High-performance liquid chromatography (HPLC)-grade methanol, chloroform, and water were purchased from Fisher Scientific (Pittsburg, PA). HPLC-grade hexane was purchased from Honeywell Burdick & Jackson (Muskegon, MI). Butylated hydroxytoluene (BHT), myristic-d27 acid, methoxylamine hydrochloride, pyridine, and ammonium acetate were purchased from Sigma-Aldrich (St. Louis, MO). BSTFA [N,O-bis(trimethylsilyl) trifluoroacetamide] containing 1% trimethylchlorosilane (TMCS) was purchased from Alfa Aesar (Ward Hill, MA).
Trimethylchlorosilane tmcs
Manufactured by Thermo Fisher Scientific
Sourced in United States
Trimethylchlorosilane (TMCS) is a chemical compound used as a reagent in various laboratory applications. It is a colorless liquid with a pungent odor. TMCS is commonly employed as a silylating agent in organic synthesis and analytical chemistry procedures.
Automatically generated - may contain errors
Lab products found in correlation
Pyridine, by Merck Group (7 mentions)
Methanol, by Merck Group (5 mentions)
1 2 3 13c3 myristic acid, by Cambridge Isotopes (4 mentions)
N methyl n trimethylsilyltrifluoroacetamide mstfa, by Thermo Fisher Scientific (4 mentions)
Acetonitrile, by Merck Group (3 mentions)
13c5 proline, by Cambridge Isotopes (3 mentions)
2h4 succinic acid, by Cambridge Isotopes (3 mentions)
2h7 cholesterol, by Cambridge Isotopes (3 mentions)
Methoxamine hydrochloride, by Merck Group (3 mentions)
N heptane, by Merck Group (3 mentions)
Myristic d27 acid, by Merck Group (2 mentions)
Ammonium acetate, by Merck Group (2 mentions)
Methoxyamine hydrochloride, by Merck Group (2 mentions)
Pyridine, by Thermo Fisher Scientific (2 mentions)
13c5 15n glutamic acid, by Cambridge Isotopes (2 mentions)
13c4 disodium α ketoglutarate, by Cambridge Isotopes (2 mentions)
N methyl trimethylsilyltrifluoroacetamide mstfa, by Thermo Fisher Scientific (2 mentions)
5 13c glutamine, by Cambridge Isotopes (2 mentions)
Myristic 1 2 13c2 acid, by Merck Group (2 mentions)
Milli q system, by Merck Group (2 mentions)
13 protocols using trimethylchlorosilane tmcs
1
HPLC-based Metabolite Profiling
2
Optimized Metabolite Extraction Protocol
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High-performance liquid chromatography-grade chloroform, methanol, and water were purchased from Fisher Scientific (Pittsburg, PA). Ammonium acetate, butylated hydroxytoluene, COR, 2-DG, dimethyl sulfoxide (DMSO), MJ, methoxyamine hydrochloride, myristic-d27 acid, and pyridine were purchased from Sigma-Aldrich (St. Louis, MO). N,O-Bis(trimethylsilyl)trifluoroacetamide (BSTFA) containing 1% trimethylchlorosilane (TMCS) was purchased from Alfa Aesar (Ward Hill, MA), and 1,2-diheptadecanoyl-sn-glycero-3-phosphoethanolamine was purchased from Avanti Polar Lipids (Alabaster, AL).
3
Bisphenol A Quantification Protocol
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BPA (purity N 99%) was supplied by Sigma-Aldrich (St. Louis MO, USA). The internal standard (phenanthrene-d 10 , GC-MS analysis) obtained from Dr. Ehrenstorfer GmbH (Augsburg, Germany). The N,O-bis (trimethylsilyl) trifluoroacetamide (BSTFA, purity N 98%) and trimethylchlorosilane (TMCS, purity N 98%) were purchased from Alfa Aesar (Ward Hill, MA). HPLC grade solvents including methanol, acetonitrile and dichloromethane were purchased from Merck (Germany). The sodium hydrogen phosphate, anhydrous sodium sulfate, sodium hydroxide, hydrochloric acid, and sodium nitrate were all of analytical reagents. The anhydrous sodium sulfate was put into in a muffle furnace and dried at 450 °C for 4 h before usage. To prepare the stock solutions of BPA (1 g L -1 ), an appropriate amount of BPA was dissolved in methanol in a volumetric flask and stored at -20 °C.
4
Stable Isotope-Labeled Compounds for Analytical Standards
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The chemicals used for sample preparation were all of analytical grade, except as otherwise stated. The following were purchased from Cambridge Isotope Laboratories, (Andover, MA, USA): stable isotope labelled IS compound 13C5-proline, 2H4-succinic acid, 13C5, 15 N-glutamic acid, 1,2,3-13C3 myristic acid, 2H7-cholesterol and 13C4 disodium alpha-ketoglutarate. The following were purchased from Campro (Veenendaal, The Netherlands): 13C12-sucrose, 13C4-palmitic acid and 2H4-butanediamine. 2HCl and 13C6-glucose from Aldrich (Steinheim, Germany) and 2H6-salicylic acid from Icon (Summit, NJ, USA). Stock solutions of the IS were prepared either in purified and deionised water (Milli-Q, Millipore, Billerica, MA, USA) or in methanol (J.T.Baker, Deventer, Holland) at the same concentration, 0.5 μg/μl. Metyl stearate was purchased from Sigma (St. Louis, MO, USA). N-Metyl-N-trimethylsilyltriflouroacetamide (MSTFA) with 1% trimethylchlorosilane (TMCS) and pyridine (silylation grade) were purchased from Pierce Chemical Co, Dallas,TX, USA). Heptane was purchased from Fisher Scientific (Loughborough, UK).
5
Isotope-Labeled Metabolite Extraction
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All chemicals were of analytical grade. The isotopically labeled internal standards (IS) [1,2,3-13C3]-myristic acid and [2H6]-salicylic acid were purchased from Cambridge Isotope Laboratories (Andover, MA, USA) and Icon (Summit, NJ, USA) respectively. The stock solutions for IS were prepared in 0.5 μg/μl concentrations in methanol prior to metabolite extraction. Silylation grade pyridine and N-Methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) with 1% trimethylchlorosilane (TMCS) were purchased from Pierce Chemical Co (Rockford, IL, USA).
6
Metabolomic Analysis with Stable Isotopes
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All standard reagents were of analytical grade or equivalent and obtained from Sigma-Aldrich (St Louis, MO, USA), Merck (Darmstadt, Germany) and J.T. Baker (Phillipsburg, NJ, USA). N-Methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) plus 1% trimethylchlorosilane (TMCS) and pyridine were obtained from Thermo Fisher Scientific (Rockford, IL, USA). The stable isotope-labelled internal standards, [13C5]-proline, [2H4]-succinic acid, [13C5,15N]-glutamic acid, [1,2,3-13C3]-myristic acid, [2H7]-cholesterol and [13C4]-disodium α-ketoglutarate were purchased from Cambridge Isotope Laboratories (Andover, MA); [13C6]-glucose, [13C12]-sucrose, [13C4]-hexadecanoic acid and [2H4]-putrescine were purchased from Campro (Veenendaal, The Netherlands) and [2H6]-salicylic acid was obtained from Icon (Summit, NJ). Stock solutions of each internal standard were prepared in either Milli-Q water or methanol to a concentration of 0.5 μg/μL and stored at 4°C.
7
Quantitative Metabolomics: Isotope Standards
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All standard reagents were of analytical grade or equivalent and obtained from Sigma-Aldrich (St Louis, MO, USA), Merck (Darmstadt, Germany) and J.T. Baker (Phillipsburg, NJ, USA). N-Methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) plus 1% trimethylchlorosilane (TMCS) and pyridine were obtained from Thermo Fisher Scientific (Rockford, IL, USA). The stable isotope-labelled internal standards, [13C5]-proline, [2H4]-succinic acid, [13C5,15N]-glutamic acid, [1,2,3-13C3]-myristic acid, [2H7]-cholesterol and [13C4]-disodium α-ketoglutarate were purchased from Cambridge Isotope Laboratories (Andover, MA); [13C6]-glucose, [13C12]-sucrose, [13C4]-hexadecanoic acid and [2H4]-putrescine were purchased from Campro (Veenendaal, The Netherlands) and [2H6]-salicylic acid was obtained from Icon (Summit, NJ). Stock solutions of each internal standard were prepared in either Milli-Q water or methanol to a concentration of 0.5 μg/μL.
8
Extraction and Characterization of Crocin and Crocetin
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Crocin (PubChem CID: 5281233, purity: 99.33%) and crocetin (PubChem CID: 5281232, purity: 96.61%), Figure 1 , were purchased from Chengdu Biopurify Phytochemicals Ltd. (Chengdu, China). The Compound Edaravone Injection was provided by Jiangsu Simcere Pharmaceutical Co., Ltd. (Nanjing, China). 2,3,5-Triphenyltetrazolium chloride (TTC), stable-isotope-labeled IS 1,2-13C2-myristic acid, ES methyl stearate, methoxyamine hydrochloride, pyridine, hemin, resazurin sodium salt, L -cysteine, 2-ethylbutyric acid, and cholic acid-2,2,4,4-d4 were purchased from Sigma-Aldrich (St. Louis, United States). N-Methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) plus 1% trimethylchlorosilane (TMCS) was purchased from Thermo Scientific (Bellefonte, United States). Streptomycin sulfate and neomycin sulfate were purchased from Nanjing Xinhou Biological Technology Co., Ltd. (Nanjing, China). Yeast extract and tryptone were purchased from Oxoid (Basingstoke, United Kingdom).
9
Lipid Extraction and GC/MS Analysis Protocol
10
Quantitative Analysis of Plasma Isoflavones
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Blood was collected from the abdominal aorta under anesthesia at the time of killing. Plasma was isolated by centrifugation and frozen at -80°C until quantitative analysis of isoflavones (equol, daidzein, and genistein) using gas chromatography mass spectrometry. Sample preparation was performed essentially as described previously, with some modifications. 30 Briefly, 50 μL of a plasma sample was incubated overnight at 37°C with 150 μL of 0.2 mol/L sodium acetate buffer solution (pH 5.5) containing β-glucuronidase/sulfatase (150:1; Sigma-Aldrich) as deconjugating enzymes. Afterward, the isoflavone fraction was extracted with 700 μL of ethyl acetate (Kishida Chemical Co., Ltd., Osaka, Japan) and evaporated to dryness with nitrogen gas, followed by derivatization using 50 μL of N-methyl-N[tert-butyldimethylsilyl trifluoroacetamide] (MTBSTFA) with 1% trimethylchlorosilane (TMCS) (Thermo Fisher Scientific Inc, Waltham, MA) at 100°C for 1.5 hours. After evaporation with nitrogen, the samples were dissolved in hexane and analyzed using gas chromatography mass spectrometry (QP2010, Shimadzu Co, Kyoto, Japan). The detection ranges were 33-6600 nmol/L, 31-6294 nmol/L, and 31-6157 nmol/L for equol, daidzein, and genistein, respectively.
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