Belinostat was analyzed by high-performance liquid chromatography (HPLC). The HPLC system consisted of an L-7100 pump, L-7200 autosampler, L-7455 diode array detector at 265 nm (Hitachi, Tokyo, Japan) and a Purospher Star RP-18 endcapped column (250 × 4.6 mm, internal diameters 5 μm, Merck). The mobile phase was a mixture of 0.1% phosphoric acid and acetonitrile by gradient elution, and the flow rate was 1 mL/min. The column oven was set at 30 °C. Limits of detection and quantitation of belinostat were determined by dissolving belinostat at decreasing concentrations in distilled deionized water until the signal/noise ratios were 3 and 10, respectively. The linearity of the standard curves and intraday and interday precision and accuracy were established.
L 7200 autosampler
Manufactured by Hitachi
Sourced in Japan
The L-7200 autosampler is a laboratory equipment device designed for automated sample handling and injection. Its core function is to precisely and consistently introduce liquid samples into an analytical instrument, such as a high-performance liquid chromatograph (HPLC) or gas chromatograph (GC), for analysis.
Automatically generated - may contain errors
Lab products found in correlation
L 7100 pump, by Hitachi (7 mentions)
D 7000 interface, by Hitachi (3 mentions)
L 7455 detector, by Hitachi (2 mentions)
Hplc system, by Hitachi (2 mentions)
L 7455 diode array detector, by Hitachi (1 mentions)
Purospher star rp 18 endcapped column, by Merck Group (1 mentions)
Hctultra ptm discovery system, by Bruker (1 mentions)
L 7400 uv detector, by Hitachi (1 mentions)
Securityguard c18 guard column, by Phenomenex (1 mentions)
Eclipse xdb c8, by Agilent Technologies (1 mentions)
Mightysil rp 18 column, by Kanto Chemical (1 mentions)
L 7485 fluorescence detector, by Hitachi (1 mentions)
L 7300 column oven, by Hitachi (1 mentions)
7 protocols using l 7200 autosampler
1
HPLC Analysis of Belinostat
2
HPLC Analysis of DHM in APF Complex
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Analytical HPLC was performed on Hitachi D-7000 interface equipped with L-7100 pump, L-7455 detector and L-7200 autosampler (Tokyo, Japan) to determine DHM content in APF complex. The test solution was prepared by mixing 10 mg of APF complex with 50 mL of methanol under ultrasonic condition at room temperature for 40 min followed by filtration through a 0.45 μm filter. The standard solution was prepared by mixing DHM (the purity is higher than 98%, Sigma Chemical Co., St. Louis, MO) with methanol to obtain different concentrations through serial dilution. Chromatographic separation was carried out on a Mightysil RP-18 column (250 × 4.6 mm, 5 μm) using a gradient solvent system comprised of acetonitrile (A) and 0.03% (v/v) H3PO4 (B). Gradient profile was set as follows at 0–10 min with the ratio of 15% A and 85% B; at 10–20 min with the ratio of 15–25% A and 85-75% B; at 20–25 min with the ratio of 25–40% A and 75-60% B; at 25–40 min with the ratio of 40–15% A and 60–85% B. The UV wavelength, flow rate, and injection volume were set at 210 nm, 1.0 mL/min, and 10 μL, respectively.
3
HPLC Analysis of Target Compounds
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Our previously described method was slightly modified (Pan et al., 2010 ). Before HPLC analysis, MEMO was filtered with a 0.2 µm Millipore filter. A total volume of 10 μL was loaded to the column. External standards of 100 μg/mL were prepared with HPLC grade‐methanol for calculation of the concentrations of examined samples. Reverse phase HPLC was performed on a HITACHI HPLC system (Tokyo, Japan) equipped with HITACHI L‐7100 pump, HITACHI L‐7400 UV detector, and HITACHI L‐7200 autosampler. LiChroCART 250–4 C18 HPLC cartridge (5 μm; Merck, Whitehouse Station, NJ, USA) was used for separations. Conditions for separation of examined samples were listed in Table 1 . Based on the separation conditions of the standard, two candidate fractions were harvested (collection period: retention time ± 5 min). These fractions were further determined by electrospray ionization‐ion trap mass spectrometry (HCT ultra PTM Discovery system; Bruker Daltonics, Billerica, MA, USA) for identification of the target compounds. Capillary voltage was 4,000 V, capillary exit offset 220 V, skimmer potential 60 V, and the trap drive value was 78. Conventional data of electrospray ionization‐ion trap mass spectrometry were recorded using a scan range of 150–280 m/z. Nebulizer (nitrogen) pressure was 10 psi with dry gas flow of 5 L/min and dry temperature at 300°C.
4
HPLC Analysis of Betalain Pigments
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Betalain composition analyses were carried out as described by Cai et al. (2001a) (link). An HPLC-system (Hitachi High Technologies America, Inc., Schaumburg, IL), equipped with a L-7200 autosampler, D-7000 interface module, L-7100 pump, and a L-7455 diode array detector was used for pigment analyses. Separation was achieved using a LUNA Phenomenex C18 reversed-phase column (5 μm, 250 × 4.6 mm, Torrance, CA) and a SecurityGuard C18 guard column (Phenomenex; Torrance, CA). The mobile phase used 1.5% (v/v) phosphoric acid as solvent A and 1.5% phosphoric acid, 20% acetic acid, and 25% acetonitrile for solvent B. A 30-min linear gradient from 10 to 55% solvent B in solvent A was used, with an added 1-min 100% B wash followed by a 5-min equilibration at 10% B after each sample. An injection volume of 20 μl was used with monitoring at 535 nm. The column was kept at ambient temperature at a constant flow rate of 1 ml/min and samples were injected via a 20 μl sample loop. Purified betanin (Millipore-Sigma; St. Louis, MO) was used as a standard for compound identification with each set of samples analyzed, and identified compounds were quantified as a percentage of the total betacyanin content determined previously using a UV–Vis spectrophotometer. Spectral DAD data were simultaneously collected between 250 and 600 nm.
5
HPLC Analysis of Organic Compounds
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Hitachi LaChrom series LC system, consisting of an L-7100 pump, an L-7200 autosampler, an L-7400 UV detector set at a wavelength of 280 nm, and D-7000 interface with system manager data acquisition software (version 5.0), was used throughout the study. The chromatographic separation was achieved using an Agilent Eclipse XDB C8 (150 × 4.6 mm, 5 μ) column, the optimized method used a segmented gradient mobile phase with phosphate buffer at pH = 2.5 as solvent (A) and ACN as solvent (B), and the gradient program is shown in Table 1 . The sample volume for injection was 50 μL and the total run time was 20 min.
6
Quantitative Analysis of Herbal Compounds
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Analytical high-performance liquid chromatography was performed on Hitachi D-7000 Interface equipped with L-7100 pump, L-7455 detector, and L-7200 autosampler (Hitachi, Tokyo, Japan) to determine the marker compounds in three herbal mixture extracts. For the analysis of dihydromyricetin, salvianolic acid B, and EGCG content in T1, T2, and T3 complexes, respectively, the test solution was prepared by mixing 10 mg of dried powder with 10 mL of 70% (v/v) methanol under ultrasonic condition at room temperature for 1 hour followed by filtration. These filtrates were filtrated through a 0.45-μm filter and chromatographic separation was carried out on a Mightysil RP-18 column (Kanto Chemical CO., INC., Tokyo, Japan.) with the specification of 250mm × 4.6 mm, 5 μm using gradient solvent systems as summarized in Table 2 . The UV wavelength, flow rate and injection volume were set at 254 nm, 1.0 mL/min, and 10 μL, respectively. The standard solution was prepared by mixing dihydromyricetin, salvianolic acid B, or EGCG, respectively, with methanol to obtain different concentrations through serial dilution.
7
Analytical HPLC for Fluorescent Compound
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Analytical HPLC was performed on Hitachi D-7000 interface equipped with L-7485 fluorescence detector, L-7100 pump, L-7200 autosampler, L-7300 column oven, and post-column photochemical reaction system (Tokyo, Japan). Chromatographic separation was carried out on a Mightysil RP-18 column (250 × 4.6 mm, 5 μm). HPLC conditions were established according to the research from Trucksess et al. [5 (link)] with minor modification. Briefly, the wavelengths for excitation and emission were 360 nm and 440 nm, respectively. The mobile phase, flow rate, injection volume, column temperature, and stop time were set at 45% (v:v) methanol, 1.0 mL/min, 20 μL, 40°C, and 20 min, respectively.
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