scholarly journals Observations on Ion Interaction Chromatographic System: Reversed Phase Column—H3BO3/TBAOH Mobile Phase and the Effect of Temperature

2020 ◽  
Vol 83 (12) ◽  
pp. 1553-1560
Author(s):  
Rajmund S. Dybczyński ◽  
Krzysztof Kulisa
Keyword(s):  
Ion Exchange ◽  
Boric Acid ◽  
Mobile Phase ◽  
Reversed Phase ◽  
Effect Of Temperature ◽  
Chromatographic System ◽  
Retention Loss ◽  
Hydrophobic Adsorption ◽  
Phase Column ◽  
Reversed Phase Column

Abstract New ion interaction chromatographic (IIC) system with RP column and boric acid plus tetra-n-butylammonium hydroxide (TBAOH) mobile phase was investigated. In the system: CPhenylHexyl—10 mM H3BO3/4 mM TBAOH, a large group of anions viz. F−,IO3−, Cl−, ClO2−, BrO3−, NO2−, Br−, NO3−, ClO3−, I−, HPO42−, SO42−, CrO42−, S2O32−, benzoate−, SCN−, ClO4− and phthalate2−, could be isocratically resolved. A study on the effect of temperature revealed that some ions added to ion exchange are also held in the stationary phase by the hydrophobic adsorption. The retention loss for all anions with time was observed. This effect however, was quite slow and good separations could be obtained even after the column stayed a few hundred hours in the mobile phase.

4-Nitrophenol in 4-nitrophenyl phosphate, a substrate for alkaline phosphatase, as measured by paired-ion high-performance liquid chromatography.

1977 ◽  
Vol 23 (12) ◽  
pp. 2288-2291 ◽  
Author(s):  
P H Culbreth ◽  
I W Duncan ◽  
C A Burtis
Keyword(s):  
Alkaline Phosphatase ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Flow Rate ◽  
Mobile Phase ◽  
High Performance ◽  
Reversed Phase ◽  
Nitrophenyl Phosphate ◽  
Phase Column ◽  
Reversed Phase Column

Abstract We used paired-ion high-performance liquid chromatography to determine the 4-nitrophenol content of 4-nitrophenyl phosphate, a substrate for alkaline phosphatase analysis. This was done on a reversed-phase column with a mobile phase of methanol/water, 45/55 by vol, containing 3 ml of tetrabutylammonium phosphate reagent per 200 ml of solvent. At a flow rate of 1 ml/min, 4-nitrophenol was eluted at 9 min and monitored at 404 nm; 4-nitrophenyl phosphate was eluted at 5 min and could be monitored at 311 nm. Samples of 4-nitrophenyl phosphate obtained from several sources contained 0.3 to 7.8 mole of 4-nitrophenol per mole of 4-nitrophenyl phosphate.

scholarly journals HPLC FINGERPRINT OF ORGANIC ACIDS IN FRUIT JUICES

1970 ◽  
Vol 62 ◽  
Author(s):  
Loredana Leopold ◽  
Diehl Horst ◽  
Carmen Socaciu
Keyword(s):  
Organic Acids ◽  
Stationary Phase ◽  
Organic Acid ◽  
Mobile Phase ◽  
Reversed Phase ◽  
Fruit Juices ◽  
Hplc Fingerprint ◽  
Organic Acid Profile ◽  
Phase Column ◽  
Reversed Phase Column

Organic acids give fruit products their characteristic tartness and vary in combination and in concentrations among different juices. The organic acid profile can be used to identify a juice or verify its purity. Typically, organic acids in fruit juices are identified and quantified by using methods such as HPLC. In this procedure, reversed phase column is used to separate and identificate six organic acids. Because several of the analytes are extremely difficult to resolve, a aqueous mobile phase is needed to enhance interaction between the acids and the C18 stationary phase.

Liquid-chromatographic quantification of plasma phenylalanine, tyrosine, and tryptophan.

1981 ◽  
Vol 27 (1) ◽  
pp. 146-148 ◽  
Author(s):  
L M Neckers ◽  
L E Delisi ◽  
R J Wyatt
Keyword(s):  
Amino Acids ◽  
High Pressure ◽  
Liquid Chromatography ◽  
Fluorescence Detection ◽  
Mobile Phase ◽  
Reversed Phase ◽  
Plasma Phenylalanine ◽  
Liquid Chromatographic ◽  
Phase Column ◽  
Reversed Phase Column

Abstract Phenylalanine, tyrosine, and tryptophan are isolated and quantified by "high-pressure" liquid chromatography, with fluorescence detection. An isocratic mobile phase and reversed-phase column are used to provide rapid and reproducible measurement of these amino acids in as little as 1 to 2 microL of human plasma.

Determination of bethanidine in plasma by liquid-chromatography with a microbore reversed-phase column.

1983 ◽  
Vol 29 (10) ◽  
pp. 1793-1795 ◽  
Author(s):  
J R Shipe ◽  
A F Arlinghaus ◽  
J Savory ◽  
M R Wills ◽  
J P DiMarco
Keyword(s):  
Liquid Chromatography ◽  
Mobile Phase ◽  
Internal Standard ◽  
Reversed Phase ◽  
Ultraviolet Absorbance ◽  
Standard Curve ◽  
Novel Method ◽  
Phase Column ◽  
Reversed Phase Column

Abstract In this novel method for quantifying bethanidine in plasma, after a multi-step extraction of bethanidine and internal standard from 2.0 mL of plasma, the drugs are separated on a "microbore" C18 reversed-phase column and quantified by their ultraviolet absorbance at 210 nm. The isocratic chromatographic separation takes about 15 min with use of an ion-pairing regent in the mobile phase (acetate buffer/acetonitrile, 9/1 by vol) and a flow rate of 0.25 mL/min. Sensitivity is increased relative to conventional columns, and solvent consumption is reduced by 90%. The standard curve is linear to at least 5 mg/L, and the detection limit is 0.02 mg/L. The within-run precision of the method is excellent (CV 4%) at a midrange concentration of 1.25 mg/L.

Liquid-chromatographic measurement of phenylalanine and tyrosine in serum.

1982 ◽  
Vol 28 (11) ◽  
pp. 2282-2285 ◽  
Author(s):  
F W Spierto ◽  
W Whitfield ◽  
M Apetz ◽  
W H Hannon
Keyword(s):  
Mobile Phase ◽  
High Performance ◽  
Phenylalanine Ammonia Lyase ◽  
Reversed Phase ◽  
Coumaric Acid ◽  
Chromatographic Measurement ◽  
Liquid Chromatographic ◽  
Detection And Quantification ◽  
Phase Column ◽  
Reversed Phase Column

Abstract With phenylalanine ammonia-lyase (EC 4.3.1.5) we converted phenylalanine (Phe) and tyrosine (Tyr) to transcinnamic acid and p-coumaric acid, respectively. These were separated by "high-performance" liquid chromatography and detected at 280 nm. We measured the Phe and Tyr content of human serum by adding 100 mU of the enzyme to a 20-microL serum aliquot, mixing for 2 h at 24 degrees C, then stopping the reaction with 1 mL of cold methanol. Precipitated proteins were removed by centrifugation and the separated clear supernates were stored at -20 degrees C. For chromatographic separation, detection, and quantification, we used a system equipped with a C-18 reversed-phase column, a variable-wavelength spectrophotometer, a printer-plotter, and a microcomputer. The mobile phase was a mixture of dilute aqueous (50 g/L) acetic acid and CH3CN (80/20, by vol). CVs for specimens containing 100 mg of Phe or Tyr per liter varied from 5 to 10%. Analytical recoveries were near 100%.

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