Triethylammonium bicarbonate

Adenosine 5"-[P-thio]diphosphate tri-lithium salt [73536-95-5] M 461.1. Purified by ion-exchange chromatography on DEAE-Sephadex A-25 using gradient elution with 0.1-0.5M triethylammonium bicarbonate. [Biochem Biophys Acta 276 155 7972.]... 

Workup (a) The product is isolated by passage through an ion exchange column (Dowcx AG1-X8, HCOf 100 mL) and consecutive elution with 200 mM triethylammonium bicarbonate solution. After ion exchange... 

Tubes for collecting fractions 100 mM triethylammonium acetate, pH 7.0, in water 10 mM triethylammonium bicarbonate, pH 7.0, in water... 

Nucleotide thiophosphate analogues. The preparation and purification of [ HJATPyS, pHJGTPyS, s ITPyS (6-thioinosine), cl ITPyS (6-chloroinosine) and [ HJATPyS are described and the general purification was achieved by chromatography of the nucleotide thiophosphates in the minimum volume of H2O placed onto a DEAE-Sephadex A25 column and eluting with a linear gradient of triethylammonium bicarbonate (0.1 to 0.6M for G and I nucleotides and 0.2 to 0.5M for A nucleotides). [Biochim Biophys Acta 276 155 7972]. 

Volatile buffers were reconsidered for the modified method. Triethylamine was ruled out primarily because it could not be obtained in high purity and because the secondary and primary amines contaminating it could potentially react with solutes present in the water sample. Preliminary evidence of reaction between ethidium bromide and triethylammonium bicarbonate was obtained, but the reaction product was not characterized. The components of volatile buffers that appeared acceptable on the basis of chemical purity were ammonia, acetic acid, and formic acid. A few exploratory experiments were conducted involving the elution by ammonium formate and ammonium acetate of EB or quinaldic acid exchanged onto AG MP-50 or IRA 900. These experiments showed that 1 M ammonium formate in water was a very poor eluent, but that EB could be eluted from AG MP-50 with 1 M ammonium formate in methanol. Elution was essentially complete with 6 bed volumes of the methanolic eluent, whereas neither methanol alone nor aqueous 1 M ammonium formate was able to elute this solute. This situation pointed out the necessity for a counterion to displace exchanged solutes and, additionally, indicated that the displaced solute be highly soluble in the eluting solvent. 

A monolithic silica-based CIS stationary phase was used under high flow rate condition (2 mL/min) without significant back pressure in IPC analysis of a recently discovered new drug candidate for the treatment of Alzheimer s disease [15]. Nanoscale IPC using a monolithic poly(styrene-divinylbenzene) (PS-DVB) nanocolumn coupled to nanoelectrospray ionization mass spectrometry (nano-ESl-MS) was evaluated to separate and identify isomeric oligonucleotide adducts. Triethylammonium bicarbonate was used as the IPR. Interestingly, the performance of the polymeric monolithic PS-DVB stationary phase significantly surpassed that of columns packed with the microparticulate sorbents CIS or PS-DVB [16]. 

The residue then was dissolved in 30 ml of 0.04 M aqueous triethyl-ammonium bicarbonate buffer (pH 7.5), (resulting pH 5.6). It was chromatographed on a DEAE cellulose column (HCOJ form) (3.4 X 15.0 cm), with elution first with 500 ml of water and then with a linear gradient of trimethylammonium bicarbonate at pH 7.2 (0-0.07 M). XXVII was completely resolved from some of XXVIII which was formed under the above reaction conditions. However, it was still contaminated by small amounts of cAMP and ethyl 2-diazo-malonic acid. These impurities were apparently generated from the hydrolysis of some XXVII when the triethylammonium bicarbonate was removed. Pure samples of XXVII were obtained by preparative paper chromatography on either Whatman 40 or 3 MM paper with overnight development with ethanol 0.5 M ammonium acetate, pH 7.0 (5 2, v/v). 

A TLC- and HPLC-based method was developed to quantitatively detect the cytokinins and auxins produced in the acid and alkaline extracts of culture filtrates from different P. amygdali strains. The IAA and cytokinin (t-Z and t-ZR) quantification was performed by an HPLC method developed using a C-18 reverse phase column and a linear gradient of MeCN-H20 (7 3, v/v) for the analysis of IAA, and 100 pM TEAB (triethylammonium bicarbonate adjusted at pH=7 with COf) and MeCN-H20 (7 3, v/v) in steps 1 and 2 for the analysis of cytokinins. The presence and level of phytohormones produced by some strains were analysed with respect to their virulence on almond plants [41]. The same analytical methods were used to estimate the production of cytokinins and/or auxins by three wild strains of P. savastanoi and three phytohormone-deficient mutants. The pathogeneticity on olive and oleander plants of three wild-type strains of P. savastanoi (two from olive and one from oleander) was compared to those of three phytohormone-deficient mutants of oleander strains Iaa+/cytokinin, Iaa-... 

Fig. 3. Reverse phase HPLC of a mixture of naturally-occurring cytokinins. Column 150x4.6 mm 5 p.m Spherisorb ODS-2. Mobile phase 30 min, 5-20% acetonitrile in water (pH 7.0 with triethylammonium bicarbonate). Flow rate 2 ml min. Detector absorbance monitor at 265 nm. Sample (1) adenine, (2) adenosine, (3) zeatin-7-glucoside, (4) zeatin-9-glucoside. (5) zeatin-O-glucoside, (6) zeatin, (7) dihydrozeatin-O-glucoside, (8) dihydrozeatin, (9) dihydrozeatin riboside-O-glucoside, (10) zeatin riboside, (11) dihydrozeatin riboside [6].

Alkylation reaction 45 pi of acetonitrile, 15 pi of triethylammonium bicarbonate solution (pH 8,5) and 14 pi of methyhodide were added. The reaction was incubated at 40°C for 25 min. Afterwards 20 pi double distilled water were added. Upon cooling a biphasic system was obtained. The upper layer contained the products while the lower layer contained some of the reagents, for example detergents that were added to stabilise the enzymes used in this procedure. 20 pi of the upper layer were sampled and diluted in 45 pi of 40% acetonitrile. This solution was directly used to transfer the samples onto the matrix. 

Deprotection of the 5 -phosphate is easily accomplished after oxidative treatment. Oxidation with sodium metaperiodate in water (or in 0.1 M triethylammonium bicarbonate) gives the corresponding sulfoxide derivative. The free nucleotide is hberated by treatment with 2 M aqueous sodium hydroxide at room temperature (eq 2). Alternatively, the sulfide is oxidized to suhbne with NCS in 0.1 M triethylammonium bicarbonate buffer at room temperature. In this case, deprotection can be achieved successfully... 

To remove the yellow color, the material (approximately 2 mmoles) may be applied to a Dowex 1X4 ion-exchange column (1.7 X 17 cm, OH form) and the column is washed sequentially with H2O (500 ml) and with 50% aqueous MeOH (500 ml) the compound is eluted with 0.1 Af triethylammonium bicarbonate. The eluate is evaporated, and the buffer is removed by repeated evaporations with MeOH. The residue is triturated with acetone (20 ml) and filtered the acetone solution is evaporated, the residue is applied to a SiOj column (1.7 X 17 cm), and the column is eluted with acetone. The acetone solution is evaporated the residue crystallizes on standing white needles, m.p. 139°-147 , with darkening and decomposition which became very rapid above 180°. Yield 95%. 

The tubes containing [ P]8-Br-cAMP are pooled and evaporated to dryness at 30° under reduced pressure with a Buchi rotovaporator. The material is dissolved and coevaporated four times with methanol to remove excess triethylammonium bicarbonate. An ultraviolet spectrum of the product has a Amax of 264 as expected for 8-Br-cAMP. The [ P]8-Br-cAMP is placed in a 25-ml round-bottomed flask with 25 ml of dimethylformamide containing 10 mmoles of triethylammonium azide. The last solution is prepared by passing 500 mg of LiNa (10 mmoles), dissolved in dimethylformamide, through a 20-ml Dowex 50 column in the triethylammonium form and also in dimethylformamide as solvent. 

Of the anomeric mixture of fo" UMP, 10,500 A-,r,u units are converted to their triphosphates by a standard procedure.. The crude reaction product is charged onto a column of DEAE-Sephadex and eluted with a linear gradient of 2.5 liters each of 0.05 M and 0.45 M triethylammonium bicarbonate. The a-fo UTP (856 A,so units) elutes between 0.39 and 0.42 M buffer and the j3-anomer (1190 units) between 0.35 and... 

Mehta et al. (26) separated guanine on PEI-cellulose with triethylammonium bicarbonate (TEAB) 0.5 M pH 7.6. Good separation of cyclics, phosphates and nucleosides was evident. Trifilo and Dobson (27) separated cyclic purines by PEI, ammonium acetate/hydroxide-ethanol at pH 9.0. One dimension, ascending, from triphosphates to nucleobases. Gulyassy and Farrand (28) also separated cyclic purines with PEI in 0.4 M acetic, then 0.125 M LiCl. Manhandhar and Dyke (29) separated GTP with PEI and luciferase, water and then 1.4 M LiCl for 50 min. Assay was by scintillation counter. 

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