The determination of α-, β-, γ-, and δ-tocopherol was accomplished by following the UNI EN 12822:2014 p.to 5.4 protocol [25 ]. Briefly, an exactly weighted amount of about 2.0 g of olive oil was solubilized in 8 mL of acetone and injected (injection volume 5 µL) into a ZORBAX® Eclipse XBD-C18, 4.6 × 150 mm, 5 µm analytical column, eluted with MeOH (0.1% acetic acid v/v)/AcOEt (100:0 for 3.5 min, 100:0 → 10:90 over 0.7 min, 10:90 for 2 min). The tocopherol content, expressed in mg/kg, was calculated by measuring the areas of the related chromatographic peaks.
Eclipse xbd c18
Manufactured by Agilent Technologies
Sourced in United States
The Eclipse XBD-C18 is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of compounds. It features a C18 stationary phase, which is a commonly used reversed-phase material for the separation of both polar and non-polar analytes. The column is designed to provide efficient, high-resolution separations with good peak shape and reproducibility.
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Lab products found in correlation
6520 accurate mass q tof instrument, by Agilent Technologies (2 mentions)
500 mhz spectrometer, by Bruker (2 mentions)
Ultrospec 2100 uv visible spectrophotometer, by Harvard Bioscience (1 mentions)
1260 infinity system, by Agilent Technologies (1 mentions)
Zorbax eclipse plus c18, by Agilent Technologies (1 mentions)
1100 lc msd, by Agilent Technologies (1 mentions)
Xbridge peptide beh c18, by Waters Corporation (1 mentions)
Dextran standard, by Merck Group (1 mentions)
Waters 2414 differential refractive index detector, by Waters Corporation (1 mentions)
G4000swxl column, by Tosoh (1 mentions)
Zorbax cartridge guard column, by Agilent Technologies (1 mentions)
12 protocols using eclipse xbd c18
1
Olive Oil Tocopherol Determination
2
LC-MS/MS Analysis of Small Molecules
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LC-MS/MS analysis was performed using a Zorbax Eclipse XBD-C18, 4.6 × 250 mm (5 μm) and the mobile phase consisted of 0.5% formic acid in Milli-Q water (eluent A) and acetonitrile + 0.5% formic acid (eluent B). A flow rate of 0.3 ml/min was used, with the following gradient program: 0–30 min from 70 to 5% A, 30–45 min at 5% A, 45–65 min 70% A.
3
HPLC Analysis of A. precatorius Seeds
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Excellent separations and suitable retention time of A. precatorius seeds were obtained in isocratic elution. Separation was carried on ZORBAX Eclipse XBD-C 18 (4.6 x 150 mm), 5 μm particle size column. Phosphate buffer:acetonitrile (80:20 v/v) was used as a mobile phase at a flow rate of 0.9 mL/min. The detection was carried out by using DAD at 220 nm. 10 μL of the test solution was injected in to the HPLC system. The column temperature was kept at 40°C.
4
Quantifying Ozone and Microplastics
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Ultrospec 2100 UV-Visible Spectrophotometer (Biochrom, USA) was used to measure the UV absorption and standardize the O3 stock solutions. The Indigo method [Bader and Hoigné, 1981] was also employed for dissolved O3 measurements. MPs were detected using an Infinity Series HPLC of Agilent coupled with UV/Vis detector. A Zorbax Eclipse XBD C18 (150 × 4.6 mm i.d; 5 μm particle size) was the employed column.
Acetonitrile (A) and Milli-Q water adjusted to pH = 3 by orthophosphoric acid (B) were employed as mobile phases. The analyses were performed under a gradient method as follows: 30% A and 70% B initially kept for 5 min, 30% A to 60% A during 5 min, 60%
A and 40% B kept for 25 min, 60% A to 80% A during 5 min, 80% A and 20% B kept for 30 min, 80% A to 30% A during 10 min and finally 30% A and 70% B kept for 10 min. Total organic carbon (TOC) was determined using a TOC-V CNS Total Organic Carbon analyzer by Shimadzu (Japan).
Acetonitrile (A) and Milli-Q water adjusted to pH = 3 by orthophosphoric acid (B) were employed as mobile phases. The analyses were performed under a gradient method as follows: 30% A and 70% B initially kept for 5 min, 30% A to 60% A during 5 min, 60%
A and 40% B kept for 25 min, 60% A to 80% A during 5 min, 80% A and 20% B kept for 30 min, 80% A to 30% A during 10 min and finally 30% A and 70% B kept for 10 min. Total organic carbon (TOC) was determined using a TOC-V CNS Total Organic Carbon analyzer by Shimadzu (Japan).
5
Quantification of Lutein and Esters
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The lutein and hydrolysed lutein esters’ contents were determined using high-performance liquid chromatography (Infinity 1260 system; Agilent Technologies, Santa Clara, CA, USA), with a C18 precolumn (Eclipse XBD-C18; 4.6 × 12.5 mm; ID, 5 μm) and column (Zorbax Eclipse Plus C18; 4.6 × 150 mm; ID, 3.5 μm) (both from Agilent Technologies). The mobile phase was acetonitrile:methanol (90:10; v/v), with an injection volume of 10 µL. The elution was isocratic over 15 min at a flow rate of 0.8 mL/min. The analysis was performed at 30 °C (sample temperature, 20 °C) and the eluted components were determined with a UV-Vis diode array detector, at 446 nm. Lutein was identified and quantified according to retention time and absorption spectra, compared to the lutein standard. All of the analyses were performed in triplicate. The procedures for the calibration curves and sample preparation are described in the Supplementary Materials .
6
Quantifying Encapsulation and Loading of Micelles
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The Cur-MMs sample (or C6-MMs) (0.1 mL) was dissolved in 10 mL of methanol. In order to completely break up the micelles, the mixture was placed in an ultrasonic bath for 10 min. The solution was filtered through 0.22 μm membrane filters and subjected to high-performance liquid chromatography (HPLC) using an Agilent Eclipse XBD C18 reverse-phase column (4.6 mm × 250 mm, 5 μm). The mobile phase consisted of a mixture of acetonitrile and water (52:48, v/v). The flow rate was 1.0 mL/min at 25 °C, and the detection wavelength was 430 nm. The DL and EE were calculated as shown below Equations (1) and (2) [27 (link),28 (link)]:
7
Synthesis and Characterization of β-Diketoesters
8
Rutin Quantification by HPLC
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The rutin content of the extract was determined chromatographically using HPLC system [19 (link), 20 (link)] of Agilent technologies, with column from Agilent eclipse XBD® C 18 bonded with 5 µm (4.6 × 150 mm). Before starting validation, system suitability parameter was calculated. It was determined by taking percent relative standard deviation (RSD) of the five standards injections using the same concentration of rutin by HPLC method. The precision of system was checked as per the developed method by using multiple injections of a homogeneous standard solution. This indicated the performance of the HPLC instrument under the chromatographic condition. As a part of method validation minimum five injections of the standard preparation were performed for inter day precision. The relative standard deviation was not more than 2.0%. Limits of detection (LOD) and Limit of quantification (LOQ) were calculated by method based on standard deviation (σ) and slope (S) of calibration plot using formula LOD = 3.3 σ/S and LOQ = 10 σ/S.
9
Azide-Modified Isoprenoid Tagging and Analysis
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Azide-modified isoprenoids
(neryl, geranyl, or BP/BPP) were labeled with one equivalent of TAMRA-DBCO
(100 μM) in either water or UppS/potato acid phosphatase reaction
conditions directly. Reactions were typically complete in under 60
min. Products were analyzed after this time on an Agilent 1100 HPLC
system (Agilent Eclipse XBD-C18, 3.5 μM, 4.6 × 50 mm) monitoring
for the TAMRA fluorophore (454/525 ex/em). A gradient method was used
to separate BPPs and BPs with 100 mM ammonium bicarbonate (A) and n-propanol (B). Line B was increased from 15 to 95% over
36.9 min and then held at 95% until 42 min. LC–MS analysis
of non-conjugated azide materials was performed on an Agilent 1260
LC and 6000 series ESI-MS single quad with four channels for monitoring
selected ions (Waters XBridge Peptide BEH C18, 3.5 μM, 4.6 ×
50 mm). n-Propanol was increased at a rate of 4%
per min, starting at 20% with 80% of a 0.1% ammonium hydroxide solution
as the aqueous component. Mass values for Az-BPs (1Z-10Z, where Z is the number of Z-configuration isoprene
additions) were scanned following potato acid phosphatase treatment.
(neryl, geranyl, or BP/BPP) were labeled with one equivalent of TAMRA-DBCO
(100 μM) in either water or UppS/potato acid phosphatase reaction
conditions directly. Reactions were typically complete in under 60
min. Products were analyzed after this time on an Agilent 1100 HPLC
system (Agilent Eclipse XBD-C18, 3.5 μM, 4.6 × 50 mm) monitoring
for the TAMRA fluorophore (454/525 ex/em). A gradient method was used
to separate BPPs and BPs with 100 mM ammonium bicarbonate (A) and n-propanol (B). Line B was increased from 15 to 95% over
36.9 min and then held at 95% until 42 min. LC–MS analysis
of non-conjugated azide materials was performed on an Agilent 1260
LC and 6000 series ESI-MS single quad with four channels for monitoring
selected ions (Waters XBridge Peptide BEH C18, 3.5 μM, 4.6 ×
50 mm). n-Propanol was increased at a rate of 4%
per min, starting at 20% with 80% of a 0.1% ammonium hydroxide solution
as the aqueous component. Mass values for Az-BPs (1Z-10Z, where Z is the number of Z-configuration isoprene
additions) were scanned following potato acid phosphatase treatment.
10
Monosaccharide Composition and Relative Mw of Polysaccharides
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The precolumn derivatization approach was used to determine the monosaccharide composition of polysaccharides [17 (link)]. The sample was dissolved by adding 3 mg of polysaccharide to 1 mL of distilled water and hydrolyzed with 1 mL (4 mol/L) of trifluoroacetic acid (TFA) at 110 °C for 6 h. The residual TFA was then removed by adding methanol. 1 mL of hydrolysis product was added to distilled water for derivatization. After derivatization, the derivatives filtered through a 0.22-μm membrane were detected by an Agilent Eclipse XBD-C18.
The relative Mw of polysaccharides was measured using a high-performance liquid chromatography system equipped with a Waters-2414 differential refractive index detector (Waters Co., Milford, MA, USA). A G4000SWXL column (7.8 × 300 mm, Tosoh Co., Ltd. Tokyo, Japan) was used. The column temperature was 40 °C, and 0.1 M sodium nitrate solution was used as the eluent. The relative Mw of polysaccharides was calculated according to calibration curves (Log Mol Wt = 1.27e − 8.10e−1 T, R2 = 0.9943) of the dextran standards (Sigma Co., St Louis, MI, USA), including 667,000 Da, 413,000 Da, 76,900 Da, 43,500 Da, 10,500 Da, and 5000 Da.
The relative Mw of polysaccharides was measured using a high-performance liquid chromatography system equipped with a Waters-2414 differential refractive index detector (Waters Co., Milford, MA, USA). A G4000SWXL column (7.8 × 300 mm, Tosoh Co., Ltd. Tokyo, Japan) was used. The column temperature was 40 °C, and 0.1 M sodium nitrate solution was used as the eluent. The relative Mw of polysaccharides was calculated according to calibration curves (Log Mol Wt = 1.27e − 8.10e−1 T, R2 = 0.9943) of the dextran standards (Sigma Co., St Louis, MI, USA), including 667,000 Da, 413,000 Da, 76,900 Da, 43,500 Da, 10,500 Da, and 5000 Da.
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