Metaldehyde in Slug Pellets Analyzed by LCMS – AppNote

Pesticide Formulation As Metaldehyde is a non-UV absorbing compound, other detection methods needed to be investigated besides conventional UV-HPLC. LC-MS was found to be a well-suited analysis by searching for the EIC corresponding to the [M+H]+ ion. Good retention and peak shape were observed for this analyte using the Cogent Bidentate C18 2.o™ Column. Peak: […]

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Molnupiravir and Favipiravir Analyzed with HPLC – AppNote

Oral Anti-Viral Medications – Easy Reversed Phase Method  Molnupiravir and Favipiravir, new anti-viral drugs were analyzed with HPLC using a simple Mobile Phase. As shown in the 10 injection overlay in the chromatogram below, the Separation, Peak Shapes and Repeatability are very good (%RSD ≤ 0.2).   Peaks: 1. Favipiravir 2. Molnupiravir  Method Conditions: Column: Cogent […]

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Mometasone Furoate in Topical Cream Analyzed with HPLC – AppNote

Separation of Prodrug from Acid Hydrolysis Products This Method shows the Separation of an ester Prodrug, Mometasone Furoate, from another compound formed during acid hydrolysis. This Peak is quite possibly the active form, Mometasone. The two Chromatograms show the non-degraded extract (Figure A) and an acid hydrolysis extract (Figure B). Hydrolysis of the ester group […]

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Monoclonal Anti-A1-Glycoprotein – AppNote

Glycoprotein Analysis  For the last few decades Glycoproteins have been a subject of interest to biochemists and biologists in many fields. These proteins are found in plasma and other biological fluids and they serve many functions in nearly every physiological process of living organisms. These macromolecules consist of a peptide chain and one or more […]

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Morphine Sulfate Tablet HPLC Method Transferred to Near UHPLC – AppNote

Standard 4um Particle Size Transferred to 2.o™ This application illustrates how methods developed using the 4µm Cogent Diamond Hydride Columns may be adapted for Cogent Diamond Hydride 2.o™ phases. Morphine shows slightly higher retention on the 2.o™ Column (average 7.311 min vs. 5.588 min). The efficiency is almost twice as high when using the smaller […]

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Morphine, Hydromorphone and 6-MAM in Plasma Analyzed with LCMS – AppNote

Morphine, Hydromorphone and 6-Monoacetylmorphine (6-MAM) in Plasma This AppNote presents a Method that could be used for a routine analysis of Plasma Samples or (whole blood samples after changing the extraction procedure) for the presence of Morphine, Hydromorphone (isobaric compounds) or 6-MAM (an indicator of Heroin use). Peaks: 1. Morphine 286.1438 m/z [M+H]+ 2. Hydromorphone […]

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Nitrofurantoin Capsule Analyzed with HPLC – AppNote

Isocratic Separation of API from USP internal standard The USP assay method for the active pharmaceutical ingredient (API) found in Nitrofurantoin Capsules, uses a Phosphate Buffer and is not compatible with Mass Spectrometry. In this AppNote, using the Cogent Bidentate C18 Column, Formic Acid is used as the Mobile Phase additive, as such, the method […]

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Ocella® Tablet Analyzed with HPLC – AppNote

Separation of Drospirenone and Ethinyl Estradiol Good Selectivity with high Efficiency Peaks can be achieved analyzing Ocella Tablets. This can be helpful since Ethinyl Estradiol is present at a much lower concentration than Drospirenone and therefore can be difficult to detect; the high Efficiency can contribute to a stronger Signal in detection thus increasing Sensitivity. […]

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Oleic Acid & Elaidic Acid – AppNote

Advantages of shape selectivity in cis/trans separations The separation of cis and trans isomers by Reversed Phase can be challenging, as the two compounds often have little difference in hydrophobicity. In cases such as these, additional selectivity mechanisms may be necessary to obtain resolution. Furthermore, they cannot be resolved by MS because they have the […]

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Oleuropein in Olive Leaves Extract Analyzed with HPLC – AppNote

Separation from Matrix Peaks The main peak (Oleuropein) was observed to be well Separated from matrix components in this HPLC based approach. This Method is useful for analysts that are not using LCMS and therefore need good Chromatographic Separation in order to perform accurate Quantitation. The matrix Peaks do not co-elute with the main Peak, […]

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Oleuropein in Olive Leaves Extract Analyzed with LCMS – AppNote

High Efficiency Using a near-UHPLC Column for Oleuropein In this Application Note, the Analyte Peak is symmetrical and well Retained while the results were very reproducible (%RSD = 0.06 for Retention Times). This Method can be used to analyze and evaluate the extraction of Olive Leaves. According to the literature, Olive Leave Extracts should contain […]

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Oral OTC Anesthetic Analyzed with HPLC – AppNote

Isocratic HPLC Method for Phenol in Oral OTC Products This Method shows how an assay of Phenol can be readily performed with a simple isocratic Mobile Phase. Phenol is the active ingredient in this common Over-The-Counter oral spray solution, which is intended for treatment of sore throat. A number of inactive ingredients are present in […]

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Organic Alcohols Analyzed with HPLC – AppNote

Separation of 1-Phenyl-2-Propanol and 2-Phenyl-1,2-Propanediol In this Method, Separation is obtained between two structurally similar Organic Alcohols. The Method illustrates the type of Retention and Selectivity that can be obtained for these kinds of compounds. Furthermore, the Mobile Phase Solvents are simple to prepare and LCMS compatible. The overlay of the 5 Figures demonstrates how […]

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Orthogonal HPLC Method Development – AppNote

Choose Reversed or Normal Phase Selectivity on the Same C18 Column In the Chromatogram (A) below, a Reversed Phase Method Strategy is used: Loratadine in 100% Tetrahydrofuran (THF) elutes with the Solvent Front. With 70:30 DI Water / Tetrahydrofuran (THF), the Selectivity for this Compound is Reversed Phase, which is based on polarity of the […]

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Oxybenzone and Octinoxate Analyzed with HPLC – AppNote

Separation of Two APIs in Chap Stick® Extract This Method shows how two common ingredients found in sunscreens and lip balms can be separated. The two compounds are very hydrophobic, so a Mobile Phase Gradient with significant organic content was used in order to avoid excessive retention. Likewise, a highly organic diluent should be used […]

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Paracetamol, Aspirin and Salicylic Acid HPLC Method – AppNote

Analgesic Drugs Analyzed with HPLC A robust and rapid method was developed for the accurate simultaneous determination of Paracetamol, Aspirin, Caffeine, Benzoic Acid as an internal standard and Salicylic Acid (hydrolysis product of Aspirin). This method is rapid and sensitive and therefore suitable for routine quality control of these drugs in dosage forms and for […]

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Peonidin-3-Glucoside Analyzed by LCMS – AppNote

LC-MS method for Anthocyanin- Fruit or Vegetable Extracts In this analysis, the presence of Peonidin-3-O–Glucoside was confirmed in two out of four extracts. The peaks were symmetrical and retained beyond the dead volume. Due to the MS detection and comparison of the retention time, it was possible to find the peak for Peonidin-3-O-Glucoside among many […]

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Peptide Mixture Separated with HPLC – AppNote

Precise and Fast Equilibration Time HPLC is a technique used for the characterization of Peptides and Reversed Phase HPLC is employed for the initial analysis and final, large scale purification. The first step of the production of Synthetic Peptides usually involves an initial Separation of these compounds from its mixture, on an analytical scale, then […]

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Pepto-Bismol® Tablet Method Transfer with HPLC – AppNote

A Simple Assay Method for the widely used Pepto-Bismol® Formulation Excellent Separation of Bismuth Subsalicylate and Salicylic Acid is achieved showing that this Method can be readily transferred to a near-UHPLC 2.o phase (Figure A) from a standard 4μm Column (Figure B). The same Column Dimensions are used in both cases, with the only difference […]

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Phenolic Compound Determination with LCMS – AppNote

Compounds in Olive Leaves Extract A commercial Olive Extract was analyzed using the Cogent Phenyl Hydride Column. Only one Oleuropein Peak was detected and it was Symmetrical and well Retained. The results were reproducible (%RSD = 0.2 for Retention Times). According to the literature [1] the extract from Olive leaves should contain additional compounds. To […]

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Polycyclic Aromatic Hydrocarbons Analyzed with HPLC – AppNote

Analysis of PAH by Reversed Phase HPLC Polycyclic Aromatic Hydrocarbons (PAH) determination in soil, food, air, body fluids etc. has been a topic of interest for routine Quality Control or Screening Analyses. This Method offers good Resolution, reproducible Retention Time and Peak Shape for these compounds. In addition to the main components, small peaks due […]

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Polyphenols in Grape Seed Extract Analyzed with LCMS – AppNote

Separation of Gallic Acid, Catechin, and Epicatechin This Method was applied to the determination of Phenolics in a grape seed extract. From Figure B, it can be determined that the Aqueous extract contains a high amount of Gallic Acid, Catechin and Epicatechin. Peaks: 1. Gallic Acid 169.0142 m/z (M-H)– , RT=0.862 min 2. Catechin 289.0718 […]

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Prednisone Analyzed with HPLC – AppNote

Isocratic Method for Prednisone Tablets  This Method uses the Mobile Phase specified in the USP Method for Assay of Prednisone tablets. The resolution between the API and the Acetanilide Internal Standard meets the system suitability. Resolution was found to be superior compared to standard C18 HPLC Columns. In addition, this Method also produces excellent precision […]

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Prednisone Quantitation Method – AppNote

Simple and Easy Prednisone Analysis with LCMS Detection This Method does not require any type of derivatization. Two Gradients are presented which will allow you to choose one depending on other components in the sample and your Method objectives. Retention can be achieved on a very short column (50 mm) as shown. Linear gradient conditions […]

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Primatene Tablet Analyzed with HPLC – AppNote

Separation of Guaifenesin and Ephedrine from an OTC Formulation These two compounds are used together in the Primatene® formulation for the purpose of treating Bronchial Asthma. The Separation obtained illustrates the application of this Method in the Analysis of an Over the Counter Formulation. A wavelength change was used in order to make the two […]

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Promethazine Assay & Impurity Analyzed with HPLC – Extended AppNote

Two methods for analysis of Promethazine Tablets address difficulties associated with HPLC for these formulations. The first one discussed is the Assay Method. Here a simple isocratic approach is used and a symmetrical peak shape is obtained for the API. The method meets system suitability criteria in terms of peak area precision, tailing factor, and […]

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Pyrantel Pamoate Analyzed With HPLC – AppNote

Robust Separation with Excellent Peak Shapes The USP Assay Method for Pyrantel Pamoate uses a bare Silica Column with a Mobile Phase of Acetonitrile, Acetic Acid, Water and Diethylamine. Bare Silica Columns are often less Robust than Reversed Phase Columns due to the variable nature of the adsorbed water layer and / or the ion […]

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Quinine Sulfate and Impurity Analysis with HPLC – AppNote

API Separation from Impurity without Ion-Pair Reagents The USP Method for Quinine Sulfate requires a Resolution of not less than 1.2 from its main impurity, Dihydroquinine. In the USP Method, the ion pair agents Methanesulfonic Acid and Diethylamine are used in the Mobile Phase. Ion pair agents are often needed to reduce peak tailing of […]

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Resveratrol Capsule – AppNote

Isocratic Separation of Isomers In this easy isocratic method, the cis and trans isomers of Resveratrol are separated using a near-UHPLC stationary phase. The Cogent Bidentate C18 2.o™ Column produces excellent efficiency for both analyte peaks. The sample used here is an actual capsule formulation, demonstrating the suitability of the column for real-world samples. With […]

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Resveratrol Isomers Found with HPLC – AppNote

Resveratrol Isomers Found in OTC Capsules Two isomers of Resveratrol were observed using this simple Reversed Phase Method. Different capsules were analyzed and it was discovered that some had both isomers while the cis isomer was not detected in others (data not shown). The ability of this Method to Separate two Isomers is a good […]

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Retention of a Polypeptide Analyzed with HPLC- AppNote

Semaglutide, a Human Glucagon-like Peptide This simple isocratic Method demonstrates good Retention of a Hydrophilic Peptide where Retention is increased in Aqueous conditions.   Peak: Semaglutide Method Conditions: Column: Cogent Diamond Hydride™, 4μm, 100Å. Catalog No.: 70000-75 Dimensions: 4.6 x 75mm Mobile Phase: 60:30 DI Water with 0.1% Formic Acid / Acetonitrile. Injection vol.: 1µL […]

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Retention of a Tricyclic Amine Analyzed with ELSD- AppNote

Memantine Hydrochloride, a Tricyclic Amine As this compound lacks Chromophores, it typically requires Derivitization for use in UV detection.  In this Method, we retain excellent peak shape without the need for these pre-column derivatization steps. This compound can also cause issues with Peak Tailing in typical Reversed Phase Columns due to the strong adsorption of […]

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Rice Extract Analyzed with LCMS – AppNote

Content of Phenolic Compounds in Rice Determined by LCMS When commercial Rice Extracts were analyzed, only 6’-O-Feruloylsucrose at 3.01 minutes (Peak not shown for the clarity of the Chromatogram) was found in this particular Rice Extract Sample. Next, the Sample was spiked with the Standards according to the literature referenced below [1]. This Method is […]

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Shape and Size Selectivity of HPLC Columns – AppNote

Separation of Planar Molecules in HPLC using the Cogent UDC-Cholesterol Column When comparing Cogent UDC-Cholesterol™ and Cogent Bidentate C18™ Columns, the Cholesterol phase is generally less hydrophobic under the conditions in this Application Note. However from the two Chromatograms shown, a very interesting property of the Cholesterol Phase can be observed; the Cholesterol Column exhibits […]

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Simvastatin Analyzed with HPLC – AppNote

Excellent Peak Shape and Resolution This Method is easy to prepare, use and reproduce. Good Peak shape and Resolution is observed for Simvastatin from its hydrolysis products and tablet Excipients. Peaks: 1. Excipient 2. Hydrolysis Product 3. Simvastatin Method Conditions Column: Cogent Bidentate C18™, 4μm, 100Å Catalog No.: 40018-75P Dimensions: 4.6 x 75mm Mobile Phase: 52% DI Water / […]

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Spiked Hesperidin Extract Analyzed with LCMS – AppNote

Eriocotrin, Hesperidin, Naringin, Diosmin In this method, the Hesperidin extract was spiked with three glycosidic flavonoids. The Cogent Phenyl Hydride Column was used to separate the main component of the extract (Hesperidin) and three standards. The MS detection helped to distinguish between the Naringin peak of the standard and a small impurity peak (peak marked […]

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Stability Testing of Tetracycline HCl Capsules – AppNote

Robust, high-throughput Separation of API from its Degradation Products Figure A shows the chromatogram obtained from a single injection of the degraded Tetracycline capsule extract. Tetracycline is well-resolved from its three main degradation products, the identities of which were confirmed by individual standards under non-degrading conditions. Amine-containing analytes such as these often require the use […]

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Substituted Phenols Analyzed with LCMS – AppNote

Separation by Functional Groups Two proprietary compounds, which are precursors for a catalyst or Prodrugs are analyzed using a C18 Column under Normal Phase conditions. Separation of the two compounds is extremely reproducible (%RSD 0.1) and is very easy. The amount of moisture in your Mobile Phase is not an issue with this Method. Peaks: […]

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Sucralose – AppNote

Common Artificial Sweetener Sucralose does not exhibit significant UV absorption, hence detection of this analyte may be suited to Refractive Index (RI) as an alternative. Both approaches are shown in this application note, where UV and RI detector modules where attached in series. The relatively high concentration used in this example allowed for adequate detection […]

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Sulfisoxazole Acetyl Analyzed with HPLC – AppNote

Separation of two Common Preservatives from the API in a Pharmaceutical Suspension Orally administered suspensions with Sulfisoxazole Acetyl often contain the preservatives Methyl and Propyl Paraben. As such, it is important to separate these compounds from the API in an HPLC Assay Method to properly analyze the suspension. With this Method, excellent Separation is obtained […]

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Synthetic Peptide Analyzed with HPLC – AppNote

Peptide Production Confirmation Using the Cogent HPS© C18 An in-process Method for Qualifying a synthetic peptide is shown with 3 samples of tryptic “digests” were injected on the same Column. The Reproducibility from run to run on the samples is excellent giving the user a high level of confidence. The process and Method was further […]

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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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Tazobactam USP Assay with HPLC – AppNote

Alternate L1 Column used for the USP Assay Method of Tazobactam This USP Tazobactam Assay was easily performed and the five injection overlay below demonstrates Run to Run consistency and Precision. The Peak Efficiency is very good and Peak Tailing guidelines are easily met. This Application Note demonstrates a good Alternate Column for your Tazobactam […]

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Tetracycline USP Assay with HPLC- AppNote

USP Method for Tetracycline This USP Tetracycline Assay is performed and a five injection overlay shows run to run consistency and Precision. The Peak Efficiency is very good and Peak Tailing guidelines are easily met with RSD values less than 0.4%. This demonstrates a good Alternate Column for your USP Method.   Peak: Tetracycline Method Conditions: […]

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Tetrahydrocannabinol Analyzed with HPLC – AppNote

The Psychoactive Component THC in Cannabis  Tetrahydrocannabinol (THC) which is derived from Cannabis, has several isomeric forms, which may account for the extra Peaks observed in the Chromatogram below. Although their identities could not be confirmed, these Peaks can be Separated from the main Peak using this Method. The compound is quite hydrophobic and therefore a […]

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Ticagrelor Analyzed with HPLC – AppNote

A Reproducible Method for Detection of this Blood Clot Preventative  A rapid, sensitive, and Reproducible Method has been developed for analysis of Ticagrelor. The data below, (overlay of 5 chromatograms ) illustrates how the compound can be adequately Retained and detected using a simple Gradient in Reversed Phase HPLC.  The Method demonstrates good Peak Shape […]

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Tobramycin Analyzed with LCMS – AppNote

Retention of Highly Polar, Water Soluble Antibiotic  Tobramycin is a very polar compound that lacks a sufficient chromophore, and therefore is difficult to analyze using Reversed Phase HPLC with UV detection. In this Application Note, the Method below retains Tobramycin and Separates it Uracil which is used as a Solvent Front Marker. Using this Method, […]

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Transfer HPLC Method to UHPLC with Hydrophobic Compounds – AppNote

Separation of Hydrophobic Compounds with HPLC & UHPLC This AppNote shows separation of analytes within a range of hydrophobicity. A simple gradient is used to elute all the compounds and baseline separation is obtained for the critical pair (peaks 4 and 5) and the least hydrophobic compound is adequately retained. A comparison is shown in […]

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Urea Analyzed with HPLC- AppNote

Fast, Reproducible Method In this Method Urea was retained when a 100% DI Water Mobile Phase was used. DI Water can be used as the complete Mobile Phase without fear of phase “de-wetting” and subsequent loss of retention time. This Method has the advantage of being very simple, without time consuming derivatization or reaction steps […]

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