The drug loading was quantified by a Sykam HPLC system with an auto-injector Sykam 5300, a Sykam S3250 UV/Vis detector and the Sykam quaternary pump system 5300 made in Germany, with a Zorbax Rx-Sil column of 4.6×150 mm, and with a 5 μm particle size (Agilent). The standard solutions of PAS ie, 10ppm, 20ppm, 30, pm, 40ppm and 50ppm were prepared from the stock solution of PAS. The empty MgLH (without drug) sample solution was also prepared by dissolving it in a HCl solution for any matrix effect in the HPLC analysis. The mobile phase consisted of a 17.5 mM potassium phosphate buffer as solvent-A with a pH of 3.5 adjusted by phosphoric acid and methanol as solvent B. Before HPLC analysis, the column was saturated with the mobile phase for 30 minutes.
Zorbax rx sil
Manufactured by Agilent Technologies
Sourced in United States, Germany
The Zorbax Rx-Sil is a high-performance liquid chromatography (HPLC) column. It is designed for the separation and analysis of a wide range of compounds. The column features a silica-based stationary phase and is suitable for use in reversed-phase HPLC applications.
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Lab products found in correlation
Agilent 1100 series, by Agilent Technologies (3 mentions)
Agilent 1260 infinity series hplc dad system, by Agilent Technologies (2 mentions)
Agilent 1260 prep pump, by Agilent Technologies (2 mentions)
Agilent 1260 prep als, by Agilent Technologies (2 mentions)
Agilent 1100, by Agilent Technologies (2 mentions)
S 3250, by Sykam (1 mentions)
β carotene, by Merck Group (1 mentions)
1200 hplc system, by Agilent Technologies (1 mentions)
Agilent 1200 hplc system, by Agilent Technologies (1 mentions)
Chem station rev b 02 01 sr1 260, by Agilent Technologies (1 mentions)
G1311a pump, by Agilent Technologies (1 mentions)
Zorbax sb c18, by Agilent Technologies (1 mentions)
1100 hplc system, by Agilent Technologies (1 mentions)
Polystyrene standards, by Merck Group (1 mentions)
Milli q plus, by Merck Group (1 mentions)
2 2 azobisisobutyronitrile, by Merck Group (1 mentions)
Ethylene glycol dimethacrylate egdma, by Merck Group (1 mentions)
Acenaphthene, by Merck Group (1 mentions)
Dichloromethane, by Merck Group (1 mentions)
Tetrahydrofuran, by Merck Group (1 mentions)
17 protocols using zorbax rx sil
1
HPLC Quantification of PAS Loading
2
HPLC Analysis of n-Hexane Extract and Fractions
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The HPLC analysis of n-hexane extract and its fractions (F1 and F2) was performed using an Agilent 1260 infinity series HPLC-DAD system (Agilent Technologies, Waldbronn, Germany) equipped with a binary gradient Agilent 1260 prep pump (G1361A) and an autosampler Agilent 1260 prep ALS (G2260A). Agilent diode array detector 1260 DAD VL (G1315D) was employed for the detection of carotenoids. The separation was performed using an Agilent normal phase (NP) silica column (ZORBAX RX-Sil, 5μm, 4.6 X 150 mm). The following solvents (A) n-hexane and (B) acetone were used at a flow rate of 1 mL/min using a gradient between solvents A and B following the method of Prum et al. [24 (link)] with some modifications as follows: B was run at 0 to 30% for 5 min, 30 to 50% for 15 min, 50 to 100% for 3 min, and maintaining 100% of B until the end of the separation at 30 min. The peaks were integrated at 450 nm. β-carotene (Sigma-Aldrich) was used as a standard to identify the isolated β-carotene.
3
Extraction and HPLC Analysis of Vitamins and Cholesterol
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In the analyses of vitamin E, β-carotene, and cholesterol, the methodology by Prates et al. (32 (link)) was employed, where muscle samples (freeze-dried) underwent direct saponification and single extraction with n-hexane in duplicate. Subsequently, the organic phases were filtered (aliquot from the n-hexane phases) using a 0.45 μm hydrophobic filter and then analyzed by High-Performance Liquid Chromatography (HPLC) in the normal phase. Fluorescence detections were used for tocopherols and tocotrienols, and a UV–Vis photodiode array was used in conjunction for cholesterol and β-carotene. The analysis utilized a normal silica column (Zorbax Rx-Sil, particle diameter of 5 μm, 4.6 mm ID × 25 cm, Agilent Technologies Inc., Santa Clara, CA, United States).
4
HPLC Analysis of Chemical Compounds
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Chromatographic analysis was performed using an Agilent Technologies 1200 HPLC system (Santa Clara, CA, USA) consisting of a binary pump, degasser, a thermostat for the column, and the photodiode array detector. Samples were injected through a Rheodyne injector valve with a 20 μL sample loop. The analytical column Zorbax RX-SIL, 250 mm × 4.6 mm, 5 μm (Agilent Technology, Santa Clara, CA, USA) was used as the stationary phase. The mobile phase consisted of a mixture of acetonitrile and 40 mM ammonium formate buffer pH = 2.8 (80 : 20 v/v). The column temperature (T = 25°C), the flow rate of the mobile phase (F = 1 mL/min), and the wavelength (λ = 255 nm) were kept constant during the analysis.
5
Oxidation Product Separation and Analysis
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Oxidation products were analysed as described (Newie et al. 2017 ) on straight phase (SP) column Zorbax RX‐SIL; (150 × 2.1 mm, particle size 5 μm; Agilent, Waldbronn, Germany) via high pressure liquid chromatography (HPLC) with constant flow rate of 0.2 ml·min−1 solvent system containing n‐hexane/2‐propanol/trifluoroacetic acid (100:1:0.1, v/v/v) coupled to a diode array detector. After collection of the compounds of interest, stereoisomers were separated on chiral phase (CP) column CHIRALCEL OD‐H (150 × 2.1 mm, Agilent) with constant flow rate 0.1 ml·min−1. The solvent system ratio of n‐hexane/2‐propanol/trifluoroacetic acid differed according to the hydroxide to be analysed. The separation of stereoisomers was achieved with a solvent ratio 100:5:0.1 for all HODE and HOTE isomers; 100:2:0.1 (v/v/v) for 8‐HETE, 12‐HETE, 15‐HETE; and 100:1:0.1 (v/v/v) for 5‐HETE and 11‐HETE. All isomers were identified with authentic standards beside the stereoisomers 11R‐hydroxy hexadecatrienoic acid (HHTE)/11S‐HHTE and 11R‐hydroxy hexadecadienoic acid (HHDE)/11S‐HHDE, were tentatively assigned. Absorbance was constantly recorded at wavelength of 234 nm and 220 nm.
6
Simultaneous Analysis of Cholesterol, Carotenoids, and Tocopherols in Meat and Feed
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The simultaneous analysis of total cholesterol, β-carotene, and tocopherols in fresh meat (0.75 g) and feed (0.10 g) was performed according to Prates et al. (2006) . After the direct saponification of samples, an aliquot of the n-hexane layer was filtered and injected into an HPLC system (Agilent 1100 Series; Agilent Technologies Inc., Palo Alto, CA), using a normal-phase silica column (Zorbax RX-Sil, 250 mm × 4.6 mm i.d., 5-μm particle size; Agilent Technologies Inc., Palo Alto, CA), with fluorescence detection of tocopherols and tocotrienols (excitation λ = 295 nm and emission λ = 325 nm) and UV-visible photodiode array detection of cholesterol (λ = 202 nm) and β-carotene (λ = 450 nm) in series. Total cholesterol, β-carotene, tocopherols, and tocotrienols contents were calculated, in duplicate, based on the external standard technique from a standard curve of peak area vs. concentration.
7
Chromatographic Separation and Quantification of Vitamin E Compounds
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Chromatographic separation of α-TCP and tocotrienols was carried out using Agilent HPLC 1200 system (Agilent Technologies, Waldbronn, Germany) with a quaternary pump solvent delivery system, degasser and autosampler at 35°C using a YMC C18 silica gel column (ZORBAX Rx-SIL, 4.6 mm × 250 mm; Agilent Technologies, Palo Alto, CA, USA). The mobile phases used were 99.5% hexane and 0.5% iso-propanol at a flow rate of 1.0 ml/minute. The injection volume was 10 μL. The detection was set at 295 nm excitation and 325 nm emission. Total vitamin E content was quantified using the calibration curves of corresponding standard solution containing α-TCP, α-tocotrienol, β-tocotrienol, γ-tocotrienol and δ-tocotrienol.
8
Simultaneous Quantification of TCHR and Vitamin E
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The simultaneous determination of TCHR and vitamin E (α-tocopherol) contents was performed as previously described [20 (link)]. Briefly, the method involves a direct saponification of the dried salted cod edible portion (500 mg) with a saponification solution (11% w/v potassium hydroxide in a mixture of ethanol: Milli Q water (55:45 v/v)), performed at +80 °C for 15 min, followed by a single n-hexane extraction and HPLC analysis of the extract. Quantification was performed by normal-phase HPLC (column Zorbax Rx Sil, 4.6 mm ID × 250 mm, 5 μm particle size, Agilent Technologies Inc., Palo Alto, CA, USA), with UV–VIS photodiode array (cholesterol) and fluorescence (tocopherols) detectors in tandem. The contents of total cholesterol and α-tocopherol were calculate based on the external standard technique from a standard curve of peak vs. compounds concentrations.
9
Vitamin E Analysis in Rice
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Vitamin E was extracted from rice samples with 2-propanol, and the extract was subjected to liquid chromatography with tandem mass spectrometry (LC-MS/MS) as described previously33 (link). Separation was performed at 40 °C using a silica column (ZORBAX Rx-SIL, 4.6 × 250 mm; Agilent, Palo Alto, CA, USA). A mixture of hexane/1,4-dioxane/2-propanol (100:4:0.5) was used as the mobile phase at a flow rate of 1.0 mL/min. Toc and T3 were detected in atmospheric pressure chemical ionization mode (APCI). MS/MS parameters were optimized with Toc and T3 standards in APCI mode (positive). Toc and T3 were detected using multiple reaction monitoring as follows: α-Toc, m/z 431.3 > m/z 165.1; β-Toc, m/z 417.3 > m/z 151.3; γ-Toc, m/z 417.3 > m/z 151.0; δ-Toc, m/z 403.3 > m/z137.0; α-Toc-3, m/z 425.3 > m/z 165.1; β-Toc-3, m/z 411.3 > m/z 151.1; γ-Toc-3, m/z 411.3 > m/z 151.2; δ-Toc-3, m/z 397.2 > m/z 137.0. Toc and T3 concentrations in the rice samples were calculated using calibration curves for standard Toc and T3 concentrations.
10
SFC-MS Method Development for Compound Analysis
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All SFC-MS experiments were performed on a 1260 Infinity Hybrid SFC/UHPLC analytical system coupled with a 6130 single quadruple MS detector (Agilent Technologies, Santa Clara, CA, USA). The 1260 Infinity Hybrid SFC/UHPLC analytical system consisted of an Infinity SFC binary pump, an Aurora A5 Fusion Module, a degasser, an autosampler with 5 μL loop, a column oven, a make-up flow pump and a backpressure regulator. The operating software was Agilent OpenLab ChemStation Edition C.01.05. All chromatograms were converted into txt. files and then redrawn using Microcal Origin 8.5. Ultrasonication extraction was performed using a KQ-250B ultrasonic water bath (Kunshan Ultrasonic Instrument Co., Ltd, Jiangsu, China).
Ten different columns were tested during method development, namely, ZORBAX RX-SIL, ZORBAX C18 (Agilent Technologies, Santa Clara, USA), Unitary Diol, Unitary NH2, Unitary XAmide (Acchrom Technologies, Beijing, China), Venusil NP, Venusil PFP, Venusil Imidazolyl, Venusil HILIC (Bonna-Agela, Tianjin, China) and Cosmosil 5X-SIL (Nacalai Tesque Inc, Kyoto, Japan). Except for ZORBAX RX-SIL, ZORBAX C18 and Unitary XAmide (150 mm × 4.6 mm, 5 μm), the dimensions of all columns were 250 mm × 4.6 mm with 5 μm particle size.
Ten different columns were tested during method development, namely, ZORBAX RX-SIL, ZORBAX C18 (Agilent Technologies, Santa Clara, USA), Unitary Diol, Unitary NH2, Unitary XAmide (Acchrom Technologies, Beijing, China), Venusil NP, Venusil PFP, Venusil Imidazolyl, Venusil HILIC (Bonna-Agela, Tianjin, China) and Cosmosil 5X-SIL (Nacalai Tesque Inc, Kyoto, Japan). Except for ZORBAX RX-SIL, ZORBAX C18 and Unitary XAmide (150 mm × 4.6 mm, 5 μm), the dimensions of all columns were 250 mm × 4.6 mm with 5 μm particle size.
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