The United States Pharmacopeia (USP) has updated General Chapter <591> «Zinc Determination» to include ion chromatography (IC) as a method to quantitatively determine zinc, including zinc oxide, in drug substance and drug product monographs. Zinc oxide is one of the active ingredients approved by the U.S. FDA for sunscreens. Zinc oxide determinations in sunscreens were previously demonstrated in Thermo Scientific Application Note 72680. This method here demonstrates the application using a Thermo Scientific™ Dionex™ Inuvion™ Core IC system, a new integrated, single channel, compact IC.
This application proof note demonstrates a determination of citrate and phosphate in pharmaceutical formulations using the method published in Thermo Fisher Scientific Application Note 164. In this proof note, the method is performed using Inuvion IC system.
Sodium bicarbonate, or sodium hydrogen carbonate, is used in a wide variety of pharmaceutical and food products, and ammonia is one of the impurities that is assayed. In this application proof note, the performance of the easy-to-use, compact Thermo Scientific Dionex Inuvion ion chromatography system was evaluated for this application.
This application developed a new method for the determination of fluoride in oral rinse products using carbonate/bicarbonate eluent. The method demonstrates excellent accuracy and precision for fluoride in commercially available oral rinse samples and, therefore, can be used as a quality control method by oral rinse manufacturers.
This application modified the existing USP Sodium Fluoride monograph method for determining fluoride in oral rinse products with a modern, high-performance anion-exchange column optimized for using carbonate-based eluents, as suggested by the USP. The method demonstrates excellent accuracy and precision for fluoride in commercially available oral rinse sample, and, therefore, can be used as a quality control method by oral rinse manufacturers.
In this application note, we demonstrate a workflow combining the chemical release of O-linked glycans, followed by sample cleanup and analysis by HPAE-PAD hyphenated to a Q Exactive HF mass spectrometer. The Dionex CarboPac PA300-4μm column uses a novel column technology that enables the simultaneous separation of neutral and charged glycans without the need of derivatization. High resolution accurate mass MS data and tandem MS/MS spectra with diagnostic fragment ions provide highly reliable structural annotations of heterogenous glycans.
This application note describes a method for nucleotide sugar determination using anion-exchange chromatography. This method uses a Thermo Scientific™ Dionex™ CarboPac™ PA1 column and manually prepared eluents for separation. Using a this column, a 34 min method that resolved seven analytes was designed.
The goal of this work is to design an ion chromatography (IC) method for the determination of guanidine. A method that uses a Thermo Scientific™ Dionex™ IonPac™ CS20 cation-exchange column, electrolytically generated MSA eluent, and suppressed conductivity detection was developed to determine guanidine in pharmaceutical formulations. The method proposed in this application note was validated following the guidelines outlined in USP General Chapter <1225>, Validation of Compendial Procedures to meet the requirements of a validated guanidine assay.
The goal of this work is to design an ion chromatography method that uses a Dionex IonPac AS28-Fast-4μm anion exchange column, electrolytically generated KOH eluent, and suppressed conductivity detection to determine phosphite and phosphate in ibandronate sodium. The method proposed in this application note was validated following the guidelines outlined in USP General Chapter <1225>, Validation of Compendial Procedures to meet the requirements specified for phosphite and phosphate quantification prescribed in the proposed USP Ibandronate Sodium monograph.
This work describes an HPAE-PAD method for the determination of ribitol in Hib capsular polysaccharide acid hydrolysis samples. Separation of ribitol from TFA acid hydrolysis products was achieved under isocratic elution conditions using a Thermo Scientific™ Dionex™ CarboPac™ MA1 column. The method proposed here was validated for performance with respect to linearity, precision, sensitivity, and accuracy. The acid hydrolysis samples were also tested using a palladium hydrogen reference electrode.
KDO (3-deoxy-D-manno-oct-2-ulosonic acid) is an eight-carbon sugar mostly confined to Gram-negative bacteria. KDO can be used to quantify the amount of a known lipopolysaccharide (LPS) (the moles of KDO per mole of liposaccharide must be known). The hydrolysis conditions were optimized for the release of KDO from an LPS and a method for its quantitation has been developed using HPAE-PAD. In this application note, we develop, optimize, and validate a HPAE-PAD method for the determination of KDO.
This application note describes a convenient, fast, and sensitive method that uses a Thermo Scientific™ Dionex™ CarboPac™ PA300 column and electrolytic eluent generation. The Dionex CarboPac PA300-4μm column is packed with a new 4 μm particle size supermacroporous resin to achieve high efficiency and high-resolution separations. Using this column, a 16.8 min method that resolved six analytes, including glycol and other carbohydrates, was designed. Results for method linearity, robustness, and accuracy for glycol and carbohydrate quantification in OTC samples are discussed here.
Tromethamine is commonly used as a buffering agent, alkalizer, and emulsifying agent in pharmaceutical and cosmetic preparations, and as a counterion for acidic drug substances. This work describes an IC method that uses a Thermo Scientific™ Dionex™ IonPac™ CS20 cation exchange column, electrolytically generated MSA eluent, and suppressed conductivity detection to determine tromethamine in pharmaceutical formulations.
We demonstrate a workflow combining chemical release of O-linked glycans, followed by sample cleanup and analysis by HPAE-PAD hyphenated to a Q Exactive HF Hybrid Quadrupole-Orbitrap MS. A recently introduced Dionex CarboPac PA300-4μm column, enables simultaneous separation of neutral and charged glycans without the need for derivatization. High-resolution MS data and MS/MS spectra with diagnostic fragment ions provide reliable structural annotations of heterogeneous glycans. We applied HPAE-PAD/MS to analyze O-glycan structures released from four different glycoproteins.
This record has QAR conditions and results for the 2 mm Dionex IonPac CS19-4µm column. The column must be pre-conditioned offline with acetonitrile and methanesulfonic acid (MSA). See instructions in the column manual (attached). The column is optimized for small, hydrophilic amines such as ethanolamines, methylamines, ethylamines, as well as the biogenic amines.
This application note demonstrates that the AU180 method for sialic acid determination of glycoproteins could be successfully modified for use with a Dionex CarboPac PA20-1 mm column and HPAE-PAD in dual eluent generation cartridge mode.
This technical note (TN) describes how to develop an ion chromatography method to determine an amine in pharmaceutical samples. Two methods were developed in this TN. Method choice depends on the nature of the sample. This record is for the method that uses the Dionex IonPac CS19 column .
This technical note (TN) describes how to develop an ion chromatography method to determine an amine in pharmaceutical samples. Two methods were developed in this TN. Method choice depends on the nature of the sample. This record is for the method that uses the Dionex IonPac CS16 column .
A method was developed for the determination of nitrite in pharmaceuticals by coupling IC with UV absorbance detection. The LOD of nitrite in a pharmaceutical sample is 0.918 ppm (μg/g API). The method is accurate and precise due to the high reproducibility of the Reagent-Free ion chromatography system. This method should be applicable to the determination of nitrite throughout the manufacturing process of a drug product to assess the likelihood of nitrosamine formation.
In this work, we evaluated the IC method in the proposed USP monograph for the MgO assay and Limit of Calcium test. Magnesium and calcium ions were separated on a Dionex IonPac CS16 column set followed by suppressed conductivity detection. Key performance parameters were evaluated, including separation, system suitability, linearity, limit of detection, and precision. MgO tablets and capsules were analyzed and evaluated in accordance with the proposed USP monographs. The percentage of MgO and the limit of calcium results were compared with the proposed USP monographs’ acceptance criteria.
This record describes the QAR analysis for a Thermo Scientific Dionex CarboPac PA300 column and provides an eWorkflow for running the analysis..
Boric acid and borates (e.g., metaborate, borate, and tetraborate) are added to some cosmetics as a preservative and antibacterial agent. But once inhaled or absorbed through skin cuts, boric acid and borates can cause intoxication. An IC method that has been applied to the sensitive determination of borate (boric acid) in water samples is applied to the determination of borate in cosmetics. Borate is separated using 3 mM MSA 60 mM mannitol eluent on a Dionex ICE-borate column and detected by suppressed conductivity using external chemical regenerant.
We evaluated the proposed USP Busulfan monograph method for the “Limit of Methanesulfonic Acid”. MSA was separated on a Dionex IonPac AS11-HC column set followed by suppressed conductivity detection. The method was further evaluated with a slight modification to the sample preparation method. The busulfan sample was prepared in cold 30:70 acetonitrile:water (30% MeCN) instead of water. We also set the autosampler temperature set to 8 °C instead of 25 °C. Key performance parameters were evaluated including separation, system suitability, linearity, limit of detection, and precision.
We developed a sensitive and robust ion chromatography (IC)-based test for the determination of ammonia impurity in potassium bitartrate to modernize the test in the United States Pharmacopeia’s (USP) Potassium Bitartrate monograph. Our IC method offers a significant improvement to the current impurity test in the USP monograph. The 5 min method uses a Thermo Scientific™ Dionex™ IonPac™ CS16 high capacity column, an electrolytically generated methanesulfonic acid (MSA) eluent, and suppressed conductivity detection on a Reagent-Free™ Ion Chromatography (RFIC™) system.
This application note illustrates the use of an easy-to-implement IC-MS method for the determination of aliphatic and unsaturated organic acids in pharmaceutical solutions. This workflow employs both suppressed conductivity and MS detection to increase the information available from each sample. Samples were separated on a high-resolution Dionex™ IonPac™ AS11-HC-4μm column set using a Dionex™ Integrion™ HPIC™ system with suppressed conductivity detection. The sequential MS detection was achieved using a Thermo Scientific™ ISQ™ EC single quadrupole mass spectrometer.
In this application note, the nitrite determination described in the USP monograph was evaluated with a Thermo Scientific Dionex IonPac AS15 column using an IC system. The method and conditions were exactly as described in the USP Dalteparin Sodium monograph. Key performance parameters were evaluated including the system suitability separation, linearity, limits of detection, accuracy, and robustness. One Dalteparin Sodium sample was analyzed. The percentage of nitrite result was compared with the USP acceptance criterium.
This application note describes an IC-based method that uses a Thermo Scientific™ Dionex™ IonPac™ AS20 anion exchange column, an electrolytically generated potassium hydroxide eluent, and suppressed conductivity detection to determine oxalate in cromolyn sodium. The method proposed in this application note was validated following the guidelines outlined in USP General Chapter <1225>, Validation of Compendial Procedures.
This application note describes an IC-based method that uses a Thermo Scientific™ Dionex™ IonPac™ CS5A cation-exchange column, pyridine-2,6-dicarboxylic acid (PDCA) eluent, post-column reaction with 4-(2-pyridylazo)resorcinol (PAR), and absorbance detection to determine copper in cupric chloride. The Dionex IonPac CS5A high-resolution column is designed for fast and accurate determination of the transition and lanthanide metals in a variety of samples. The method proposed here was validated following the guidelines outlined in USP General Chapter <1225>, Validation of Compendial Procedures.
We recently introduced a PdH RE for HPAE-PAD carbohydrate determinations. In this work, we demonstrate two popular carbohydrate applications (honey sugars and glycoprotein monosaccharides) using a PdH RE and compare the results to using an Ag/AgCl RE. Our preliminary data indicate that the PdH RE provides comparable results to the standard Ag/AgCl RE for these two applications that use hydroxide eluents.
The methods proposed here use a Thermo Scientific™ Dionex™ CarboPac™ PA200 column for glycan separation by HPAE. After the separation the glycans pass through a desalting device and in to the mass spectrometer. Fragmentation of glycans in the negative mode by higher-energy collisional dissociation (HCD) provides information-rich MS2 spectra which enables glycan structure annotation. This allowed correlation of glycan structure with observed elution behavior. Here we show that in some cases, changed elution conditions can be used to resolve different glycan structures.
Cyclodextrins (CDs) have been used as complexing agents to increase water solubility of poorly soluble drugs and to increase their bioavailability.. The presence of oxidizable hydroxyl groups makes CDs well suited for determination by HPAE-PAD. Here, we used Thermo Scientific Dionex CarboPac PA200 Column to evaluate the “Limit of β-CD" according to the USP Monograph. Key performance parameters were evaluated and compared including system suitability, linearity, limits of detection, and precision. The application of the Dionex CarboPac PA200 column to separating other CDs was also examined.
We developed an ion chromatography (IC) method to replace the titrimetric assay for potassium bitartrate in the United States Pharmacopeia (USP) Potassium Bitartrate monograph. Our IC method offers a significant improvement to the existing assay in the USP monograph. It is a 5 min method that uses a Dionex™ IonPac™ AS20 ion-exchange column, an electrolytically generated KOH eluent, and suppressed conductivity detection on a Thermo Scientific™ Dionex™ Reagent-Free™ Ion Chromatography (RFIC™) system supported by regulatory compliant Chromeleon™ Chromatography Data System (CDS) software.
Before running any samples, Thermo Scientific recommends that you first confirm the performance of the column by reproducing the lot validation report chromatogram shipped with column. Compare your results with the one reported in the quality assurance report. At least three injections should be made. This record provides an eWorkflow for executing the QAR method on an ICS-5000+ system. An ICS-6000 can be used for this application.
Sodium monofluorophosphate (MFP) is approved by the FDA as an anti-caries agent in toothpastes and mouth rinses. MFP can hydrolyze to free fluoride and phosphate during storage. It is important to determine fluoride and MFP in MFP solid and in toothpastes to evaluate API and drug product quality and stability. An IC method has been proposed to replace existing titration-based assays in the MFP monograph. The IC method used an anion-exchange column with suppressed conductivity detection. This application validated the IC method and demonstrated using it to determine MFP in a toothpaste.
Glycoprotein characterization and glycosylation profiling are important tasks in the development and production of biopharmaceutical proteins. This application note demonstrates Dual EGC capability and performance for profiling N-linked oligosaccharides released from glycoproteins using high-performance anion exchange chromatography with pulsed amperometric detection (HPAE-PAD). This record shows the separation of N-linked glycans from human acid-1 glycoprotein.
Glycoprotein characterization and glycosylation profiling are important tasks in the development and production of biopharmaceutical proteins. This application note demonstrates Dual EGC capability and performance for profiling N-linked oligosaccharides released from glycoproteins using high-performance anion exchange chromatography with pulsed amperometric detection (HPAE-PAD). This record shows the separation for IgG N-linked glycans.
Due to the very polar and strong chelating characteristics of bisphosphonates, their analysis represents a real analytical challenge. Here separation of N-containing bisphosphonates and three other bisphosphonates was achieved without derivatization. After an anion-exchange separation, the analytes were passed through an anion suppressor before detection by suppressed conductivity and single quadrupole mass spectrometry in Selected Ion Monitoring mode without the aid of a desolvation agent at any step of separation or electrospray ionization.
Here we demonstrate that a Dionex IonPac AS15-5μm column can be used to analyze tooth gel samples for the content of fluoride. Fluoride is separated from other anions using KOH eluent generated electrolytically with a EGC III KOH cartridge and detected by suppressed conductivity. Method performance is evaluated according to the system suitability requirements described in the proposed United States Pharmacopeia (USP) monograph revision for the assay and the acceptance criterion set for sodium fluoride content in a tooth gel sample.
This application note demonstrates that spectinomycin dihydrochloride and related impurities can be separated with a Dionex IonPac AmG-3µm C18 column using a simple eluent method (0.1M TFA) compared to the method in the EP monograph. The separation, repeatability, and sensitivity of this method were found to meet or exceed the current EP Spectinomycin Dihydrochloride monograph performance requirements. This method is reliable and can be used for the routine monitoring of spectinomycin.
This application update demonstrates improved resolution of sialylated N-glycans on Thermo Scientific™ Dionex™ CarboPac™ PA200 columns. Starting with a recently described method, changes to elution conditions were tested to improve resolution of sialylated N-glycans enzymatically released from four different glycoproteins. Separations were first evaluated on the analytical format (3 × 250 mm column). Next, the possibility that a shorter column, such as a guard column, would allow significantly improved throughput was evaluated. Finally, a narrow bore format 1 × 250 mm column was tested.
This application note demonstrated that the ChP Etimicin Sulfate monograph etimicin assay method and the method for organic impurities could be successfully executed with a Dionex IonPac AmG-3µm C18 column on an ICS-5000+ system. The separation, linearity, reproducibility, and sensitivity were found to meet or exceed the current ChP Etimicin Sulfate monograph performance requirements. This method is reliable and can be used for the routine monitoring of etimicin.
In this application note, we evaluated the USP monograph method. Betadex was separated on a Dionex IonPac AS11 column followed by PAD detection using the specified 3-potential waveform with a gold conventional working electrode. The method was further evaluated with the 4-potential waveform recommended for carbohydrates and a gold disposable working electrode. In this additional evaluation only the waveform was changed compared to the monograph method. Key performance parameters were evaluated including separation, system suitability, linearity, limits of detection, and precision.
This study demonstrates use of a commercially available glycopeptide standard as a control to determine glycoprotein hydrolysis reaction efficiency. Because the monosaccharide composition of glycopeptide is known, hydrolysis efficiency under a set of conditions can be easily calculated. The monosaccharide yield for the glycopeptide was compared with that of two glycoproteins under identical conditions. This comparison revealed efficient hydrolysis using the conditions tested here. Minimally the glycopeptide standard serves as a positive control for glycoprotein monosaccharide determination.
Before running any samples, Thermo Scientific recommends that you first confirm the performance of the column by reproducing the lot validation report chromatogram shipped with column. Compare your results with the one reported in the quality assurance report. At least three injections should be made. This record provides an eWorkflow for executing the QAR method on an ICS-6000 system using Dual EGC mode.
Before running any samples, Thermo Scientific recommends that you first confirm the performance of the column by reproducing the lot validation report chromatogram shipped with column. Compare your results with the one reported in the quality assurance report. This record provides an eWorkflow for executing the QAR method on an HPIC system. See column manual for detail.
This application update demonstrates that gentamicin sulfate and related impurities can be separated quickly compared to the method in the United States Pharmacopeia (USP) Gentamicin Sulfate monograph using a Dionex IonPac AmG-3µm C18 column with 100 mM TFA (trifluoroacetic acid) with 2% acetonitrile as the eluent . (Method B in AU 72648)). This method meets or exceeds the performance criteria for Content of Gentamicins in the USP Gentamicin Sulfate monograph.
This application update demonstrates that gentamicin sulfate and related impurities can be separated with a Dionex IonPac AmG-3µm C18 column using a simple eluent method (100 mM TFA (trifluoroacetic acid) and no organic solvent. (Method A in AU 72648)). This method meets or exceeds the performance criteria for Content of Gentamicins in the United States Pharmacopeia (USP) Gentamicin Sulfate monograph.
This application note demonstrates that the USP Gentamicin Sulfate monograph Content of Gentamicins method and the USP in-process revision Gentamicin Sulfate monograph method for organic impurities method can be successfully executed with a Dionex IonPac AmG-3µm C18 column using either the 4- or 3-potential carbohydrate waveform. The separation, linearity, reproducibility, and sensitivity were found to meet or exceed the current USP/EP Gentamicin Sulfate monograph performance requirements. This method is reliable and can be used for the routine monitoring of gentamicin.
Zinc oxide is approved by the FDA for use in sunscreens. An ion chromatography (IC) method has been proposed to replace existing titration-based assays in the monograph. The Zinc Oxide content was determined using a IC system with a cation-exchange column. The determination of zinc is achieved by the formation of anionic complexes with a eluent that contains pyridine-2,6-dicarboxylic acid (PDCA), post column derivative with 4-(2-pyridylazo)resorcinol (PAR), and absorbance detection at 530 nm. The method was validated following USP guidelines and demonstrated with different sunscreens.