Sertraline Tablet Analyzed with HPLC – AppNote

Retention of Hydrophobic and Hydrophilic Analytes with Zoloft® Sertraline has a logP of 2.9, but retains well when using Aqueous Normal Phase (ANP) HPLC. A polar test solute, Phenylglycine, was added to demonstrate how both nonpolar and polar compounds can be retained in the same Method. This would be important when looking for impurities or […]

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Shikimic Acid Analysis by LCMS- AppNote

Ingredient for Production of Oseltamivir The method shown in this application note assures both high specificity and high sensitivity for Shikimic Acid, a synthetic ingredient for the production of the pharmaceutical Oseltamivir marketed as Tamiflu®. Using this LCMS friendly method allows for a 10-100 fold increase in sensitivity over some other methods. Also note that […]

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Shikimic Acid in Red Wine Analyzed with LCMS – AppNote

Red Wine Varietal Determined by Quantity of Shikimic Acid This Application Note presents a Method for the analysis of Shikimic Acid in Red Wine. The main advantage of using this Method is its high Specificity (mass accuracy) of this compound, the short analysis time, fast Equilibration time between runs thus more throughput, and high Repeatability […]

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Sorbic and Tartaric Acids in Wine Analyzed by LCMS – AppNote

Minimal Sample Preparation This method doesn’t require complex preparation or extraction of the wine samples. It allows detection of both Acids in a short gradient run, with rapid equilibration of the column after each run. Calibration curves were constructed in the ranges 1–300ppm and 100–3000ppm for Sorbic Acid and Tartaric Acid respectively. Correlation coefficients for […]

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Streptomycin Analyzed with HPLC – AppNote

Retention of a Highly Polar Antibiotic Streptomycin is very polar and difficult to retain with Reversed Phase methods using conventional Columns. In this Method, retention is achieved with Separation of the Streptomycin from two impurities found in the Standard Solution. Furthermore, the numerous amine groups do not cause tailing of Peak shape as with standard […]

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Succinic Acid Analyzed with LCMS – AppNote

Isocratic or Gradient The analysis of Succinic Acid is mostly performed by Gas Chromatography (GC), Enzymatic Assays, Ion-Exclusion Chromatography (IEC) or by Reversed Phase HPLC. However, these techniques are time consuming and analysis time is very often longer than 1 hour. In this Application Note, a rapid, cost effective screening Method for the detection of […]

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Sugar Nucleotides Analyzed with LCMS/MS – AppNote

Separating UDP and CDP Sugars with ANP and Increased Sensitivity This Method can be used to analyze UDP and CDP Sugars using UDP-Hexanolamine (a metabolite) as an Internal standard. The Sugar Nucleotides used in this Application Note are a mixture of compounds that occur in plants and their structure is proprietary. A potentially powerful tool […]

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Sugars, Sugar Alcohols & Amino Acids Separation with LCMS – AppNote

Glucose, Mannitol, Aspartic Acid, Alanine and Arginine Metabolomics analysis encompasses a vast array of molecules with disparate hydrophobic/hydrophilic properties. Many  of these analytes are in fact quite polar and problematic to retain by conventional Reversed Phase Chromatography. The Cogent Diamond Hydride Column is quite well-suited to their separation. This application demonstrates this property through the […]

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Sumatriptan as an API Analyzed with LCMS – AppNote

Analysis of Imitrex® API from Tablet Formulations This AppNote Method for Analysis of a challenging compound (Sumatriptan) has several advantages over other Methods. This Analysis can be performed using a higher concentration of organic solvent, which is much more suitable for electrospray MS Detection. This AppNote provides robust and repeatable (%RSD 0.1 and lower) results. […]

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Tandem LCMS approach to Metabolite Profiling – Poster

Hydrophilic and Hydrophobic Molecules In a Single Line Injection Presented at the Metabolomics Society meeting in Amsterdam, June 2010; below is the introduction from the poster or you can download and view the entire poster by clicking on the link below. INTRODUCTION: A tandem liquid chromatography (LC) setup has been developed to analyze a larger […]

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Tapentadol HCl Tablet Analyzed with HPLC – AppNote

Assay Method for the Analgesic Nucynta Tapentadol can be a problematic compound for HPLC analysis due to the Amine functional group. Tertiary amines are often particularly difficult to obtain a good peak shape using Reversed Phase methods. Peak tailing has been reported in several published papers in the literature. With this Method, a sharp Peak […]

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Taurine Analyzed with LCMS – AppNote

Retention of a Polar, Sulfonate Compound As a highly polar compound, Taurine is often difficult to retain by Reversed Phase Chromatography. The Peak Shape and Retention obtained with this Aqueous Normal Phase (ANP) LCMS Method is greatly improved and is suitable as a starting point for further analyses of Taurine, in a variety of Samples […]

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Taurine with HPLC & Refractive Index – AppNote

Retention of Highly Polar Compound The food and beverage industry may need reliable methods for assay for Taurine in their products. However, its analysis is complicated by the lack of chromophores in its structure. LC-MS can be used instead, but many QC labs may not have this instrumentation or might prefer a more simple detection […]

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Terbutaline and Salbutamol Analyzed with HPLC – AppNote

Separation of ß2-Adrenergic Receptor Agonists This Method shows how two structurally similar compounds can be readily separated. This is an important characteristic in many types of HPLC Methods since typical separations often involve impurities, degradants, metabolites, and other groups of compounds that have subtle structural differences. These two compounds for instance were found to coelute […]

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Tip for Promoting Analyte Ionization with LCMS – AppNote

Analyte Ionization using L-Tyrosine Figures A and B show the Peak for L-Tyrosine obtained with two LCMS Methods. The analysis shown in Figure B was performed with a stronger ANP Solvent (DI Water) as component A of the Mobile Phase. The obtained Retention time was lower than in Figure A, as was the compound ionization. […]

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Tizanidine HCl HPLC Method Transferred to Near UHPLC – AppNote

Use of Near UHPLC Column for Improved Results This challenging compound has several amine groups and can be problematic to analyze with HPLC. Here a good peak shape is obtained using the Cogent Diamond Hydride 2.o Column. The efficiency is higher on the 2.2µm phase compared to a standard 4µm Column, leading to increased sensitivity. […]

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Tizanidine HCl Tablet Analyzed with HPLC – AppNote

Separation Method for Tizanidine Compatible with LC-MS Tizanidine has numerous amine functional groups and can be a challenge for analysis with HPLC. The USP method uses Phosphate in the Mobile Phase which is not compatible with LC-MS. This Method however uses Formic Acid as the Mobile Phase additive and produces a sharp, symmetrical peak. The […]

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Tizanidine HCl Tablet with Precision – AppNote

Excellent Run to Run Precision Tizanidine has several amine groups which can present problems to chromatographers. Residual silanols on the surface of ordinary Silica-based Columns can electrostatically interact with these groups, causing peak tailing. The Cogent Amide Column, used in the Application Note is virtually free of residual silanols and therefore a symmetrical peak shape […]

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Tramadol Separation with Enhanced LCMS – AppNote

LC-MS Compatible HPLC Method for Tramadol  Tramadol can be retained by Reversed Phase HPLC  but some reported methods require Phosphate buffer additives in the Mobile Phase, which means incompatibility with LC-MS. In contrast, this Method uses Formic Acid as the Mobile Phase additive. An advantage this method has is that it produces enhanced signal-to-noise compared […]

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Trans-3-Hydroxycinnamic Acid in Human Urine with LCMS – AppNote

Separation of Antioxidants with HPLC with LCMS Polar metabolites are reproducibly retained using an MS friendly Mobile Phase in Aqueous Normal Phase (ANP) HPLC. The sample preparation required is very easy and cost effective compared to previous, older Methods. The sample is diluted with the Reversed Phase Solvents and ready to inject in 5-10 minutes. […]

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Triamterene Analyzed with HPLC – AppNote

Symmetrical Peak Shape without Ion Pair Agents Triamterene has numerous amine functional groups and therefore tails readily in Reversed Phase Methods if ion pair agents are not used in the Mobile Phase. However, these additives not only contribute to a less Robust Method in terms of Reproducibility of analyte Retention, but are also incompatible with […]

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Trimethobenzamide Analyzed with HPLC – AppNote

Separation of Trimethobenzamide with a Simple Isocratic Method  The USP Assay Method of Trimethobenzamide is performed by titration. This alternative HPLC approach is fast, easy, and reliable. It has many advantages over the titration Method, including ease of automation and less operator-dependent results. This simple isocratic Method provides sufficient Retention of the API to allow […]

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Tropicamide and Homatropine Analyzed with HPLC – AppNote

Separation of Anticholinergic Mydriatic Agents In this Method, Tropicamide and Homatropine are separated with a simple Gradient. Homatropine is significantly hydrophobic yet still retains well. Furthermore, the Separation between these two solutes is good, illustrating the Selectivity capabilities of this Method. Data is shown for two lots of Columns in order to demonstrate the robustness […]

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Tryptamine Analyzed with HPLC – AppNote

Simple and Fast Analyses of an Important Biogenic Amine Using this Method to analyze Tryptamine produces excellent Peak Shape and Reproducible Retention Times as shown. It is very polar and very difficult to Retain when using standard C18 Columns. The baseline in this Chromatogram appears to be noisy but it is due the low UV […]

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Tyramine, N-Methyltyramine & Synephrine Analyzed with LCMS – AppNote

Adrenergic Amines found in Citrus Fruit Juice with LCMS This Method, developed for the analysis of Tangerine Juice is Sensitive, Fast, Simple and Reproducible when using a Mass Spectrometer. The resulting Sample and diluent, after centrifugation, is fully compatible with the Mobile Phase (listed below), which allows direct injection with no further Sample Prep being […]

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UDP-Glucose Analyzed with LCMS – AppNote

Analyzing a Nucleotide Sugar with UDP-Hexanolamine as an Internal Standard This Method is rapid and simple and it can also be used in measuring UDP Glucose. The advantages of using this Method to assay Nucleotide Sugars such as this one are the short Separation Time, long term Column Stability and rapid Equilibration time between a […]

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Unique Method Development Strategy for Polar Compounds – AppNote

Anatoxin-a Analyzed with LCMS Figures below illustrate an example of the work flow in developing a Method for the Analysis of a Polar Compound using Cogent Diamond Hydride™ Columns and the impact of Aqueous Normal Phase HPLC. The simplified steps of Method Development are as follows: —A. Injection of the Sample at 50:50 Solvent A […]

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Urea Analyzed with HPLC – AppNote

Retention of Urea, a Highly Polar Compound Urea is very difficult to retain by conventional HPLC methods. It is highly polar and therefore shows little or no Reversed Phase retention. However, this Method shows how it can be readily retained past the solvent front using a simple isocratic Mobile Phase. Furthermore, the Peak shape for […]

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Uric Acid (UA) Metabolites Analyzed by LCMS – AppNote

Allantoin and 6-Aminouracil The presented method for analysis of UA metabolites in human Urine is very simple and does not require Mobile Phase additives or pre-Column derivatization. The peaks are symmetrical and the MS signal is not diminished as is the case when Ammonium Formate or Acetate is used as the Mobile Phase additive. The […]

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Uric acid and Metabolites Analyzed by LCMS – AppNote

LCMS Method for Allantoin, 6-Aminouracil, and Uric Acid (UA) The presented method for analysis of UA and its highly polar metabolites is simple and does not require Mobile Phase additives or pre-column derivatization. The Peaks are symmetrical, and the MS signal is not diminished, as is the case when Ammonium Formate or Acetate is used […]

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Valaciclovir & Degradants Analyzed with LCMS – AppNote

Separation of Valaciclovir from two Degradants The anti-viral Herpes drug Valaciclovir and its degradation products are analyzed using Aqueous Normal Phase HPLC with MS detection. The Method development was completed using UV detection, where excellent Separation between the Prodrug Valaciclovir and its active converted degradant, Acyclovir, was observed. The Method transfer from UV to MS […]

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Valaciclovir Analyzed with HPLC – AppNote

Separation of Prodrug from a Degradant in Forced Degradation of Valtrex® Tablets In this Application Note, the anti-viral Herpes drug Valaciclovir and its main acid degradant are well separated (Figure A). Valaciclovir is a prodrug and the degradant observed here is believed to be the active form, Acyclovir. Both compounds did not retain very strongly […]

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Vancomycin Analyzed with HPLC- AppNote

Retention of a Highly Polar Antibiotic As a highly polar compound, Vancomycin is difficult to retain with Reversed Phase Methods. In this Method the compound retains very well as illustrated in the Figure. In addition, the repeatability of the analysis, which is demonstrated by the overlay of five consecutive runs, is excellent. The Mobile Phase […]

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Vancomycin HPLC Method Transferred – AppNote

Benefits of using 2.2 µm Column for Vancomycin Analysis The highly polar antibiotic Vancomycin can be readily analyzed with either the standard 4µm Cogent Diamond Hydride or the Cogent Diamond Hydride 2.o Column. Notably higher efficiency is obtained on the smaller particle size Column. This Method is easy to perform and is LCMS compatible. Three […]

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Vardenafil (Levitra®) Analyzed with HPLC – AppNote

LC-MS Compatible Assay Method Vardenafil in a tablet formulation can be readily assayed with this LC-MS compatible gradient method. The peak tailing factor was close to unity. The compound has several amine groups which can produce tailing with ordinary HPLC Columns that have a number of surface Silanols. The MS-compatible Mobile Phase means that the […]

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Verapamil and Impurity Analyzed with HPLC – AppNote

Alternative to USP Method with Improved Peak Efficiency  The USP impurities Method for Verapamil uses an Acetate Buffer with the ion pair agent 2-Aminoheptane to reduce Peak Tailing of the API and its Impurity. This Application Note uses an LCMS compatible Mobile Phase and produces Symmetrical Peaks for both Compounds. Excellent Resolution was also obtained […]

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Warfarin, Hydroxybupropion, and Codeine by LCMS – AppNote

Analysis in Spiked Blood Samples In this application, three pharmaceuticals/metabolites are separated using the Cogent Diol 2.o Column. The blood sample illustrates the suitability of the Column for analysis of more complex matrices. Use of Acetone in the Mobile Phase was found to be appropriate for this LC-MS method. As a less expensive and less […]

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Water Soluble Vitamin Tablet Analysis with HPLC – AppNote

Separation of Water Soluble Vitamins without Ion Pair Agents This AppNote illustrates the Separation of four Hydrophilic, B Vitamins which were extracted from a commercially available tablet formulation. The data shows that these Water soluble B-Vitamins are well Separated from each other and from any Matrix Peaks. The main advantage of this Method over traditional […]

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Xanthine, Uric Acid & Hypoxanthine Analyzed with LCMS – AppNote

Separation of Biomarkers found in Human Fluids A simple Method was developed for the determination of Xanthine (X), Uric Acid (UA), and Hypoxanthine (HX) at concentrations in human urine (can be used for human serum) to support pharmacodynamic (PD) studies of a novel Xanthine Oxidase inhibitor during its clinical development. PD Biomarkers (UA, X, and […]

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Zinc-EDTA Complex – AppNote

No Ion Pair Agents Needed Using conventional analytical methods, retention of metal–EDTA complexes is accomplished using Ion Pair Reversed Phase Chromatography. However, the Ion Pair Agents used in the Mobile Phase are not compatible with Mass Spectrometry. In this AppNote, only Formic Acid is needed in the Mobile Phase in order to obtain retention of […]

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