Materials used for incubations included: pooled human liver microsomes and sodium salt of NADPH (Sigma-Aldrich, Saint-Louis, MO, USA), monopotassium phosphate and dipotassium phosphate (POCH, Gliwice, Poland), and deionized water, 18 MΩ resistance (Millipore, Bellerica, MA, USA). Dimethyl sulfoxide (Sigma-Aldrich) was also used to prepare stock solutions of compounds 5 and 6. The LC-MS internal standard was a 5 μM buspirone hydrochloride solution (Sigma-Aldrich). The mobile phase consisted of deionized water, HyperGrade LC-MS acetonitrile (MERCK, Darmstadt, Germany) and formic acid for LC-MS (Sigma-Aldrich).
Hypergrade lc ms acetonitrile
Manufactured by Merck Group
Sourced in Germany
HyperGrade LC-MS acetonitrile is a high-purity solvent used in liquid chromatography-mass spectrometry (LC-MS) applications. It is a colorless, volatile liquid that meets the specifications required for use in sensitive analytical techniques.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Formic acid for lc ms, by Merck Group (1 mentions)
Pooled human liver microsomes, by Merck Group (1 mentions)
Milli q purification system, by Merck Group (1 mentions)
Hplc grade formic acid, by Dikma Technologies (1 mentions)
Taxifolin, by Merck Group (1 mentions)
Procyanidin b2, by Merck Group (1 mentions)
D salicin, by Merck Group (1 mentions)
D quinic acid, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Gallic acid, by Merck Group (1 mentions)
Folin ciocalteu reagent, by Merck Group (1 mentions)
4 protocols using hypergrade lc ms acetonitrile
1
Quantitative LC-MS Analysis of Compounds
2
CYP Metabolizing Enzyme Kinetics
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Incubations for determination of leading metabolizing CYP were conducted using P450 isolated isoenzymes in the form of cytochrome P450 BACULOSOMES AE Plus Reagents (Thermo Fisher Scientific, USA) with the addition of sodium salt of NADPH (Sigma-Aldrich, USA), monopotassium phosphate and dipotassium phosphate (POCH, Poland), and deionized water (18 MΩ, Merck-Millipore, Germany). Dimethyl sulfoxide (Sigma-Aldrich, USA) was used to prepare stock solutions of the studied compounds.
Incubations for IC 50 value estimation were conducted using pooled human liver microsomes (Sigma-Aldrich, St. Louis, MO, USA). Testosterone, as well as 6-OH-testosterone, were purchased from Sigma-Aldrich, USA.
The mobile phase of the LC separation consisted of deionized water and Hypergrade LC-MS acetonitrile (Merck, Germany). Both mobile phases were prepared with the addition of ammonium formate buffer, which consisted of LC-MS formic acid and ammonium formate. The LC-MS internal standard was a 5 µM buspirone hydrochloride solution.
Incubations for IC 50 value estimation were conducted using pooled human liver microsomes (Sigma-Aldrich, St. Louis, MO, USA). Testosterone, as well as 6-OH-testosterone, were purchased from Sigma-Aldrich, USA.
The mobile phase of the LC separation consisted of deionized water and Hypergrade LC-MS acetonitrile (Merck, Germany). Both mobile phases were prepared with the addition of ammonium formate buffer, which consisted of LC-MS formic acid and ammonium formate. The LC-MS internal standard was a 5 µM buspirone hydrochloride solution.
3
Synthesis and Characterization of Peptides
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The synthetic peptides CTP
(YGRRARRRRRR) and GABA–CTP were synthetized by SBS Genetech
Co., Ltd. in a purity of 98%. The molecular weights of the peptides
were confirmed by mass spectrometry. The lyophilized peptides were
prepared and diluted in 0.9% NaCl before use.
GABA was purchased
from Sigma Aldrich (St. Louis, MO), with purity ≥99%.
LC–MS hypergrade acetonitrile was purchased from Merck (Darmstadt,
Germany). High-performance liquid chromatography (HPLC)-grade formic
acid was purchased from Dikma Technologies Inc. (Lake Forest, CA).
HPLC-grade ammonium formate was purchased from Sinopharm Chemical
reagent Co., Ltd (Shanghai, China). All other chemicals and reagents
of reagent grade were purchased from Sigma Aldrich (St. Louis, MO).
Ultrapure water was prepared using a Milli-Q purification system (Millipore,
Bedford, MA).
(YGRRARRRRRR) and GABA–CTP were synthetized by SBS Genetech
Co., Ltd. in a purity of 98%. The molecular weights of the peptides
were confirmed by mass spectrometry. The lyophilized peptides were
prepared and diluted in 0.9% NaCl before use.
GABA was purchased
from Sigma Aldrich (St. Louis, MO), with purity ≥99%.
LC–MS hypergrade acetonitrile was purchased from Merck (Darmstadt,
Germany). High-performance liquid chromatography (HPLC)-grade formic
acid was purchased from Dikma Technologies Inc. (Lake Forest, CA).
HPLC-grade ammonium formate was purchased from Sinopharm Chemical
reagent Co., Ltd (Shanghai, China). All other chemicals and reagents
of reagent grade were purchased from Sigma Aldrich (St. Louis, MO).
Ultrapure water was prepared using a Milli-Q purification system (Millipore,
Bedford, MA).
4
Quantification of Polyphenolic Compounds
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The D-(-)-salicin (≥98.5% HPLC), D-(-)-quinic acid (≥98%), Procyanidin B2 (≥90% HPLC) and Taxifolin (≥85% HPLC) analytical standards were purchased from Sigma-Aldrich (Steinheim, Germany). The LC-MS hypergrade acetonitrile was purchased from Merck (LiChrosolv, Germany), and the formic acid (HiperSOLV Chromanorm) was purchased from VWR Chemicals. Milli-Q Type 1 ultrapure water (suitable for chromatography and other advanced analytical techniques) was used for sample preparation as well as the mobile phase. The Folin–Ciocalteu reagent, gallic acid, methanol and other chemicals used for the analyses were of analytical grade and were purchased from Sigma-Aldrich (Steinheim, Germany).
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