Loading capacity was determined by using a Dialysis tubing cellulose membrane (sigma-Aldrich, Massachusetts, USA) against PBS by adding 2 ml of the mixture after probe-sonication into a cellulose dialysis sac bag that was cut off by scissors then the dialysis bag was sealed properly both from top and bottom and inserted into PBS 7.4 with 50 rpm using a shaking incubator at the room temperature. After 0.5h, the media was collected and replaced with new PBS for another 0.5h. Then the media collected and standard dilutions were filtered by syringe and tested using a Waters 2690 Alliance HPLC system equipped with a Waters 996 photodiode array detector. Standard preparation: Weight 1 mg of each standard to obtain stock solution then serial dilution to obtain conc. of (100,80,60,40,20,10 µg/ml) then filtrated using a 0.22 syringe filter and injected 10 µl. Sample preparation: 1) The sample is then filtrated using a 0.22 syringe filter and injected 10 µl. HPLC analysis conditions were: Column Kromasil C8: 4.6 × 250 mm, 5 µm, Mobile phase: 0.1% Formic acid in water: Acetonitrile, Mode of elution: isocratic, Flow rate: 1ml/min, Temperature: Ambient, Wavelength: 254 nm22 (link).
2690 alliance hplc system
Manufactured by Waters Corporation
Sourced in United States
The 2690 Alliance HPLC system is a liquid chromatography instrument designed for high-performance liquid chromatography (HPLC) analysis. It is capable of separating complex mixtures of compounds and providing precise quantitative and qualitative data. The system features automated sample handling, precise solvent delivery, and advanced data processing capabilities to support a wide range of analytical applications.
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Lab products found in correlation
996 photodiode array detector, by Waters Corporation (6 mentions)
Waters 996 photodiode array detector, by Waters Corporation (2 mentions)
Laminar flow, by Thermo Fisher Scientific (2 mentions)
Nova pak c18, by Waters Corporation (2 mentions)
Dialysis tubing cellulose membrane, by Merck Group (1 mentions)
Jtem model 1010, by JEOL (1 mentions)
Malvern zetasizer nano z, by Malvern Panalytical (1 mentions)
Uv 1601, by Shimadzu (1 mentions)
V 630, by Jasco (1 mentions)
Rotavapor rii, by Büchi (1 mentions)
Uv vis 2487 detector, by Waters Corporation (1 mentions)
Cosmosil 5c18 ar 2, by Nacalai Tesque (1 mentions)
Cosmosil π nap, by Nacalai Tesque (1 mentions)
Symmetry shield c18, by Waters Corporation (1 mentions)
Nucleosil ods reversed phase column, by Waters Corporation (1 mentions)
Empower 2, by Waters Corporation (1 mentions)
2996 pda detector, by Waters Corporation (1 mentions)
19 protocols using 2690 alliance hplc system
1
Dialysis-based Loading Capacity Determination
2
HPLC Analysis of Phytochemical Constituents
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A Waters 2690 Alliance HPLC system was equipped with a Waters 996 photodiode array detector. The extract (rose MeOH extract, 93 mg/mL) was dissolved in methanol and filtered using a 0.22 μm syringe filter, and 10 μL was injected. Stock solutions of 10 different standards in methanol were prepared. Each of the standards was filtered using a 0.22 μm syringe filter, and then, 10 μL was injected. HPLC analysis conditions were as follows: Column C18, Inertsil ODS 3: 4.6 × 250 mm, 5 μm; mobile phase (A): buffer (0.1% phosphoric acid in water) and methanol (65% water: 35% methanol); mobile phase (B): buffer (0.1% phosphoric acid in water) and methanol (50% water: 50% methanol); mode of elution: gradient; flow rate: 1 mL/min; temperature: ambient; and wavelength: 280 nm. The HPLC conditions and optimisation timetable are shown in Table 1 .
3
Characterization of Silver Nanoparticles
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A rotary evaporator (Rotavapor R-II, Buchi, Switzerland) was utilized for the solvent evaporation process. The antioxidant assays, as well as quantification assays, were conducted using a spectrophotometer (UV-1601, Shimadzu, Japan). The qualitative identification of phenolic compounds was performed using Waters 2690 Alliance HPLC system and Waters 996 photodiode array detector, Milford, CT, USA. Laminar Flow (model 1386, Thermo Fisher Scientific, Waltham, MA, USA) and BIO-RAD microplate reader (model iMark, Japan) were utilized. Centrifugation of the AgNPs was done using a cooling centrifuge (PRO-Research K241R; Centurion, West Sussex, UK), characterization of the prepared NPs was done using a double-beam spectrophotometer (V630, Jasco, Tokyo, Japan), transmission electron microscope (TEM) (JTEM model 1010, JEOL®, Tokyo, Japan) and Malvern Zetasizer (Nano ZS, Malvern Instruments Ltd., Malvern, UK).
4
Isolation and Quantification of Egg Yolk Carotenoids
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Six eggs per group were placed in −20°C for 24 h where egg white was coagulated and was easily separated from egg yolk. The latter was smashed by a spatula and 1 g of egg yolk was transferred to an amber-colored vial and stirred with 20 mL acetone for 15 min. The mixture was then incubated at −20°C for 48 h to precipitate proteins. After the incubation period, the supernatant was collected and dried under vacuum at 30°C. The dried residue was re-dissolved in 2 mL acetone of high-performance liquid chromatography (HPLC ) grade for HPLC analysis (Schierle et al., 2003 ). The sample was filtered using a 0.22-µm syringe filter. HPLC (Waters 2690 Alliance HPLC system equipped with a Waters 996 photodiode array detector) quantified the concentration of lycopene and lutein. Lutein stock solution of 100 µg/mL in acetone of HPLC grade was prepared, and 6 serial dilutions were prepared in the concentrations of 70, 60, 50, 40, 30, and 20 µg/mL. Then, the following concentrations were prepared from the previous stock solution: 25, 17.5, 12.5, and 10 µg/mL in acetone of HPLC grade. Stock solution of 100 µg/mL was prepared from which the following concentrations were established: 35, 31.5, 28, 24.5, and 17.5 µg/mL in acetone of HPLC grade (Nimalaratne et al., 2013 ).
5
HPLC Analysis of Methanolic Samples
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This was performed according to the method of Singh et al. [25 (link)]. Each of the samples and 10 different standards solutions were dissolved in methanol and filtered using 0.22 µm syringe filters; then, 100 µL of each sample and 10 µL of each standard were injected using HPLC column, Waters 2690 Alliance HPLC system equipped with a Waters 996 photodiode array detector. Column C18 Inertsil ODS 3: 4.6 × 250 mm, 5 µm, and mobile phase: Buffer (0.1% phosphoric acid in water) and methanol mode of elution was gradient with a flow rate of 1 mL/min at wavelength 280 nm.
6
Cellular Uptake of Leflunomide in MDA-MB-231 Cells
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MDA-MB-231 cell line was used to study the cellular uptake of the provided LEF and the selected LEF loaded cubosomal formulation. On the first day, the cells were seeded in T25 flask. On the second day, the media were removed, and media supplemented with predetermined drug concentration were added to cells. A confluent T25 flask 0.7 × 106 of MDA-MB-231 cells was harvested after 6 and 24 hours of incubation. The pellets were collected at each time interval were subjected to lysis according to the method previously reported by Srisongkram et al. (Srisongkram & Weerapreeyakul, 2019 (link)). Sample preparation for HPLC analysis was carried out as follows: 0.5 mL methanol was added to each sample then sonicated for 15 min. This step was followed by centrifugation for 10 min at 4500 rpm and filtration using 0.22 μm Nylon syringe filter.
Amount of LEF taken by the cells were quantified using HPLC (Waters 2690 Alliance HPLC system equipped with a Waters 996 photodiode array detector). The Mobile phase consisted of (50%:50%) acetonitrile: phosphate buffer (0.02 M) pH was adjusted to 2.6 with orthophosphoric acid. Flow rate was 1 mL/min and the measurements were carried out at 260 nm (Zewail et al., 2019 (link)).
Amount of LEF taken by the cells were quantified using HPLC (Waters 2690 Alliance HPLC system equipped with a Waters 996 photodiode array detector). The Mobile phase consisted of (50%:50%) acetonitrile: phosphate buffer (0.02 M) pH was adjusted to 2.6 with orthophosphoric acid. Flow rate was 1 mL/min and the measurements were carried out at 260 nm (Zewail et al., 2019 (link)).
7
HPLC Analysis of Compound Mixture
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Analysis was performed utilizing HPLC (Waters 2690 Alliance HPLC system coupled with Waters 996 photodiode array detector). Analysis was performed using C18 Inertsil ODS column with dimensions of 4.6 × 250 mm and 5 µm particle size.
8
HPLC Analysis of Compound Standards
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High-performance liquid chromatography (HPLC) analysis was assessed using the Waters 2690 Alliance HPLC system (Milford, CT, USA) equipped with a Waters 996 photodiode array detector. The combined methanolic solution of the nine different standards was prepared, and an amount of 10 µL was injected into the C18 column Inertsil ODS, which had the dimensions of 4.6 × 250 mm and a particle size of 5 µm. The mobile phase consisted of 0.1% phosphoric acid in water: acetonitrile with a constant flow rate of 1 mL/min, and a pH of 3.5 [41 (link)]. The mobile phase consisted of 0.1% phosphoric acid in water: acetonitrile, with a constant flow rate of 1 mL/min and a pH of 3.5 [41 (link)]. Gradient elution was conducted using a start mixture of acidic water: acetonitrile (95:5, v/v for 5 min and then increasing a concentration of acetonitrile to 80% over 30 min followed by a plateau period of 25 min, and finally, the acetonitrile concentration was gradually decreased to 10%, which was then held isocratically for 5 min. The absorbance was measured at 280 nm.
9
HPLC Analysis of Mushroom Extracts and Vitamins
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Solutions (50 mg/mL) of the three mushroom extracts and a standard solution in methanol of three fat vitamins; E, D3, and A, at 806.2, 114, and 400 IU/mL, respectively, were filtered using 0.22 µm syringe filters; then, 10 µL of each was injected onto HPLC column, Waters 2690 Alliance HPLC system equipped with photodiode array detector. Column Inertsil ODS 3: 4.6 × 250 mm, 5 µ, mobile phase: Methanol 100% with isocratic elution of flow rate 1 mL/min at wavelength: 210 nm [27 (link)].
10
Phenolic and Flavonoid Profiling of Propolis
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The qualitative identification of phenolic and flavonoid compounds in Propolis was carried out by HPLC by Nawah Scientific Inc. (Mokatam, Cairo, Egypt). A Waters 2690 Alliance HPLC system equipped with Waters 996 photodiode array detector, Milford, CT, USA. Laminar Flow (model 1386, Thermo Fisher Scientific, Waltham, MA, USA). A stock solution of 10 different standards in methanol was prepared and filtered using a 0.22 μm syringe filter, then 10 μl was injected. 137.8 mg/ml of propolis were accurately weighted and sonicated for 15 min, filtered using a 0.22 μm Nylon syringe filter, then 10 μl was injected. The gradient method that was eventually chosen following a series of preliminary studies uses a mixture of 0.1% Phosphoric acid in water: Acetonitrile (mobile phase). The total runtime of the method was 80 minutes. The absorbance was measured at 280 nm. The total phenolic content (mg/mL) was calculated using the standards; Gallic acid, chlorogenic acid, ellagic acid, and caffeic acid. The total flavonoid content (mg/mL) was calculated using the following standards; catechin, rutin, hesperidin, apigenin, kaempferol, and quercetin.
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