BSTFAN,o-Bis (Trimethylsilyl) trifluoroacetamide (derivatization reagent)
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The methoximation reaction was carried out for 90 min while rocking in an air-shaker at 30 [degrees]C, followed by trimethylsilyl for 60 min by adding 80 [micro]L BSTFA at 70 [degrees]C.
Considering that the conventional reagent BSTFA was used for derivatization, the classes of n-alkanols and n-fatty acids were identified by extracting chromatograms for the m/z 103.05729 (exact mass: 103.05737, error: -0.77 ppm) and m/z 117.03663 (exact mass: 117.03653, error: -0.85 ppm), respectively.
Subsequently, the mixture was trimethylsilylated at 37[degrees] C for 1.5 h with 60 [micro]L BSTFA with 1% TMCS.
For derivatization, 100 [micro]L of N,0-Bis-(trimethylsilyl)trifluoroacetamide(BSTFA) (99%) mixture containing trimethylsilylchloride (TMCS) (1%) and 20 [micro]L of pyfidine is added to the silylation vial and the reaction is carried out at 70[degrees]C for 1 h.
Hence, N-methyl-N-(tert-butyldimethylsilyl)trifluoroacetamide (MTB STFA) [24], N,O-bis(trimethylsiliyl)trifluoroacetamide (BSTFA) [27], BSTFA + trimethylchlorosilane (TMCS) (10:1) [29], BSTFA+TMCS (99:1) [13, 21, 30] have been used as silylating reagents while trimethyloxonium tetrafluoroborate has been used as a methylating agent [25, 26].
Trichloroacetic acid (TCA) was purchased from Fisher Scientific, U.K.Trypsin-EDTA solution, Trizma base, sulforhodamine B (SRB) sodium salt, glacial acetic acid, paclitaxel, 3,4-dihydroxybenzoic acid, 4-hydroxy-3-methoxybenzoic acid, gallic acid, trans-catechin, trans-gallocatechin, N,O-bis(trimethylsily) trifluoroacetamide (BSTFA) with 1% chlorotrimethylsilane (TMCS), and pyridine were purchased from Sigma-Aldrich.
Trimethylolpropane (TMP) and N,O-Bis(trimetyhlsilyl)tri-fluroacitemide (BSTFA), Methanol, Hydrocloric acid, Ethyl acetate, orto-Phosphoric acid, and n-hexane were ordered from Sigma Aldrich and Chemmart Asia, Malaysia.
The third fraction was concentrated and evaporated to dryness under a stream of nitrogen gas, treated with bis (trimethylsilyl) trifluoroacetamide (BSTFA) (Sigma Aldrich, Saint Louis, MO) heated at 70[degrees]C for 30 min, and finally re-dissolved with dichloromethane.
A 30 m HP-1 column was used for the analysis of 2,4-DBP, 2,4,6-TBP, and PBP following their silylation with bis-(trimethylsilyl)-trifluoroacetamide (BSTFA) [69].
The trimethylsilyl (TMS) derivatives were performed with BSTFA in acetonitrile instead of THF in the same concentration (1:1 V/V).
Plasma samples drawn during the tracing trial were processed by methoxyamin HCL, BSTFA, TMCS, and pyridine.