The α-tocopherol content was evaluated using high-performance liquid chromatography (HPLC) on a Pinnacle II Silica column (Restek, USA; 5-μm particle size; 150 mm × 4.6 mm), as described previously (Fernandez-Orozco et al., 2003 (link)). Tocopherols were extracted from fresh plant tissues using pure hexane (1:10) by centrifugation at 349 × g for 5 min. The supernatant was filtered through a 0.45-μm polytetrafluoroethylene (PTFE) membrane syringe filter (VWR International, USA). The HPLC 10A system, equipped with an RF-10A fluorescence detector (Shimadzu, Japan), was used for analysis. Peaks were detected at an excitation wavelength of 295 nm and an emission wavelength of 330 nm. The mobile phase (0.5% isopropanol in hexane) was used at a flow rate of 1 ml min−1. The α-tocopherol was identified according to the analytical standard. The α-tocopherol content is expressed per gram dry weight of microgreens.
10a hplc system
Manufactured by Shimadzu
Sourced in Japan
The Shimadzu 10A HPLC system is a high-performance liquid chromatography instrument designed for analytical separations. It features a quaternary solvent delivery system, an autosampler, a column oven, and a variety of detectors. The 10A HPLC system is capable of performing a range of analytical techniques, including reversed-phase, normal-phase, and ion-exchange chromatography.
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Lab products found in correlation
Sil 20ac ht, by Thermo Fisher Scientific (6 mentions)
Xcalibur, by Thermo Fisher Scientific (4 mentions)
Rf 10a fluorescence detector, by Shimadzu (2 mentions)
Carotenoid column, by YMC (2 mentions)
Xcalibur software version 4, by Thermo Fisher Scientific (2 mentions)
Tsq quantum ultra, by Thermo Fisher Scientific (2 mentions)
Lc solution, by Shimadzu (2 mentions)
Ptfe membrane syringe filter, by Avantor (1 mentions)
Spd m10avp, by Shimadzu (1 mentions)
Tsq quantum ultra triple, by Thermo Fisher Scientific (1 mentions)
Kjeltec system 1002 apparatus, by Foss (1 mentions)
Tsq vantage, by Thermo Fisher Scientific (1 mentions)
Accela uplc, by Thermo Fisher Scientific (1 mentions)
Hypersil ods c18 column, by Thermo Fisher Scientific (1 mentions)
Omni bead ruptor 24, by Omni International (1 mentions)
Chaps, by Merck Group (1 mentions)
Tsq quantum ultra triple quadrupole mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Avance 2, by Bruker (1 mentions)
Exactive mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Quattro micro api mass spectrometer, by Waters Corporation (1 mentions)
16 protocols using 10a hplc system
1
Quantifying α-Tocopherol in Microgreens
2
Quantifying Carotenoids in Microgreens
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The contents of lutein-zeaxanthin and β-carotene were evaluated using HPLC on a YMC carotenoid column (YMC, Japan; 3-µm particle size; 150 mm × 4.0 mm), as described previously (Edelenbos et al., 2001 (link)). Plant tissue (1 g) was ground in liquid N2. Then, 10 ml of 80% acetone was added to the sample and mixed. The sample was centrifuged at 5,000 × g for 15 min. The supernatant was filtered through a 0.22-µm nylon membrane syringe filter (VWR International, USA). The HPLC 10A system (Shimadzu, Japan), equipped with a diode array (SPD-M 10A VP) detector, was used for the analysis. Peaks were detected at 440 nm. The mobile phase consisted of solvent A (80% methanol and 20% water) and solvent B (100% ethyl acetate). Carotenoids were identified according to the standards. Carotenoid contents are expressed per gram dry weight of microgreens by the calculation of the ratio of fresh weight to dry weight.
3
Carotenoid Quantification by HPLC
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Contents of lutein and β-carotene were evaluated using HPLC with a diode array detector (at 440 nm), on a YMC Carotenoid column (3 μm particle size, 150 x 4.0 mm; YMC, Japan). Carotenoids were extracted using 80% acetone (1 g of sample grounded with liquid nitrogen 10 ml-1 of solvent), centrifuged (5 min, 349 x g), and filtrated through a 0.45-μm nylon membrane syringe filter (VWR International, USA). The HPLC 10A system (Shimadzu, Japan) equipped with a diode array (SPD-M 10A VP) detector was used for analysis. Peaks were detected at 440 nm. The mobile phase consisted of A (80% methanol, 20% water) and B (100% ethylacetate). Gradient: 0 min; 20% B, 2.5 min; 22.5% B, 20–22.5 min; 50% B, 24–26 min; 80% B, 31–34 min; 100% B, 42–47 min; and 20% B, flow rate 1 ml min-1. The sensitivity of all chromatographic methods was established using a method validation procedure outlined by Edelenbos et al. [24 (link)].
4
Quantifying Alpha Tocopherol Content
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Alpha tocopherol (α-T) content was evaluated according to Fernandez-Orozco et al. [23 (link)] using high-performance liquid chromatography (HPLC) on a Pinacle II silica column, 5 μm particle size, 150 x 4.6 mm (Restek, USA). Tocopherol homologues were extracted using pure hexane (1g of sample / 10 ml of solvent), centrifuged (5 min, 349 x g) and filtrated through 0.45 μm PTFE membrane using s syringe filter (VWR International, USA). The HPLC 10A system, equipped with RF-10A fluorescence detector (Shimadzu, Japan) was used for analysis. Peak was detected using an excitation wavelength of 295 nm and emission wavelength at 330 nm. The mobile phase was 0.5% isopropanol in hexane, flow rate 1 ml min-1.
5
Quantifying Chlorophylls and Carotenoids in Lettuce
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Contents of chlorophylls (a, b) and carotenoids (neoxanthin, violaxanthin, lutein and zeaxanthin, αand β-carotenes) were evaluated according to the methods of Edelenbos (2001) using high-performance liquid chromatography (HPLC) on a Chromegabond C30 column (3 µm particle size, 15 × 2.1 mm) (ES Industries, West Berlin, NJ, USA) [20] (link). Carotenoids were extracted using 80% acetone (500 mg of sample grounded with 10 mL liquid N 2 ), centrifuged (10 min, 4000 rpm min -1 ), and filtrated through a 13 mm and 0.22 µm nylon syringe filter (BGB Analytik AG, Böckten, Switzerland). The HPLC 10A system (Shimadzu, Kyoto, Japan) equipped with a diode array (SPD-M 10A VP) detector was used for analysis. Peaks were detected at 440 nm. The mobile phase consisted of A (80% methanol, 20% water) and B (100% ethyl acetate). Gradient: 0 min; 20% B, 2.5 min; 22.5% B, 20-22.5 min; 50% B, 24-26 min; 80% B, 31-34 min; 100% B, 42-47 min; and 20% B, flow rate 1 mL min -1 . The contents of chlorophylls and carotenoids in each group of lettuce were evaluated after 18, 36, 60, and 84 h after inoculation. The results are expressed as mg g -1 in the dry weight of plants.
6
Lipid Quantification by HPLC-MS/MS
7
Cholesterol Quantification in Lysosomes
8
Quantification of Diacylglycerol Lipids by LC-MS
9
Proximate Composition and Cholesterol Analysis
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The dry matter content was determined [34 ] by drying samples in an oven at 105 °C until a constant weight was obtained (method No. 950.46B) in [34 ]. The crude protein content was determined by the Kjeldahl method using the Kjeltec system 1002 apparatus (Foss-Tecator AB, Höganäs, Sweden), and a conversion factor of 6.25 was used to convert total nitrogen to crude protein (method No. 981.10) in [34 ]. Crude fat was determined by the Soxhlet extraction method (method No. 960.39) in [34 ]. Ash was determined by incineration in a muffle furnace at 550 °C for 24 h (method No. 920.153) in [34 ]. The content of protein, fat and ash was expressed as the weight percentage of dry matter from muscle tissues. The hydroxyproline content was determined by the NMKL-AOAC method [35 (link)].
The cholesterol content in meat was determined according to the extraction method described by Polak et al. [36 (link)], followed by HPLC separation and analysis on a Shimadzu 10 A HPLC system (Shimadzu Corp., Kyoto, Japan). The data collection and evaluation were performed by using the LC Solution (Shimadzu Corp., Kyoto, Japan) operating system. The analytical column was LiChrospher 100 RP-18e, 150 × 4.6 mm, 5 μm (Alltech Associates Inc., Chicago, IL, USA) with a guard column (LiChrospher 100 RP-18, 7.5 × 4.6 mm). The cholesterol content was expressed as mg/100 g of fresh meat.
The cholesterol content in meat was determined according to the extraction method described by Polak et al. [36 (link)], followed by HPLC separation and analysis on a Shimadzu 10 A HPLC system (Shimadzu Corp., Kyoto, Japan). The data collection and evaluation were performed by using the LC Solution (Shimadzu Corp., Kyoto, Japan) operating system. The analytical column was LiChrospher 100 RP-18e, 150 × 4.6 mm, 5 μm (Alltech Associates Inc., Chicago, IL, USA) with a guard column (LiChrospher 100 RP-18, 7.5 × 4.6 mm). The cholesterol content was expressed as mg/100 g of fresh meat.
10
Quantification of Lipid Profiles in Mouse Hearts
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The abundance of PA, DAG, triglyceride, and CL in mouse hearts was determined as described with modification (22 (link)). The LC-MS analysis was performed either with a Shimadzu 10A HPLC system and a Shimadzu SIL-20AC HT auto-sampler coupled to a Thermo Scientific TSQ Quantum Ultra triple quadrupole (TQ) mass spectrometer operated in selected reaction monitoring mode or with a Thermo Fisher Scientific Vantage TSQ mass spectrometer with Thermo Accela UPLC operated by Xcalibur software using selected ion monitoring mode. PA-(14:0)2, DAG-(15:0)2, triglyceride-(17:0)3, and CL-(14:0-14:0)2 were used as internal standards. Quantification of lipids was based on the ratio of the peak area of the analyte to the internal standard. For example, the ratio of CL-(18:2/18:2)2 and CL-(14:0-14:0)2 is used for measurement of CL-(18:2/18:2)2.
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