Mechanism of retention loss when C8 and C18 HPLC columns are used with highly aqueous mobile phases

Walter, Thomas H.; Iraneta, Pamela C.; Capparella, Mark

doi:10.1016/j.chroma.2005.04.039

Cited by 140 publications

(69 citation statements)

References 22 publications

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“…[25][26][27] A similar retention loss of uric acid and creatinine was also observed on a C18 column with previously reported aqueous mobile phases after two runs, or less than 20 min. The retention losses are attributed to (1) a folding of the stationary phase alkyl chains in the presence of a highly aqueous mobile phase, and (2) the highly aqueous mobile phase being forced out of the pores when the flow is stopped and the pressure released.…”

Section: Chromatographic Separationsupporting

confidence: 53%

“…[17][18][19][22][23][24] However, many studies have reported that the use of a highly aqueous mobile phase (>90% water) with C18 and C8 columns causes an anomalous chromatographic behavior, and a retention loss of analytes. 19,25 These anomalies have been attributed to (i) aggregation of the bonded hydrocarbon chains in the presence of a highly aqueous mobile phase, rendering them inaccessible to analytes, and (ii) extrusion of the highly aqueous mobile phase from the pores of the particles of the stationary phase. [25][26][27] Our preliminary experiments in this study showed a rapid loss in the retention time of creatinine and uric acid on the C18 column with highly aqueous mobile phases reported in some recent publications.…”

Section: Introductionmentioning

confidence: 99%

“…19,25 These anomalies have been attributed to (i) aggregation of the bonded hydrocarbon chains in the presence of a highly aqueous mobile phase, rendering them inaccessible to analytes, and (ii) extrusion of the highly aqueous mobile phase from the pores of the particles of the stationary phase. [25][26][27] Our preliminary experiments in this study showed a rapid loss in the retention time of creatinine and uric acid on the C18 column with highly aqueous mobile phases reported in some recent publications. 19,21 In this paper, a simple, fast, accurate and environmentally friendly reversed-phase HPLC method is described for the simultaneous determination of creatinine and uric acid in human urine.…”

Section: Introductionmentioning

confidence: 99%

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Simultaneous Determination of Creatinine and Uric Acid in Human Urine by High-Performance Liquid Chromatography

et al. 2008

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An environmentally friendly reversed-phase HPLC method for simultaneous determination of creatinine and uric acid in human urine samples has been developed. Human urine samples were pretreated by dilution, protein precipitation, centrifugation and filtration, followed by HPLC separations using a reversed-phase C18 column with an aqueous mobile phase of phosphate buffer. The retention loss of a C18 column when using the highly aqueous mobile phases was avoided by employing a gradient elution using a small volume (<0.23 mL) of acetonitrile and phosphate buffer at pH 4.75. This developed method provides a simple, rapid separation and sensitive detection for the species of interest in 10 min with UV detection at 205 nm. Quantitation was carried out by relating the peak areas of the identified compounds to that of hypoxanthine as an internal standard. The detection limits for creatinine and uric acid were 0.045 and 0.062 mg mL -1 , respectively. The recoveries of the standards added to urine samples were 87.3 -102.2% for creatinine and 97.3 -108.6% for uric acid, and the relative standard deviation for both analytes was less than 1.0%. This method has been successfully applied to estimating of creatinine and uric acid in human urine.

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Section: Chromatographic Separationsupporting

confidence: 53%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Simultaneous Determination of Creatinine and Uric Acid in Human Urine by High-Performance Liquid Chromatography

et al. 2008

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“…In some cases this effect can also be exploited to fully replace the organic co-solvent. However, purging a column with a pure water mobile phase can lead to an unwanted retention loss [56]. In order to circumvent this problem we added 10% of methanol to the purging eluent.…”

Section: General Assessment Of the Resultsmentioning

confidence: 99%

Temperature and pH‐stability of commercial stationary phases

Teutenberg¹,

Hollebekkers²,

Wiese

et al. 2009

J of Separation Science

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Original PaperTemperature and pH-stability of commercial stationary phasesIn this paper, the temperature and pH stability of silica-based RP stationary phases were investigated. Furthermore, nonsiliceous phases like a polymeric column based on polystyrene divinylbenzene and a polybutadiene coated zirconium dioxide column were also included. The columns were heated up to 1508C at dynamic conditions, which means that the eluent consisting of water and methanol (90:10, v/v) was continuously purged through the packed bed. After every 5 h, the columns were cooled down to room temperature and the efficiency was measured by injecting a test sample based on the Neue test. It could be shown that some stationary phases exhibited a very good temperature stability at the test conditions specified above.

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“…They achieve their ability to separate mixtures by retarding the passage of some components while permitting others to move freely, this property can be calculated using retention factor (k) (57,58). Retention factor is a parameter that expresses the ability of a particular solute to interact with a chromatographic system, it relates to the contact surface area of the substance Where B is constant for strictly homologous series, N, Avogadro number and γ is surface tension of solution, R is the gas constant and T the temperature (57).…”

Section: Chromatographymentioning

confidence: 99%

Impact of Surface Tension in Pharmaceutical Sciences

2009

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-Surface chemistry has a large influence in many industries. In the life sciences, surface area is gaining importance in the characterization of materials during their development, formulation and manufacturing. The chemical activity, adsorption, dissolution, and bioavailability of a drug may depend on the surface of the molecule. In order to meet manufacturing challenges and develop new and better performing products with improved qualities, knowledge of surface tension is of utmost importance. An attempt has been made in this paper to review the application of interfacial tension in the key domains of pharmaceutical applications.

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Mechanism of retention loss when C8 and C18 HPLC columns are used with highly aqueous mobile phases

Cited by 140 publications

References 22 publications

Simultaneous Determination of Creatinine and Uric Acid in Human Urine by High-Performance Liquid Chromatography

Simultaneous Determination of Creatinine and Uric Acid in Human Urine by High-Performance Liquid Chromatography

Temperature and pH‐stability of commercial stationary phases

Impact of Surface Tension in Pharmaceutical Sciences

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