Bioactive fractions were subjected to a two-step RP-HPLC procedure conducted on an Agilent 1100 Series HPLC Value system (Agilent Technologies, Hewlett-Packard-Strasse 8, Germany) using a Peptide XB-C18 10 (5 µm, 250 × 4.6 mm, Aeris™, Canada). For these two steps of purification, the mobile phase consisted of buffer A (H2O, 5mM HCl) and buffer B (ACN). The column was heated at 40 °C and elution was performed using a linear gradient of buffer B at 0.8 mL/min. The gradient program started with 70% of buffer B for 3 min before increasing to 90% over 20 min. Solvent was maintained at 90% of buffer B for 1 min before returning to starting conditions over 2 min. The injection volume was 100 µL and the elution was monitored by UV absorbance at 214 nm. Every 5 min, fractions were collected and then dried. Before being tested for their antibacterial activity, the fractions were dissolved in water with acetonitrile (50:50, v/v).
Agilent 1100 series hplc value system
Manufactured by Agilent Technologies
Sourced in United States, Germany
The Agilent 1100 Series HPLC Value System is a high-performance liquid chromatography (HPLC) instrument designed for laboratory use. It provides a robust and reliable solution for analytical separation and quantification of compounds in complex samples. The system includes essential components such as a pump, autosampler, column compartment, and detector, enabling efficient and reproducible liquid chromatography analysis.
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Lab products found in correlation
Eclipse xdb c18 column, by Agilent Technologies (1 mentions)
Eclipse aaa column, by Agilent Technologies (1 mentions)
Voyager de pro, by Thermo Fisher Scientific (1 mentions)
Openlab cds chemstation edition, by Agilent Technologies (1 mentions)
Advia 2120, by Siemens (1 mentions)
Capcellpak ug120 c18 column, by Shiseido (1 mentions)
8 protocols using agilent 1100 series hplc value system
1
Two-Step RP-HPLC Purification of Bioactive Fractions
2
Quantification of Surfactin by HPLC
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The supernatants from the centrifuged cell culture samples were filtered (0.2 µm) and the surfactin concentration was determined by reversed-phase HPLC (Agilent 1100 Series HPLC Value System, Agilent Technologies, Diegem, Belgium) with an Eclipse XDB C−18 column (3.5 µm, 2.1 × 150 mm) (Agilent Technologies, Diegem, Belgium). The HPLC analysis method was based on an isocratic elution profile with a mobile phase composition of 80% acetonitrile and 20% water containing 0.1% trifluoroacetic acid (TFA). The flow rate was set at 0.4 mL min−1 with an analysis time of 22 min per sample. The surfactin molecules were detected by UV at 214 nm. Purified surfactin samples (>98%) (Lipofabrik, Villeneuve d’Ascq, France) were injected to identify the retention time of the surfactin molecules and to determine a calibration curve.
3
MALDI-TOF MS and HPLC Analysis of PLHs
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The molecular weight distribution of PLHs was determined according to the methods of Aspmo et al [24 ] with some modifications. Samples (1 μL) were analyzed using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS; Voyager DE-PRO, Applied Biosystems, NY, USA) in a positive ion mode, whereas the results were obtained in the linear mode. The amino acid composition of the freeze-dried PLHs was determined using a modified method of You and Wu [25 (link)]. Briefly, 0.1 g of PLHs was hydrolyzed with 3 mL of 4 M methanesulfonic acid, containing 0.2% 3-(2-aminoethyl) indole under vacuum at 115°C for 48 h. The total amino acids were analyzed using an Agilent 1100 series high performance liquid chromatography system (Agilent 1100 Series HPLC Value System, Agilent Technologies, Santa Clara, CA, USA) equipped with ZORBAX Eclipse-AAA column (4.6×150 mm, 3.5 μm).
4
Quantification of Phenolic Compound PCA
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In order to quantify PCA, the samples were analysed using high performance liquid chromatography (HPLC) (Agilent 1100 Series HPLC Value System, Agilent Technologies). Analyses were performed using a Synergi™ 4 μm Hydro-RP 80 Å, 100 × 2 mm column (Phenomenex, Torrance, CA, USA). For the analysis, 50 μL of PCA solution were injected and tested at a mobile phase flow rate of 0.4 mL/min. The following gradient system was used: solvent A—0.1% FA in water, solvent B—0.1% FA in acetonitrile; 0 min—95% A and 5% B, 15 min—0% A and 100% B, 15.01 min—95% A and 5% B.
PCA concentrations were calculated using the software OpenLAB CDS ChemStation Edition version A0.2.13 (Agilent Technologies). The PCA amounts computed by the software were corrected for the dilution factor and the potato samples’ mass.
PCA concentrations were calculated using the software OpenLAB CDS ChemStation Edition version A0.2.13 (Agilent Technologies). The PCA amounts computed by the software were corrected for the dilution factor and the potato samples’ mass.
5
Bcl-2 Levels in β-Thalassemia Minor
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Ninety-seven patients (60 females and 37 males with mean age of 29±21 years) with β-thalassemia minor were enrolled in this study. The diagnosis of β-thalassemia minor was based on whole blood counts, family history, and HbA2 levels estimated by high-performance liquid chromatography (Agilent 1100 Series HPLC Value System, Waldbronn, Germany). The control group comprised 23 healthy adults (17 females and 6 males with mean age of 58±9 years) without anemia. The levels of serum Bcl-2 were measured using a commercial enzyme-linked immunosorbent assay kit (Biosource, Cat. No. TMA 0311, Camarillo, CA, USA). Age, sex, hemoglobin, hematocrit, mean corpuscular volume (MCV), whole blood cell counts, and serum levels of Bcl-2 were recorded. Venous blood samples were taken under the supervision of medical personnel and were measured with an ADVIA 2120 instrument (Siemens, Erlangen, Germany). Signed informed consent was obtained from all participants. SPSS 15 (SPSS Inc., Chicago, IL, USA) and the Mann-Whitney U test were used in statistical evaluation of data and p<0.05 was accepted as statistically significant.
6
Rotigotine Solubility Screening for Microemulsions
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To identify the optimal components for the microemulsion formulations, the solubility of rotigotine in different oils (olive oil, Labrafil M 1944 CS, Labrafac Lipophile and oleic acid, and Tween 80), surfactants (Labrasol, Cremophor RH40, and Transcutol HP), and cosurfactants (PEG-400, 1,2-propylene glycol, and absolute ethyl alcohol) was measured. An excess amount of rotigotine was added to a fixed volume of each solvent (10 mL), and the mixture was incubated at 25°C in a constant temperature water bath oscillator (BoXun SHZ-A, Shanghai, People’s Republic of China) for 24 hours. The resulting suspension was then centrifuged for 10 minutes at 12,000 rpm. After appropriate dilution, the supernatant was filtered using a 0.45 μm microporous membrane filter. The concentration of rotigotine in the filtrate was determined by high-performance liquid chromatography (HPLC) (Agilent 1100 Series HPLC Value System; Agilent Technologies, Santa Clara, CA USA) with detection at a wavelength of 223 nm. The solubility of rotigotine in the various solvents was then calculated to identify high-solubility solvents.
7
Quantitative Analysis of Nucleotides in Milk
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Laboratory instruments were used for general quantitative analysis. Whatman No.1
filter paper (Maidstone, UK) was used for filtration and Macherey-Nagel
Chromabond SB 6 mL/1,000 mg column (Duren, Germany) was used for SPE. The vacuum
manifold used was from Supelco (Maidstone, UK). To filter the final extracted
solution, a 0.22-μm nylon filter (Adventec, Japan) was used. The Agilent
1100 Series HPLC Value System (Agilent, Santa Clara, CA, USA) and the Agilent
1200 Series DAD HPLC System (Santa Clara, CA, USA) were used. The analytical
conditions of the instrument (HPLC-DAD) were optimized prior to initial use. To
effectively detect nucleotides in milk powder and human milk, the sensitivity
was analyzed using DAD, and the Capcellpak UG 120 C18 column
(4.6×250 mm, 5 cm, Shiseido, Japan) was used for separation of
nucleotides. Five optimum separation conditions were used. The mobile phase used
was KH2PO4 (10 mM, pH 5.6), flow rate was 0.6 mL/min,
column temperature was 40°C, total analysis time was 25 min, and
injection volume was 10 μL.
filter paper (Maidstone, UK) was used for filtration and Macherey-Nagel
Chromabond SB 6 mL/1,000 mg column (Duren, Germany) was used for SPE. The vacuum
manifold used was from Supelco (Maidstone, UK). To filter the final extracted
solution, a 0.22-μm nylon filter (Adventec, Japan) was used. The Agilent
1100 Series HPLC Value System (Agilent, Santa Clara, CA, USA) and the Agilent
1200 Series DAD HPLC System (Santa Clara, CA, USA) were used. The analytical
conditions of the instrument (HPLC-DAD) were optimized prior to initial use. To
effectively detect nucleotides in milk powder and human milk, the sensitivity
was analyzed using DAD, and the Capcellpak UG 120 C18 column
(4.6×250 mm, 5 cm, Shiseido, Japan) was used for separation of
nucleotides. Five optimum separation conditions were used. The mobile phase used
was KH2PO4 (10 mM, pH 5.6), flow rate was 0.6 mL/min,
column temperature was 40°C, total analysis time was 25 min, and
injection volume was 10 μL.
8
N-Acetylaspartate Measurement in MS
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