Charged Aerosol Detector Research Articles

Simple and Direct Analysis of Falcarinol and other polyacetylenic Oxylipins in Carrots by Reverse Phase HPLC and Charged Aerosol Detection

Simple and Direct Analysis of Falcarinol and other polyacetylenic Oxylipins in Carrots by Reverse Phase HPLC and Charged Aerosol Detection

Sensitive Analysis of Commonly Used Artificial and Natural Sweeteners Including Stevia and Their Impurities and Degradation Products

Many recently commercialized sweeteners tend to have increased potency, reducing the amount of active ingredient added to beverages and other food products and often providing cost savings to the manufacturer. Using increased potency has also contributed to a need for sensitive analytical methods to quantify the active product and to detect low levels of breakdown products and impurities, which are required for quality and safety issues. Because compounds typically do not possess any chromophore, traditional HPLC-UV approaches are inappropriate. The work in this study describes several methods using HPLC with the Thermo Scientific Dionex CoronaTM Charged Aerosol Detector (CADTM) for the study of common natural sugars (fructose, glucose, turanose, saccharose, trehalose, maltose, melezitose, and raffinose); artificial sweeteners (sucralose, aspartame, saccharin, and acesulfame K); and newly introduced products containing stevia extracts (rebaudioside A and stevioside). These HPLC-CAD methods provide sensitivity at low levels (ng) with good reproducibility and accuracy, and correlation to the component concentrations. Stevia products were analyzed by both charged aerosol detection and UV. Charged aerosol detection showed a greater than fivefold improvement in sensitivity over UV for all major components. Finally, the UHPLC methods developed showed a decreased run time and an increased sensitivity for glucose, lactose, and sucrose. Typical limits of detection (LOD) were found to be <500 pg (on column) for glucose and other monoand disaccharides. HPLC-CAD is a very flexible approach to measuring sweeteners and overcomes many of the limitations of UV, refractive index (RI), LCMS, evaporative light scattering detection (ELSD), and HPLC-pulsed amperometric approaches. The HPLC-CAD platform can be used throughout the manufacturing process to ensure finished quality and batch-to-batch uniformity. Introduction The Dionex Corona CAD is a mass-sensitive universal detector for nonvolatile and some semivolatile analytes. Unlike absorbance, fluorescence, RI, or electrochemical detection, analyte response is independent of chemical Sensitive Analysis of Commonly Used Artificial and Natural Sweeteners Including Stevia and Their Impurities and Degradation Products Ian Acworth, Chris Crafts, and Bruce Bailey, Thermo Fisher Scientific, Chelmsford, MA, USA structure. Carbohydrates lack chromophores and the typical approaches used for their analysis may lack sensitivity, require derivatization, or cannot be used with gradient. The work in this study examines different HPLC-CAD methods for the analysis of honey sugars, artificial sweeteners, and other natural products including stevia extracts. Finally, a new UHPLC method for fast analysis of simple carbohydrates is presented. The Corona CAD system is easy to use and offers a simple, sensitive, reproducible, and direct approach for the routine analysis of natural and artificial sweeteners. Artificial Sweeteners Global Method HPLC Parameters Column: ACE®, C18 4.6 × 250 mm, 5 μm Mobile Phase: A) Deionized water B) Acetonitrile + 0.1% trifluoroacetic acid Gradient: 2 to 40% B over 25 min, 40 to 60% 25–30 min Flow Rate: 1.0 mL/min Inj. Volume: 50 μL Column Temp.: 30 °C Samples: 1.2 to 20 μg on column FIGURE 1. Chromatogram of artificial sweeteners. 26831 5 10 15 20 25 30 35 400 A ce su lfa m e K C yc la m at e S ac ch ar in S uc ra lo se A sp ar ta m e

Simultaneous determination of Maillard reaction impurities in memantine tablets using HPLC with charged aerosol detector

A gradient high performance liquid chromatographic (HPLC) method using charged aerosol detection (CAD) was developed for the simultaneous determination of impurities formed by the Maillard reaction in memantine tablets. These impurities were a memantine-lactose adduct (ML), a memantine-dimethylamino glycine adduct (DMAG), a memantine-galactose adduct (MGAL), and a memantine-glucose adduct (MGLU). The chromatographic separation was performed on a Synergy Hydro RP column (100 mm × 3 mm, 2.5 μm particles) from Phenomenex with gradient elution using mobile phases consisting of 0.6% (v/v) of heptafluorobuturic acid (HFBA) in two acetonitrile–isopropyl alcohol–water mixtures. The method validation for the impurities included linearity, accuracy by spike recovery, precision, limits of detection and quantitation, and robustness. The method was sensitive for these non-chromophoric impurities down to 0.4–0.6 μg/mL (0.02–0.03% of the memantine drug substance). The effect of mobile phase HFBA concentration on chromatographic resolution and peak shape was investigated, and the effect of sample diluent acidification on method accuracy via spike recovery was studied. The operational simplicity of the CAD detector for routine quality control has been demonstrated.

Composition of structural carbohydrates in biomass: Precision of a liquid chromatography method using a neutral detergent extraction and a charged aerosol detector

We adapted and optimized a method to quantify the cellulose, hemicellulose, xylan, arabinan, mannan, galactan contents in lignocellulosic biomass. This method is based on a neutral detergent extraction (NDE) of the interfering biomass components, followed by a sulfuric acid hydrolysis (SAH) of the structural polysaccharides, and a liquid chromatography with charged aerosol detection (LC-CAD) to analyze the released monosaccharides. The first step of this NDE-SAH-LC-CAD method aims at removing all compounds that interfere with the subsequent sulphuric acid hydrolysis or with the subsequent chromatographic quantification of the cellulosic and hemicellulosic monosaccharides. This step includes starch hydrolysis with an analytical thermostable α-amylase followed by an extraction of soluble compounds by a Van Soest neutral detergent solution (NDE). The aim of this paper was to assess the precision of this method when choosing fiber sorghum (Sorghum bicolor (L.) Moench), tall fescue (Festuca arundinacea Schreb.) and fiber hemp (Cannabis sativa L.) as representative lignocellulosic biomass. The cellulose content of fiber sorghum, tall fescue and fiber hemp determined by the NDE-SAH-LC-CAD method were 28.7±1.0, 29.7±1.0 and 43.6±1.2g/100g dry matter, respectively, and their hemicellulose content were 18.6±0.5, 16.5±0.5 and 14.5±0.2g/100g dry matter, respectively. Cellulose, mannan and galactan contents were higher in fiber hemp (dicotyledon) as compared to tall fescue and fiber sorghum (monocotyledons). The xylan, arabinan and total hemicellulose contents were higher in tall fescue and fiber sorghum as compared to fiber hemp. The precision of the NDE-SAH-LC-CAD method was better for polysaccharide concentration levels above 1g/100g dry matter. Galactan analysis offered a lower precision, due to a lower CAD response intensity to galactose as compared to the other monosaccharides. The dispersions of the results (expanded uncertainty) of the NDE-SAH-LC-CAD method were smaller as compared to the Van Soest (VS) method. In addition, the NDE-SAH-LC-CAD method was able to provide additional information on the composition of the hemicellulose (xylan, arabinan, mannan and galactan content) that is not provided by the Van Soest method. The NDE-SAH-LC-CAD method offers also the advantage of a better specificity for hemicellulose and cellulose, as compared to the NREL and Uppsala methods.

Isolation of Pure Phospholipid Fraction from Egg Yolk

AbstractThe phospholipid (PL) fraction from egg yolk was isolated and purified. In the procedure applied (method 2) the egg yolk was extracted with ethanol, precipitated using acetone chilled to −20 °C and washed using acetone. The purity of the samples was checked by HPLC analysis using a Charged Aerosol Detector (CAD). The results were compared with those obtained for the phospholipid fraction isolated and purified by deoiling yolk before extraction and the precipitation of PL with acetone chilled to 4 °C (method 1). The use of acetone chilled to −20 °C to precipitate and wash the phospholipids yielded the phospholipid fraction with 100% purity (78.7 ± 0.2 of phosphatidylcholine and 21.3 ± 0.2 of phosphatidylethanolamine). When deoiling and the 4 °C purification process was used (method 1) 0.4 ± 0.1% cholesterol and some traces of triacylglycerols remained in the PL fraction.

Development of a Reversed-Phase HPLC Impurity Method for a UV Variable Isomeric Mixture of a CRF Drug Substance Intermediate with the Assistance of Corona CAD

The compound studied, an antagonist of corticotropin-releasing factor (CRF) receptor 1, was under development for the treatment of major depressive disorder and generalized anxiety disorder. To control the quality of intermediate compounds in the synthetic pathway, the preference is to develop a single HPLC method that could determine impurities of the sequential and structurally similar intermediates. However, the chemical nature of one of those intermediates, Compound C, posed a number of challenges to the development of a suitably rugged impurity method for this isolated intermediate. In the solid state, Compound C is a stable mixture of its trans-isomer and semicarbazone forms. In most solution states, Compound C interchanges between the trans and semicarbazone forms as well as degrades rapidly to impurity X, resulting in sample solution instability. Furthermore, the trans-isomers and semicarbazone exhibited different UV response factors, but reference standards of the two forms were not available to determine the relative response factor. Consequently, irreproducibility was observed during the HPLC analysis when conventional UV detection was applied directly. This article describes the following approaches that were employed to meet these challenges: (1) diluent screening was performed to select a \solvent that afforded adequate sample solution stability; (2) appropriate chromatographic conditions were selected to eliminate or minimize any on-column degradation; (3) alternate HPLC detection, Corona charged aerosol detection, in conjunction with UV analysis, was used to determine relative amounts of the isomeric forms which subsequently permitted the use of conventional UV detection for routine analysis. The validation data of this reversed-phase liquid chromatographic method are also discussed.

Identification and control of impurities in streptomycin sulfate by high-performance liquid chromatography coupled with mass detection and corona charged-aerosol detection

For the control of impurities in streptomycin sulfate a reversed phase ion-pair high performance liquid chromatography (HPLC) method using charged aerosol detection (CAD) was developed. With this method, 21 impurities could be separated and tentatively identified using a combination of exact mass measurement by TOF-MS and MS/MS experiments with a triple quadrupole MS. For three impurities the suggested structures could be confirmed by in situ formation. The CAD detector response was found to be linear over 2 orders of magnitude allowing a straightforward quantification of all impurities. A limit of quantification of 0.09% for streptomycin sulfate and of 0.008% for streptidine sulfate (referred to the concentration of the 5 mg/ml test solution) could be achieved. The HPLC method was applied to the purity testing of 12 samples of commercially available streptomycin sulfate from different manufacturers. Impurity levels between 4.6% and 16.0% were found. The current European Pharmacopoeia monograph for streptomycin sulfate only limits streptomycin B by a TLC test to 3.0%. Therefore, the results of this study underline the importance of introducing a state-of-the-art test for the control of impurities in the monograph. The new HPLC-CAD method is considered suitable for this purpose.

Characterization of hydroxypropylmethylcellulose (HPMC) using comprehensive two-dimensional liquid chromatography

Various hydroxyl-propylmethylcellulose (HPMC) polymers were characterized according to size and compositional distributions (percentage of methoxyl and hydroxyl-propoxyl substitution) by means of comprehensive two-dimensional liquid chromatography (LC × LC) using reversed-phase (RP) liquid chromatography in the first dimension and aqueous size-exclusion chromatography (aq-SEC) in the second dimension. RP separation was carried out in gradient-elution mode applying 0.05% TFA in water and 1-propanol, while 0.05% TFA in water was used as mobile phase in aqueous SEC. A two-position ten-port switching valve equipped with two storage loops was used to realize LC × LC. Detection of HPMC was accomplished by charged-aerosol detection (CAD). Data processing to visualize chromatograms was carried out using Matlab software. The significant influence of the LC × LC temperature on (the retention of) HPMC was studied using a column oven which allowed accurate temperature control. Due to the phenomenon of thermal gelation, which is a result of methyl and hydroxypropyl substitution of anhydroglucose units from the cellulose backbone, we were able to obtain additional, specific information on compositional characteristics of various HPMC samples. As the retention behaviour of gelated and non-gelated polymer proved to be different, the fraction of the polymer that is gelated in the chromatographic column could be monitored at different temperatures. Moreover, the temperature at which half of the polymer is gelated could be correlated with the cloud-point temperature. As a result, differences in inherent cloud points of modified cellulose can be used as a further distinguishing property in “temperature-responsive” LC × LC.

Investigating the Stability of the Nonionic Surfactants Tocopheryl Polyethylene glycol Succinate and Sucrose Laurate by HPLC–MS, DAD, and CAD

High-performance liquid chromatography (HPLC) methods using a charged aerosol detector (CAD), a mass selective detector (MSD), and a diode array detector (DAD) were developed to characterize the nonionic surfactants d-α-tocopheryl polyethylene glycol (1000) succinate (TPGS) and Surfhope sugar ester D-1216 (sucrose laurate). The molecular structure and the heterogeneous composition resulting from different isomers and various lengths of polyethylene glycol (PEG) chains make it difficult to develop sensitive and specific analytical methods for both surfactants. Hence, there is lack of knowledge about the stability and grade of impurity of these compounds. Sucrose laurate does not possess any chromophore, thus UV detection is not applicable. Therefore, CAD and MSD have been used for determination. The aim of the study was to characterize these nonionic surfactants and to examine chemical stability at pH 1.0 and 37 °C, simulating harsh gastric conditions. It was shown that both compounds are liable to degradation under these conditions. Sucrose monolaurate exhibited a massive degradation within 8 h incubation due to cleavage of the glycosidic bondage. About 50% of sucrose monolaurate broke down, whereas a marginal amount of 3.4% (± 0.4%) of TPGS degraded into d-α-tocopheryl succinate and the associated PEG chain.

Comparison of two aerosol-based detectors for the analysis of gabapentin in pharmaceutical formulations by hydrophilic interaction chromatography

Comparison of hydrophilic interaction chromatography (HILIC) columns coupled with an evaporative light scattering detector (ELSD) or charged aerosol detector (CAD) was done for the detection of gabapentin in pharmaceutical formulations. The chromatographic separations were achieved on four HILIC columns: ZIC HILIC, ZIC pHILIC, Luna HILIC, and Atlantis HILIC. Experimental factors such as mobile phase composition, acetonitrile content, and mobile phase pH were evaluated. Validation of method was done in terms of linearity, sensitivity, accuracy, and precision. The performance of ELSD detection method is comparable to that of CAD. The intra-day and inter-day variations were below 1.7% and 3.2% for CAD and 2.8%, and 3.4% for ELSD, respectively. In addition, detection sensitivities of ELSD, CAD, and UV detectors were also compared for HILIC and reversed phase (RP) modes and the highest sensitivities were obtained in the HILIC mode when connected with CAD and ELSD. The developed HILIC aerosol based detection methods were successfully applied to the analysis of gabapentin in commercial tablets and capsules.

‘Dilute‐and‐Shoot’ Method for Vitamin D Analysis in Dietary Supplement Capsules

Three methods are demonstrated for a complete analysis of vitamin D and triacylglycerols (TAGs) in fortified dietary supplements that virtually eliminates all chemical pretreatment prior to analysis. Three mass spectrometers, in parallel, plus a UV detector, an evaporative light scattering detector (ELSD), and a corona charged aerosol detector (CAD), were used simultaneously. The contents of gelcaps labeled as containing 1000 IU (25 mcg) vitamin D3 in safflower oil and 2000 IU (50 mcg) vitamin D3 in rice bran oil were analyzed without the need for lengthy saponification and extraction. Three to five gelcaps were analyzed using vitamin D2 as an internal standard. Vitamin D3 was analyzed using UV detection, selected ion monitoring (SIM) atmospheric pressure chemical ionization mass spectrometry (APCI‐MS), and two transitions of multiple reaction monitoring (MRM) APCI‐MS. TAGs in the oils were analyzed using full‐scan APCI‐MS, electrospray ionization (ESI) MS, up to MS4, plus two auxiliary detectors‐‐‐the ELSD and CAD. TAGs containing fatty acids up to 28 carbons in length were identified. The vitamin D contents of the gelcaps exceeded label values, and additional chromatographic peaks appeared when the source of vitamin D3 was fish oil (safflower oil gelcap) versus synthetic vitamin D3 (rice bran oil gelcap).

Olive oil quantification of edible vegetable oil blends using triacylglycerols chromatographic fingerprints and chemometric tools

The present work studies the effectiveness of the use of triacylglycerols (TAGs) for the quantification of olive oil in blends with vegetable oils. The determinations were obtained using high-performance liquid chromatography (HPLC) coupled to a Charged Aerosol Detector (CAD), in combination with Partial Least Squares (PLS) regression and using interval PLS (iPLS) for variable selection.Results revealed that PLS models can predict olive oil concentrations with reasonable errors. Variable selection through iPLS did not improve predictions significantly, but revealed the chemical information important in the chromatogram to quantify olive oil in vegetable oil blends.

Quantification of Triacylglycerols in Olive Oils Using HPLC-CAD

This paper describes a reversed phase high-performance liquid chromatography method for the quantification of triacylglycerol (TAG) compositional profile in olive oils. The chromatographic separation was achieved using a C18 column, and the detection was accomplished using a charged aerosol detector. TAG peaks were identified using retention times and by addition of TAG standards. The linearity, matrix effect, and accuracy (precision and trueness) were validated; detection and quantification limits of the method were estimated. The developed analytical method was applied to the determination of the TAG composition and the estimation of the concentration of each individual TAG, using an internal quantification standard (tristearin). From the TAG compositional profile, a fatty acid compositional profile could be calculated. The proposed method appears to be rapid and simple to determine TAGs in olive oil.

Discriminating olive and non-olive oils using HPLC-CAD and chemometrics

This work presents a method for an efficient differentiation of olive oil and several types of vegetable oils using chemometric tools. Triacylglycerides (TAGs) profiles of 126 samples of different categories and varieties of olive oils, and types of edible oils, including corn, sunflower, peanut, soybean, rapeseed, canola, seed, sesame, grape seed, and some mixed oils, have been analyzed. High-performance liquid chromatography coupled to a charged aerosol detector was used to characterize TAGs. The complete chromatograms were evaluated by PCA, PLS-DA, and MCR in combination with suitable preprocessing. The chromatographic data show two clusters; one for olive oil samples and another for the non-olive oils. Commercial oil blends are located between the groups, depending on the concentration of olive oil in the sample. As a result, a good classification among olive oils and non-olive oils and a chemical justification of such classification was achieved.

Recent Advances in the Analysis of Organic Impurities of Active Pharmaceutical Ingredients and Formulations: A Review

Impurity profiling of active pharmaceutical ingredients is a critical quality control parameter during all stages of the development, production and stability testing of pharmaceutical formulations. Literature on this is continuously increasing, it is therefore important to keep track on the latest achievements and applications. This review presents an overview of the most recently published results on the analysis of organic impurities in pharmaceutical samples and active ingredients. Special attention is given to the most widely used techniques, such as liquid chromatography, capillary electrophoresis and mass spectrometry. Keywords: Impurity profiling, Analytical techniques, Stability-indicating assays, Active pharmaceutical ingredients, betamethasone-E-enolaldehyde, betamethasone-Z-enolaldehyde, Charged Aerosol Detector, CAD, mildronate, hydrophilic interaction chromatography, HILIC, Capillary electrophoresis, CE, cyclodextrin derivatives, Normal Stacking Mode, NSM, Reverse Migrating Micelles Mode, SRMM, APCI, Atmospheric Pressure Chemical Ionization, Atmospheric Pressure Ionization, Background Electrole, Charge Aerosol Detector, Micellar Electrokinetic Chromatography, Pseudostationary Phase, Thermal Surface Ionization

Selectivity issues in targeted metabolomics: Separation of phosphorylated carbohydrate isomers by mixed‐mode hydrophilic interaction/weak anion exchange chromatography

Phosphorylated carbohydrates are important intracellular metabolites and thus of prime interest in metabolomics research. Complications in their analysis arise from the existence of structural isomers that do have similar fragmentation patterns in MS/MS and are hard to resolve chromatographically. Herein, we present selective methods for the liquid chromatographic separation of sugar phosphates, such as hexose and pentose phosphates, 2- and 3-phosphoglycerate, dihydroxyacetone phosphate and glyceraldehyde 3-phosphate, as well as glucosamine 1- and 6-phosphate utilizing mixed-mode chromatography with reversed-phase/weak anion-exchangers and a charged aerosol detector. The best results were obtained when the reversed-phase/weak anion-exchanger column was operated under hydrophilic interaction liquid chromatography elution conditions. The effects of various chromatographic parameters were examined and are discussed on the basis of a simple stoichiometric displacement model for explaining ion-exchange processes. Employed acidic conditions have led to the complete separation of α- and β-anomers of glucose 6-phosphate at low temperature. The anomers coeluted in a single peak at elevated temperatures (>40°C) (peak coalescence), while at intermediate temperatures on-column interconversion with a plateau in-between resolved anomer peaks was observed with apparent reaction rate constants between 0.1 and 27.8×10(-4) s(-1). Dynamic HPLC under specified conditions enabled to investigate mutarotation of phosphorylated carbohydrates, their interconversion kinetics, and energy barriers for interconversion. A complex mixture of six hexose phosphate structural isomers could be resolved almost completely.

Determination of Gentamicin Sulphate Composition and Related Substances in Pharmaceutical Preparations by LC with Charged Aerosol Detection.

A new, simple and repeatable liquid chromatography method with charged aerosol detection (LC-CAD) for the determination of gentamicin sulphate composition and related substances has been developed. Gentamicin lacks of chromophores, therefore its determination is quite problematic. Using a universal CAD enables to achieve good separation without sample derivatization. Mass spectrometry was employed to confirm the LC-CAD peak profile. The proposed method was validated and applied for the determination of gentamicin sulphate composition and related substances in pharmaceutical preparations.

Universal response model for a corona charged aerosol detector

The universality of the response of the Corona Charged Aerosol Detector (CoronaCAD) has been investigated under flow-injection and gradient HPLC elution conditions. A three-dimensional model was developed which relates the CoronaCAD response to analyte concentration and the mobile phase composition used. The model was developed using the response of four probe analytes which displayed non-volatile behavior in the CoronaCAD and were soluble over a broad range of mobile phase compositions. The analyte concentrations ranged from 1 μg/mL to 1 mg/mL, and injection volumes corresponded to on-column amounts of 25 ng to 25 μg. Mobile phases used in the model were composed of 0–80% acetonitrile, mixed with complementary proportions of aqueous formic acid (0.1%, pH 2.6). An analyte set of 23 compounds possessing a wide range of physicochemical properties was selected for the purpose of evaluating the model. The predicted response was compared to the actual analyte response displayed by the detector and the efficacy of the model under flow-injection and gradient HPLC elution conditions was determined. The average error of the four analytes used to develop the model was 9.2% ( n = 176), while the errors under flow-injection and gradient HPLC elution conditions for the evaluation set of analytes were found to be 12.5% and 12.8%, respectively. Some analytes were excluded from the evaluation set due to considerations of volatility (boiling point <400 °C), charge and excessive retention on the column leading to elution outside the eluent range covered by the model. The two-part response model can be used to describe the relationship between response and analyte concentration and also to offer a correction for the non-linear detector response obtained with gradient HPLC for analytes which conform to the model, to provide insight into the factors affecting the CoronaCAD response for different analytes, and also as a means for accurately determining the concentration of unknown compounds when individual standards are not available for calibration.

Development and Validation of a RP-HPLC Method for the Estimation of Netilmicin Sulfate and its Related Substances using Charged Aerosol Detection

Netilmicin is a semi-synthetic aminoglycoside antibiotic used against a broad spectrum of Gram-negative bacteria. A reversed-phase high-performance liquid chromatographic method has been developed to determine the composition of netilmicin sulfate and to estimate its related substances without pre- or post-column derivatization. A UV detector cannot be used to detect low levels of known and unknown related substances of netilmicin, as it has only a weak UV chromophore. A charged aerosol detector was used instead to obtain the high sensitivity that was necessary for the intended purpose of the method. This method can separate all related substances of netilmicin. A (10 cm x 4.6 mm) pentafluorophenyl high-performance liquid chromatographic column from Restek was used with a mobile phase consisting of (A) pentafluoropropionic acid-water-acetonitrile (0.1:96:4, v/v/v) and (B) trifluoroacetic acid-water-acetonitrile (1:96:4, v/v/v).

Simultaneous determination of positive and negative pharmaceutical counterions using mixed-mode chromatography coupled with charged aerosol detector

A mixed-mode chromatography coupled with charged aerosol detector (CAD) method was developed in this work to simultaneously determine pharmaceutical counterions including both inorganic ions and organic ions in the forms of cations and anions. 25 commonly used pharmaceutical ions were studied and simultaneously separated within 20 min by this single method. A silica based mixed-mode column with reversed-phase/cation-exchange/anion-exchange modes was used. It provides reversed-phase, strong cation-exchange and weak anion-exchange properties at the same time. It also provides the HILIC behavior at high percentage of organic solvent. The effects of mobile-phase organic strength, buffer ions, ionic strength, pH and column temperature have been investigated to optimize the method as well as to understand the retention and separation mechanisms. Conventional HPLC system was used and no special chromatography system is needed. The presented method has been employed successfully for screening and quantitative analysis of counterions, unknown ionic impurities and salts in active pharmaceutical ingredients and in process control samples with excellent accuracy, precision and sensitivity. This method provides a simple, fast and generic approach to speed up pharmaceutical research and development process and enhance lab efficiency. The similar methodologies can be applied to other ion analysis.