Polar phase compounds were extracted following a previously described method5 (link),48 (link),49 (link) with some modifications. Fifty-mg of ground frozen pepper seeds was mixed with 1.2 mL of methanol and shaken at 75 °C for 30 min; then centrifuged at 13,500×g for 10 min. Next, 0.7 mL of the supernatant was transferred to a 2 mL-microtube and mixed with 0.5 mL of chloroform and 20 μL of ribitol (internal standard). Immediately after adding 0.7 mL of distilled water, it was centrifuged at 2500×g for 10 min. Then 0.5 mL of the supernatant was concentrated using a nitrogen evaporator (MG-2200, Eyela, Japan). Methoxyamine hydrochloride (50 μL) was added and incubated at 37 °C for 2 h. After incubation, 40 μL of sample and 100 μL of N-methyl-N-trifluoroacetamide were mixed and incubated at 37 °C for 30 min. Then, 1 µL of the sample was injected into the GC–MS ISQ LT system (Thermo Fisher Scientific, Waltham, MA, USA) using an auto sampler. The DB-5-fused silica capillary column (0.25 mm × 30 m × 0.25 μm, Agilent, Santa Clara, CA, USA) was used and the oven temperature was set to increase from 50 °C to 310 °C at a rate of 5 °C min−1. The injector was in the split-less mode at 250 °C. Helium was used as the carrier gas at a flow rate of 1 × 10–3 L min−1. The range of mass scan was from 35 to 550 m/z.
Mg 2200
Manufactured by Eyela
Sourced in Japan
The MG-2200 is a laboratory equipment designed for general research and analysis purposes. It is a multi-functional device that can perform various tasks within a controlled laboratory setting. The core function of the MG-2200 is to facilitate data collection and analysis for scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
Isq lt gc ms system, by Thermo Fisher Scientific (1 mentions)
Db 5 fused silica capillary column, by Agilent Technologies (1 mentions)
Hp innowax, by Agilent Technologies (1 mentions)
Agilent 7683, by Agilent Technologies (1 mentions)
Agilent 6890n, by Agilent Technologies (1 mentions)
Isopropanol, by Merck Group (1 mentions)
Sep pak c18 cartridge, by Waters Corporation (1 mentions)
Glycerol, by Merck Group (1 mentions)
Acetone, by Merck Group (1 mentions)
Iodomethane, by Merck Group (1 mentions)
6890 gc 5975 msd, by Agilent Technologies (1 mentions)
4 protocols using mg 2200
1
GC-MS Analysis of Polar Compounds from Pepper Seeds
2
Lipid Extraction and Fatty Acid Analysis of Breast Meat
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lipids were extracted from breast meat using the methods described by Folch et al. (1957) (link). Briefly, 5 g of sample was homogenized with 25 mL of chloroform:methanol (2:1, v/v) at 13,500 rpm using a homogenizer (Ultra Turax T25 basic, Ika WerkeGmbh& Co., Germany). KCl (0.88%) was added to the homogenates, which were then centrifuged at 3,000 rpm for 10 min. The supernatant was evaporated at 38℃ on an N2 gas blow concentrator (MG 2200, Eyela Co., Japan). The methylation was performed according to AOAC (2007) . The fatty acid composition was analyzed in an Agilent 6890 N (Agilent Technologies, USA) equipped with an automatic sampler Agilent 7683 (Agilent Technologies, USA) using an HP-Innowax (30 m length × 0.32 mm i.d. × 0.25 μm film thickness; Agilent Technologies, USA). One microliter of sample was injected (split 1:10; 260℃), and the flow rate was 1.0 mL/min by using helium. The oven temperature was set at 150℃ for 1 min, 150-200℃ at 15℃/min, 200-25℃ at 3℃/min, and 250℃ for 5 min. The detector (FID) was set at 280℃.
3
Plasma Lipid Extraction for LC/MS Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plasma lipids were extracted using the Folch technique [58 (link),59 (link)]. In summary, 20 μL aliquots of plasma were mixed with 780 μL MS grade water (PURELAB Option-Q, UK), followed by 2 mL methanol (A456-4, LC/MS Grade, Thermo Fisher Scientific, Waltham, MA, USA) and 1 mL chloroform (9831-2, LC/MS reagent, J.T.Baker, Phillipsburg, NY, USA) in the ratio of 2:1:0.8 (methanol:chloroform:sample). The mixture was vortexed for 15 s. Then, another 1 mL of MS-grade water was added along with 1 mL of chloroform, and then vortexed. The sample tubes were centrifuged at 800× g, 4 °C for 10 min. The upper aqueous solvent was discarded, and the lower organic phase was collected into a new glass vial, which was then evaporated with a nitrogen gas evaporator (Eyela MG-2200; Japan). It was then solubilized using sample buffer (2:1:1; Isopropanol, LC/MS Grade Merck Millipore, MA, USA; Acetonitrile; H2O, LC/MS reagent, 9831-2, J.T.Baker, Phillipsburg, NY, USA) and stored at −80 °C until LC/MSE analysis.
4
Methylation Analysis of Polysaccharides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Methylation analysis was performed according to the methods of Hakomori (1964) (link) and Kim, Hong, & Shin (2017) (link) with slight modifications. In summary, 20 μL of glycerol (Sigma) was added to 1 mL of polysaccharide sample (1 mg/mL), and the mixture was dried under a nitrogen-flushing with heating block (Eyela MG-2200, Tokyo, Japan). After the sample was methylated by addition of sodium methylsulfinyl carbanion with DMSO and iodomethane (sigma), the methylated sample was collected using an Sep-pak C18 cartridge (Waters, Dublin, Ireland) eluted with EtOH. The methylated product was hydrolyzed with 1 M TFA for 90 min at 121 °C, reduced with NaBD4 for 150 min, and finally acetylated with acetic anhydride for 150 min at 100 °C. The resulting methylated alditol acetates (PMAAs) were dissolved with 50 μL of acetone (Sigma) and analyzed using a GC–MS (6890 GC/5975 MSD; Agilent, Santa Clara, CA, USA) equipped with a SP-2380 capillary column. The analysis was performed with the following temperature program: 60 °C (1 min) → 150 °C (30∘C/min) → 180 °C (1 °C/min) → 231 °C (1.5 °C/min) → 250 °C (30 °C/min), 250 °C (10 min). PMAAs were identified by their fragment ions and relative retention times, and their mole percentage was estimated from the peak areas and response factors on a flame ionization detector.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!