Samples of oleuropein derived from entrapment efficacy and release profile were analyzed using HPLC (A Jasco PU-1580 intelligent HPLC pump, Tokyo, Japan). The chromatographic system was equipped with a UV photodiode detector (multiwavelength) (Jasco MD 1510 diode, Tokyo, Japan). The column C18 in reversed-phase (250 by 4.60 mm–5.0 µm), was maintained at room temperature. Other chromatographic conditions were as follows: Mobile phase was a 25:75 (v/v) mixture of acetonitrile:water and flow rate was 1 mL/min [20 (link)]. The UV detection wavelength was 230 nm.
Pu 1580 intelligent hplc pump
Manufactured by Jasco
Sourced in Japan
The PU-1580 intelligent HPLC pump is a high-performance liquid chromatography pump designed for analytical and preparative applications. It features precise flow control, advanced programming capabilities, and reliable operation. The pump can deliver flow rates ranging from 0.001 to 10 mL/min with a flow precision of ±0.1%.
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Lab products found in correlation
Accq tag eluent a concentrate, by Waters Corporation (1 mentions)
Accq fluor reagent kit, by Waters Corporation (1 mentions)
Intelligent fluorescence detector fp 1520, by Jasco (1 mentions)
As 2055 plus intelligent sampler, by Jasco (1 mentions)
Accq fluor reagent, by Waters Corporation (1 mentions)
Accq fluor borate buffer, by Waters Corporation (1 mentions)
Avanti 30 centrifuge, by Beckman Coulter (1 mentions)
4 protocols using pu 1580 intelligent hplc pump
1
HPLC Analysis of Oleuropein Release
2
Amino Acid Analysis of DM Media
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For the variation of amino acid concentrations in DM media, samples were subjected to an AccQ-Fluor Reagent (AQC, 6-aminoquinolyl-N-hydroxysuccinimide carbamate) derivatization (Waters Corp., Milford, MA, United States) according to the manufacturer’s protocol. AQC was reconstituted at a final concentration of 10 mM in acetonitrile, included in the AccQ-Fluor Reagent Kit (Waters Corp.). Briefly, 10 μL of samples were derivatizated with 70 μL of AccQ-Fluor Borate Buffer (Waters Corp.) and 20 μL of reconstituted reagent. The samples were heated to 55°C for 10 min. The amino acids content was analyzed using an HPLC (PU-1580 Intelligent HPLC pump, Intelligent Fluorescence Detector FP-1520 and Intelligent Sampler AS-2055 Plus, with 10 μl loop; Jasco Corp.). Separation of amino acids was obtained using AccQ-TagTM column (3.9 mm × 150 mm) for amino acid analysis (Waters Corp.). A gradient elution was performed maintaining a column temperature of 30°C and using two mobile phases: A (100 ml of AccQ-Tag Eluent A concentrate (Waters Corp.), diluted 1:10 with H2O for chromatography (Sigma-Aldrich, St. Louis, MO, United States) and B (60% acetonitrile and 40% H2O for chromatography) (Sigma-Aldrich, St. Louis, MO, United States) with a flow rate of 1 ml/min. The fluorescence detector was set at excitation wavelength of 250 nm and emission wavelength of 395 nm.
3
Quantifying Sulforaphane Encapsulation in Deformable Vesicles
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The amount of entrapped sulforaphane was separated from the untrapped aliquot by using ultracentrifugation. The colloidal vesicles were poured into polycarbonate tubes and then centrifuged at 90,000× g for 1 h at 4 °C using an Avanti 30 Centrifuge (Beckman, Fullerton, CA, USA) equipped with a fixed angle rotor Beckman mod. F1202. The supernatant and the pellet were divided and separately analyzed using HPLC (A Jasco PU-1580 intelligent HPLC pump, Tokyo, Japan) (see Section 2.9 ).
To break the pellets, 4 mL of ethanol was used. Possible interference from vesicle components was avoided using empty ethosomes® and transfersomes® as blanks. To quantify the amount of sulforaphane entrapped in vesicular systems, the difference between the drug used during the preparation and the non-encapsulated drug was calculated. The following equation was used to determine the encapsulation efficiency (EE%) of sulforaphane in deformable vesicles:
where De is the amount (mg) of entrapped sulforaphane and Da is the sulforaphane amount (mg) used to prepare ethosomal and transfersomal formulations. The reported results represent the average value of five different formulations ± standard deviation.
To break the pellets, 4 mL of ethanol was used. Possible interference from vesicle components was avoided using empty ethosomes® and transfersomes® as blanks. To quantify the amount of sulforaphane entrapped in vesicular systems, the difference between the drug used during the preparation and the non-encapsulated drug was calculated. The following equation was used to determine the encapsulation efficiency (EE%) of sulforaphane in deformable vesicles:
where De is the amount (mg) of entrapped sulforaphane and Da is the sulforaphane amount (mg) used to prepare ethosomal and transfersomal formulations. The reported results represent the average value of five different formulations ± standard deviation.
4
Cocoa Butter Extraction and Salad Dressing Formulation
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Dried cocoa beans were purchased from Lembaga Koko Malaysia (Jengka, Malaysia). Cocoa shells were removed manually from the beans and nibs were smashed with a mortar and pestle to approx. <1 mm in size. Cocoa butter used in this study was extracted from the nibs using supercritical CO2 extraction and ethanol. For supercritical extraction we used model PU-1580 Intelligent HPLC pump (Jasco Corporation, Tokyo, Japan). A back pressure regulator model BP-1580-81 (Jasco Corporation) controlled the extraction pressure and separation. Approximately 20 g of cocoa nibs were loaded into 50-mL extraction vessel. Extraction with ethanol occurred at 78 °C, 35 MPa pressure and a flow rate of 2 mL/min. The brand of commercial salad dressing was Lady's Choice MayoLite (Unilever, Kuala Lumpur, Malaysia). The ingredients include soybean oil, egg, modifed starch, sugar, salt, vinegar, lemon juice, potassium sorbate, phosphoric acid, citric acid, calcium disodium EDTA and edible gum. Newly formulated dressing was composed of soybean oil (Soya Lite, Subang Jaya, Malaysia), lemon juice, egg yolk, sodium benzoate, vinegar, sugar, salt, and xanthan and Arabic gum (both from Markaids Sdn. Bhd., Selangor, Malaysia).
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