C. The mobile phase was composed of a mixture of 0.1% formic acid in water (pH 3.0) and 0.1% formic acid in acetonitrile at the flow rate of 0.2 mL/min. The mass numbers of the molecular and product ions for each compound were as follows: crizotinib (449.9 → 260.2, CE −24 V), gefitinib (447.2 → 128.2, CE −23 V), imatinib (494.3 → 394.3, CE −28 V) pazopanib (438.1 → 357.2, CE −35 V), sorafenib (464.9 → 252.0, CE −21 V), and sunitinib (399.2 → 283.0, CE −29 V). Labsolutions software (version 5.89, Shimadzu) was used for data manipulation. The detection limit was 10 nM for each compound.
3
Metabolite Analysis and Isolation in Aspergillus nidulans
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In order to perform small-scale metabolite analysis in A. nidulans, different transformants were grown on liquid CD-ST media at 28 °C for 3–5 days, then extracted with ethyl acetate (EtOAc). UPLC–MS analysis was carried out for 15 min on a Shimadzu LC-30A system connected to a single quadrupole mass spectrometer MS2020 (ESI) (Shimadzu, Kyoto, Japan), using a C18 reverse-phase column (shim pack XR-ODS III, 2.0 mm × 75 mm, 1.6 μm) with a linear gradient of 5−95% MeCN-H2O and a flow rate of 0.2 mL/min.
For the isolation of nanangelenin B (1 ), pseudofisnin A (2 ), and pseudofisnin B (3 ), the corresponding transformants of A. nidulans strains were grown on 4 L of liquid CD media for 4 days at 28 °C and then extracted with EtOAc. After concentration, the crude extract was chromatographed on a silica gel column using EtOAc and n-hexane as eluent. The fractions containing the target compounds were combined and further purified by semi-preparative HPLC, using a Pntulips C18 reverse-phase column (C18, 5 μm, 250 × 10 mm) (Puningtech, Shanghai, China).
The 1D and 2D NMR spectra were recorded in CDCl3 or DMSO-d6 using Bruker 600 MHz spectrometers and tetramethylsilane (TMS) as an internal standard. HR-ESIMS (high-resolution electrospray ionization mass) data were measured on an Acquity 2D-UPLC/Acquity UPC2/Xevo G2-XS QTOF (Waters, Milford, MA, USA).
For the isolation of nanangelenin B (
The 1D and 2D NMR spectra were recorded in CDCl3 or DMSO-d6 using Bruker 600 MHz spectrometers and tetramethylsilane (TMS) as an internal standard. HR-ESIMS (high-resolution electrospray ionization mass) data were measured on an Acquity 2D-UPLC/Acquity UPC2/Xevo G2-XS QTOF (Waters, Milford, MA, USA).
4
Flavonoid Metabolite Detection in db/db Mice
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In the current study, a TOF-MS/MS method was developed to detect flavonoid constituents and their metabolites in db/db mice. A Shimadzu LC-30A system (Kyoto, Japan) was used in this analysis. The mobile phase consisting of mobile phase A (0.1% formic acid-water) and mobile phase B (acetonitrile) was used for separation on an XSelect HSS T3 column (2.1 × 150 mm, 3.5 μm) with a flow rate of 0.3 mL/min. The following gradient elution program was used: 0–2 min, 5% B; 2–7 min, 9%–40% B; 7–12 min, 40%–65% B; 12–15 min, 65% B; 15–17 min, 65%–90% B; 17–20 min, 90% B; 20–22 min, 90%–5% B and 22–25 min, 5% B. The injection volume was 10 μL.
Mass spectrum data were acquired on AB Sciex 5600 Q-TOF mass spectrometer (Framingham, MA, United States) using negative information-dependent acquisition (IDA) modes. The source parameters were optimized and set as follows: MS and MS/MS scan mass range, m/z 100–1200 Da; gas 1, 60 Arb; gas 2, 65 Arb; curtain gas, 35 Arb; ion spray voltage, −4500 V; temperature, 550 °C; DP, −100 V; CE, −10 eV for MS scan, and 55 ± 25 eV for MS/MS scan. Real-time calibration was achieved by injecting APCI calibration solutions for every 5 samples. Analyst TF software (AB SCIEX) was used to acquire MS data.
Mass spectrum data were acquired on AB Sciex 5600 Q-TOF mass spectrometer (Framingham, MA, United States) using negative information-dependent acquisition (IDA) modes. The source parameters were optimized and set as follows: MS and MS/MS scan mass range, m/z 100–1200 Da; gas 1, 60 Arb; gas 2, 65 Arb; curtain gas, 35 Arb; ion spray voltage, −4500 V; temperature, 550 °C; DP, −100 V; CE, −10 eV for MS scan, and 55 ± 25 eV for MS/MS scan. Real-time calibration was achieved by injecting APCI calibration solutions for every 5 samples. Analyst TF software (AB SCIEX) was used to acquire MS data.
5
HPLC Analysis of Vitamins A and E
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High‐performance liquid chromatography analysis was performed on a SHIMADZU LC‐30A system equipped with diode‐array detector. Samples were analyzed using an ODS HYPERSIL C18 analytical column (3.0 µm, 100 mm × 2.1 mm) connected with a 2.1 mm × 20 mm guard column at a temperature of 30°C, and the flow rate was kept constantly at 0.5 mL/min. The mobile phase employed a gradient elution using the following constituents: Mobile phase A was water; mobile phase B was methanol. The gradient began with 93% methanol/water for 1 minutes, followed ramping to 100% methanol in 0.01 minutes. Then, 100% methanol was maintained for 2.5 minutes, then returned to initial conditions and equilibrated for 3.5 minutes. The autosampler temperature was 4°C, and the injection volume was 20 μL. The DAD detector was adjusted at 325 nm for vitamin A and vitamin A acetate, 291 nm for vitamin E and vitamin E acetate. The amount of individual vitamins was quantified from the corresponding peak area ratio of vitamin/internal standard (vitamin A/vitamin A acetate; vitamin E/vitamin E acetate) using LabSolutions software.
6
HPLC-MS Analysis of CHMACS Powder
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HPLC/MS was performed by Icast Testing Technology Service (Shanghai, China). Briefly, 0.2 g CHMACS freeze-dried powder was dissolved in 8 ml 50% methanol aqueous solution, extracted for 40 min by ultrasound at 45°C, and centrifuged at 13,000 rpm for 10 min. The supernatant thus obtained was filtered through a 0.22-μm microporous membrane. This sample (200 μl) was analyzed by HPLC/MS using, which was performed on a Shimadzu LC-30 A system with a C18 column (1.7 μm, 2.1 × 100 mm). The column temperature was 35°C, flow rate was 0.3 ml/min, and injection volume was 5 μl. Separation was achieved using the gradient elution procedure and mobile phase, which consisted of acetonitrile–0.2% formic acid aqueous solution. MS was performed on an AB Sciex Triple TOF™ 5600 mass spectrometer, which was equipped with an electrospray ionization source in the positive ion mode. The voltage of the ion source was 5500 V, temperature was 500°C, de-cluster voltage was 100 V, collision energy was 35 eV, and collision energy spread was 15 eV. The first-order mass spectrum parent ion scanning Fan Tian was 50:1000, and the IDA setting response Bo exceeded the six peaks of 100 cps for secondary MS scanning. The sub-ion scanning range was 50–1000, and the open-state back deduction.
7
Untargeted Metabolomics of Liver Samples
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Extraction of metabolites was performed from the liver samples (∼20 mg). The TripleTOF 6600+ mass spectrometer (SCIEX, Foster City, CA, USA), coupled with an LC-30A system (Shimadzu, Kyoto, Japan), was used for the untargeted metabolomics profiling by liquid chromatography–tandem mass spectrometry (LC-MS/MS) system. On a Waters ACQUITY Premier HSS T3 Column 1.8 µm (2.1 mm * 100 mm ) at 40 °C, chromatographic separation was accomplished. At a flow rate of 0.4 mL/min, the mobile phase was composed of 0.1% formic acid/water (A) and 0.1% formic acid/acetonitrile (B). The database of the Metware Biotechnology Co., Ltd. (Wuhan, China) and multiple reaction monitoring (MRM) were the bases for the qualitative and quantitative study of metabolites.
8
Quantification of 1-Methylnicotinamide by LC-MS/MS
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The concentration of 1-MNA was determined using an LC-MS-8050 triple quadrupole LC-MS/MS (Shimadzu, Kyoto, Japan) coupled to an LC-30A system (Shimadzu). Chromatography was performed on an InertSustain amide column (ID 2.0 mm × 50 mm; GL Sciences) at 40 °C by means of step-gradient elution (flow rate, 0.4 mL/min) as follows: 0 to 2.0 min, 13% A/87% B; 2.0 to 2.5 min, 13% A/87% B to 50% A/50% B; 2.5 to 4.0 min, 50% A/50% B; 4.0 to 4.5 min, 50% A/50% B to 13% A/87% B; and 4.5 to 7.0 min, 13% A/87% B. A was water containing 0.1% formic acid and B was acetonitrile containing 0.1% formic acid. The mass numbers of the molecular and product ions for each compound were as follows: 1-MNA (137.2 → 94.2, CE − 22.0 V) and 1-MNA-d3 (Toronto Research Chemicals, North York, Canada) (140.0 → 97.2, CE −22.0 V). Lab solutions software (version 5.89, Shimadzu) was used for data manipulation. The detection limit was 10 ng/mL for each compound.
9
Quantitative Analysis of Compounds by LC-MS/MS
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Individual compounds concentrations were measured by LC-MS/MS (LCMS-8050; Shimadzu, Kyoto, Japan) coupled to an LC-30A system (Shimadzu). The analytical column was a Luna 5 µm C18(2) LC Column (50 × 2 mm; Phenomenex, CA, U.S.A.) maintained at 40 °C. The mobile phase was composed of a 0.1% formic acid aqueous solution (solvent A) and acetonitrile containing 0.1% formic acid (solvent B). The flow rate was 0.4 mL/min and the injection volume was 5 µL. The gradient profile was as follows: 5% solvent B for 1.0 min; linear ramp to 95% solvent B in 3.0 min; then return to initial conditions in 0.5 min. The detection conditions of each compound by LC-MS/MS are summarized in Table 1. The lower limit of detection for each compound was 1 nM.
Data Analyses The statistical significance of differences in mean values was determined using a Student's t-test. Differences with a p-value less than 0.05 were considered statistically significant.
Data Analyses The statistical significance of differences in mean values was determined using a Student's t-test. Differences with a p-value less than 0.05 were considered statistically significant.
10
Quantitative Analysis of Asperulosidic Acid
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The quantitative analysis was performed by an LC-30A system (Shimadzu, Japan). The wavelength used of the detector was 238 nm. The proportion of the mobile phase was the same as UHPLC-MS. The UHPLC-UV method was validated for accuracy, precision, specificity, linearity, and range according to the guidelines of the Chinese Pharmacopeia (CH.P), 2015, chapter 9101.
Accuracy of the method is calculated by a recovery test. The accurate amount of asperulosidic acid standard solutions with three different concentration levels was added to sample and three replicates of each concentration. The average recoveries were determined by the formula:
The precision tests were performed using six replicated injections of the same sample solution in a day.
The calibration curves of asperulosidic acid for quantitation were created by establishing a relationship between the peak area (Y) and the concentration (X, µg/ml) of the standard solution. Stored standard solution of asperulosidic acid (364 μg ml−1) was diluted to 7.28 μg ml−1, 14.56 μg ml−1, 29.12 μg ml−1, 58.24 μg ml−1, and 116.48 μg ml−1 for the UHPLC-UV test.
Accuracy of the method is calculated by a recovery test. The accurate amount of asperulosidic acid standard solutions with three different concentration levels was added to sample and three replicates of each concentration. The average recoveries were determined by the formula:
The precision tests were performed using six replicated injections of the same sample solution in a day.
The calibration curves of asperulosidic acid for quantitation were created by establishing a relationship between the peak area (Y) and the concentration (X, µg/ml) of the standard solution. Stored standard solution of asperulosidic acid (364 μg ml−1) was diluted to 7.28 μg ml−1, 14.56 μg ml−1, 29.12 μg ml−1, 58.24 μg ml−1, and 116.48 μg ml−1 for the UHPLC-UV test.
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