10 Phenolic Acids Analyzed with LCMS – AppNote

Compounds Shown were Extracted from Commercial Rice. Commercial Rice Extract 3 was spiked with ten standards and analyzed. The Cogent Phenyl Hydride Column was an excellent choice to use for analysis of Phenolic Compounds with Reversed Phase HPLC. The Retention and Separation of all nine available standards was possible using the Method below.   Peaks: […]

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5 Phenolic Compounds Separated by LCMS – AppNote

Commercial Rice Extract A commercial rice extract was analyzed and peaks were assigned based on retention times and m/z values for the compounds of interest using the Cogent Phenyl Hydride Column . After method validation the developed procedure can be used to evaluate the quality of rice and to develop the best extraction procedure. Peaks: […]

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Aegeline in Protein Sport Mix – AppNote

LC-MS analysis of harmful substance in food supplement The data demonstrates the use of the Cogent Bidentate C18 2.o™ Column for analysis of dietary supplement samples. Isolation of the Aegeline peak is readily obtained with an extracted ion chromatogram (EIC) in the positive ion mode. Chromatographically, the peak is well retained and highly symmetrical. The […]

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Analysis of Alpha and Beta-Acids in Hops with HPLC – AppNote

Robust, Precise Method for Beer Analysis with HPLC The effectiveness of this Method is evident from the fact that data obtained were in a good agreement with the literature values. This Method also has the added bonus of good Precision (%RSD =0.1 and below). Calibration Curves for Alpha and Beta Acids in Beer showed good […]

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Analysis of Harmful Substances in Dietary Supplements – Extended AppNote

Dietary Supplements have had, in notable cases, ingredients added to them that have been implicated in serious health issues and even death. Two such pernicious compounds are Aegeline and 1,3-Dimethylamylamine. These two can be readily separated from the dietary supplement matrix using the Cogent Bidentate C18 2.o™   and Cogent Diamond Hydride™ Columns by LCMS […]

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Analysis of Litchi Seed Extracts – Extended AppNote

Hypoglycin A and Methylenecyclopropylglycine are two compounds found in the extract of Litchi Seeds that studies have shown to be toxic. Methylenecyclopropylglycine inhibits the synthesis of fatty acids from Glucose, and therefore undernourished children are more at risk since their Glucose reserves are low. This study used LCMS with a Cogent Diamond Hydride™ Column to […]

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Anatoxin-a, ANTX-A Analyzed with LCMS – AppNote

Potent Neurotoxin Analyzed without Derivatization Anatoxin-a or ANTX-A is a low molecular weight compound with a secondary amine which can be difficult to retain with most Reversed Phase HPLC Columns. This Application Note illustrates a very simple Method for Retention of Anatoxin-a which does not require pre-Column or post Column Derivatization as with other Methods. […]

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B Vitamins Analysis with HPLC – AppNote

Separation of Five Highly Polar Vitamins The B Vitamins are all highly polar and often ion pair agents are needed in Reverse Phase in order to retain them. However, these agents are incompatible with LCMS and therefore limit the applications of these methods. In this method using the Diamond Hydride Column, good separation is obtained […]

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Bisphenol A, BPA Analyzed with HPLC – AppNote

Toxic Substance Found in Consumer Products Analyzed with HPLC Bisphenol A (BPA) is a challenging compound for analysis with HPLC. Monitoring of this environmental and health toxicant is a necessary process for surveillance as well as risk assessment. As shown in the 5 Chromatograms overlaid below, this Method produces Peak Shapes that are Symmetrical and […]

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Bisphenol A, S, & F Analyzed with HPLC – AppNote

Separation of Epoxy Resins Found in Consumer Products & Food Many consumer products are using Bisphenol A (BPA) alternatives in recent years due to health concerns regarding toxicity of these compounds.  An analytical HPLC method is shown in this AppNote to distinguish amongst these various Bisphenol compounds potentially in chemicals and foods. Excellent resolution was […]

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Brazilian Tree Bark Analyzed with LCMS – AppNote

Identification of Quercetin, Rutin, and “Compound 1” in a Brazilian Tree Bark Over one quarter of natural medicines have been discovered in rainforests, and therefore analytical study of the indigenous flora and fauna is necessary. The Amazonian rainforest in Brazil is the most biodiverse region in the world. A Brazilian tree bark extract (Brownea Grandiceps) […]

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Caffeine Content in Coffee Analyzed with HPLC – AppNote

Espresso, Regular, and De-Caffeinated Coffee Caffeine Content Caffeine is separated from other matrix components in three common Coffee beverages in this AppNote. A shot of Espresso (Figure A) had the highest content, followed by regular drip Coffee (Figure B) as expected. In De-Caffeinated Coffee, Caffeine is extracted from the coffee beans but even after numerous […]

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Caffeine in Coffee Analyzed with HPLC- AppNote

Unique Retention Mode Affords Superior Specificity Although Caffeine retains well in Reversed Phase, it is found to be difficult to obtain a well-resolved Peak free of interference from other matrix peaks with a complex sample such as coffee. In this Method, most of the matrix peaks elute near the void volume and do not interfere […]

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Capsaicin in Topical Solution Analyzed with HPLC – AppNote

Separation of Capsaicinoids in an OTC Formulation A  formulation of Capsaicins is used as a topically applied solution for Arthritis pain relief. Analyzing Capsaicin and a similar compound, Dihydrocapsaicin is required. The Method illustrates the ability to resolve structurally similar Capsaicinoids from a pharmaceutical formulation. Peak Shapes and Efficiencies are excellent for both Peaks. Furthermore, […]

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Carotenoids Analyzed with HPLC – AppNote

Compounds of the Carotenoid Family Retained These compounds are very lipophilic and often have subtle differences in structure. Resolution of these compounds can be difficult with a standard Reversed Phase C8 or C18 Column because of these Similarities. The Cogent C30 Column on the other hand can further differentiate by analyte Shape in addition to […]

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Catecholamines Analyzed with LCMS – AppNote

Separation of Norepinephrine, Epinephrine, and Dopamine hydrochloride This Method shows separation of similar Catecholamines analyzed with Atmospheric Pressure Chemical Ionization (APCI) in Positive mode MS due to the lack of suitable chromophores on these molecules.  Proper separation of these compounds is important in assessment of biological samples. Peaks: 1. L-alpha– methyl – dopa  2. Norepinephrine […]

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Cyanidin-3-O-Glucoside Analyzed in Fruit – AppNote

Analysis in Fruit and Vegetable Extracts In this analysis of proprietary fruit & vegetable extracts, the presence of Cyanidin-3-O-Glucoside was confirmed in three out of four extracts. The peaks were Symmetrical and Retained beyond the dead volume. The Method after validation can be used to quality control of commercial fruit extracts as well in studies […]

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Cyanuric Acid and Melamine Analyzed with LCMS – AppNote

Simple Gradient Method with MS Detection This Method by LCMS detection eliminates derivatization which may be necessary with GCMS Methods.  Reversed Phased Methods may run into poor separation of the small and very polar Melamine, resorting to ion pairing reagents to improve separation.  The use of these ion pairing reagents counterintuitively results in ionization suppression […]

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Domoic Acid Analyzed by LCMS – AppNote

Determination without Derivatization To date, several analytical methods have been developed for the quantitative determination of Domoic Acid (DA). However, many of the methods require tedious and labor intensive derivatization of this compound in order to detect it. A new method based on HPLC and MS detection which does not require derivatization has been developed. […]

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Domoic Acid in Seafood Samples Analyzed by LCMS – AppNote

Domoic Acid (DA) From Seafood Without Derivatization Several Seafood and Plankton samples were tested for Domoic Acid (DA) which, as a highly polar compound, is poorly Retained on most Reversed Phase Columns. This developed Method based on HPLC and MS detection does not require derivatization of Domoic Acid. The concentration of this potent toxin varied […]

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Effect of Sodium Acetate for Glucose Ionization in LCMS – AppNote

Improved Signal to Noise for Glucose with LCMS Glucose, a simple monosaccharide, was analyzed by LCMS and the peak is very symmetrical and easy to integrate. This application note illustrates the importance in the addition of Micro Molar amount of Sodium to the Mobile Phase when sugars are analyzed. Sodium adducts of sugars produce much […]

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Efficiency of ANP Measured – AppNote

Aqueous Normal Phase (ANP) Efficiency Measured with Sorbitol, Glucose & Uracil Figure A is an excerpt from a study that investigated analyses of Glucose and Sorbitol on TYPE-C Silica based HPLC Columns and found that the efficiency of ANP is better than observed with HILIC. A Cogent Diamond Hydride Column (2.1 x 150mm with a […]

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Epigallocatechin Gallate EGCG Assay with HPLC – AppNote

A Reliable Assay for Detection of a Catechin found in Green Teas   This simple Isocratic Method may be used to quantitate this potentially therapeutic compound found in plants.  The Method maintains consistent and reliable results. The %RSD for Retention Times for both standards and sample were less than 0.1%. Peak: EGCG Method Conditions: Column: […]

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Folic Acid Analyzed by LCMS – AppNote

Minimized Sample Prep & Increased Sensitivity for Vitamin B9 A sensitive and reliable Method was developed for the determination of Folic Acid (FA) and can be used in many applications including complex sample matrices without the Solid-Phase Extraction clean up. Since the method uses LC-MS (negative-ion) it provides sensitivity as well as great selectivity. The […]

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Folic Acid Content in Fortified Cereals Analysis with HPLC – AppNote

Online Cleanup of Sample or Matrix Components This application note demonstrates an effective means of separating and resolving Folic Acid from undesired matrix components in cereal extracts without the use of SPE or other sample cleanup techniques. SPE is normally performed to remove compounds from the matrix which would co-elute with Folic Acid before injection […]

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Fructose and Glucose in Coca-Cola Analyzed with HPLC & RI – AppNote

Separation of Polar Sugars in Coca Cola© The high sugar content of soda beverages can be observed in this Chromatographic separation of Fructose and Glucose in cola. The two Sugars are retained and separated with the use of the Cogent Amide Column. Data from reference standards is also shown for peak identity confirmation. A preconditioning […]

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Galactose and Sucrose – AppNote

Retention and Separation of Mono and Disaccharides This application demonstrates the ability of the Cogent Amide Column to retain two highly polar test solutes. Sucrose, for example, has a log P of -3.7 and hence would be unlikely to retain in Reversed Phase. Excellent separation is obtained here for these Mono and Disaccharides. The noisy […]

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Glycosidic Analyzed with LCMS – AppNote

Cyanidine-3-Hexoside, Luteolin 7-O-Glucoside & Malvidin Dihexoside Observed in Mesquite Flour extracts, these compounds are Glycosidic Flavones and Anthocyanins. The Glycoside Moieties impart polar characteristics to the analytes, which makes them amenable to Retention by the Aqueous Normal Phase (ANP). In this Application Note, the Cogent Phenyl Hydride Column was used to obtain Chromatographic Separation. The […]

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Glycosidic Flavonoids Analyzed with HPLC – AppNote

Separation of Diosmin, Eriocitrin, and Naringin In this method, three Glycosidic flavonoid standards are separated with good resolution. The Cogent Bidentate C18 2.o™ Column produces high efficiency peaks with reproducible retention (see figure overlay). Separation is also observed for a small impurity peak, eluting between peaks 1 and 2. This separation of standards could be […]

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Histamine Analysis in Tuna by LCMS – AppNote

LC-MS method without derivatization A small amount of Histamine was found in a Tuna sample (Figure A) after an extraction procedure and analysis using the Cogent Diamond Hydride Column and MS detection. In Figure B, a Tuna sample was spiked before the extraction procedure at a level of 0.5 mg/L and Figure C shows the […]

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Histamine Analyzed with HPLC – AppNote

LCMS Compatible Method for the Separation of Histamine As a Heterocyclic primary amine, Histamine is a very polar compound and is difficult to retain on a Reversed Phase Column. This Method provides adequate Retention and a symmetrical Peak shape for this challenging compound without the use of ion pairing reagents in the Mobile Phase. The […]

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Histamine Content in Red Wine Using IPA – AppNote

Use Isopropyl Alcohol for Improved Peak Shape of Histamine  This Method for Red Wine Analysis is simple, fast, robust and does not require derivatization like many other HPLC Methods.  When Isopropanol (IPA) was not used in the A Solvent, the Histamine Peak was highly Symmetrical in the standard (Compare Figure C and D) but distorted […]

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Histamine in Cheese Analyzed by LCMS – AppNote

No Derivatization Required for this Analysis of Histamine A small but measurable amount of Histamine was found in a Cheese sample (Figure A) after an extraction procedure and analysis using the Cogent Diamond Hydride Column with MS detection. Two spiked Cheese samples were also analyzed. In Figure B, the Cheese sample was spiked before the […]

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Histamine in Red Wine Analyzed with LCMS – AppNote

Demonstrating Low Histamine Content in Red Wine Samples Two Red Wine samples were tested for Histamine using the Cogent Diamond Hydride Column in an LCMS method. The wine shown in Figure B was claimed to have virtually no Histamine but had no analytical evidence to support it. The claim was verified using this simple LCMS […]

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Hydrophilic Vitamins Analyzed with LCMS – AppNote

Different Mobile Phase for LCMS from a UV Method was Needed Hydrophilic Vitamin analyses were developed using UV detection. However, when the same Method and Mobile Phase was tried with MS Detection, the Peak intensities were very low and Peak Shape for Pyridoxine was not acceptable (see Red Peaks in Figures below). The Mobile Phase […]

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L-Theanine in Green Tea Analyzed with HPLC – Extended AppNote

L-Theanine in Green Tea L-Theanine does not have significant UV absorption, but detection at a low wavelength (200 nm) was found to be suitable. With an expected sample concentration of around 10ppm, a calibration curve in the range 2–200 ppm would be appropriate. The data showed good linearity in the studied range, with a line […]

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Limonin in Orange Juice Analyzed by LC-MS – AppNote

Separation from Matrix Peaks with Reproducible Results Analysis of Limonin in Spiked Orange Juice was performed using a simple isocratic method with MS detection. Analysis at 471.2 m/z allowed for detection of only the Limonin peak without interference of the juice matrix. When the Filtered, Unspiked Orange Juice was injected, a small peak for Limonin […]

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Limonin in Orange Juice Analyzed with HPLC – AppNote

Reproducible Separation from Sample Matrix Peaks An Orange Juice Sample was spiked at the level of 250 ppm of Limonin and filtered (MicroSolv Tech Corp. 0.45μm Nylon Syringe Filter)  before injection. Several Peaks from to the Juice Sample Matrix were separated (peaks1–5), the Limonin peak was well separated from the other Peaks and was easy […]

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Limonin Repeatability in HPLC Analyses – AppNote

Separating Limonin from Matrix Peaks When Limonin determination is done with HPLC, the main drawback of some Methods is poor Reproducibility of the results. Using the Method in this AppNote, a simple Isocratic Method, helped to achieve acceptable Repeatability of the results. The %RSD for Peak Areas was 1.64 for 10 consecutive injections. Peak 1 […]

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Melamine Analyzed with LCMS – AppNote

Detect Trace Levels of Melamine The unique chemical properties of Melamine (base functionalities) present challenges in obtaining Retention and adequate Sensitivity, when using standard HPLC Columns. In this Method, Melamine is Analyzed under Aqueous Normal Phase (ANP) conditions with an increased Sensitivity making it possible for detection at Trace Levels. Peak: Melamine m/z 127 (M+H)+ […]

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Milk Extract Analyzed by LCMS – AppNote

Analysis of Composition using LCMS Milk components are polar compounds that are not well retained or resolved by traditional Reversed Phase Chromatography. In addition, many of the compounds do not contain strong chromophores, resulting in low specificity and sensitivity in UV detection. The analysis using the Cogent Diamond Hydride Column and MS detection provides separation […]

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Mycotoxins Analyzed with LCMS – Extended AppNote

Ochratoxin A, Zearalenone, Patulin, Aflatoxin B1, B2, G1+G, Fumonisin B1 The detection of Mycotoxins has been an area of increasing focus in ensuring human health due to their high toxicity even at low concentrations. These chemicals are produced by various species of mold, including Aspergillus Penicillium, Paecilomyces, and Fusarium types. Exposure can come from ingestion […]

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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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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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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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