1,4-Benzodiazepines in Urine Analyzed with ANP – AppNote

1,4-Benzodiazepines from Urine Samples were analyzed using a Cogent Diamond Hydride 2.o HPLC Column successfully after Solid Phase Extraction (SPE). Four available Compounds (shown below) were well retained and separated. The procedure could be used for determination of this class of compounds in urine samples and other body fluids. Peaks: 1. Temazepam 301.0739 m/z [M+H]+ […]

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3-Hydroxy-3-Methylglutaric acid, HMG in Urine Analyzed with HPLC – AppNote

Important Biomarker for Diabetes and HMG Disorder is well retained with HPLC. A selective, specific, and sensitive HPLC method with LCMS Analysis has been developed for the determination of 3-hydroxy-3-methylglutaric acid (a.k.a. Meglutol) in urine samples. The method can be also used in the analysis of plasma samples after precipitation of plasma proteins with Acetonitrile. […]

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4’-Aminoacetophenone Analysis with HPLC – AppNote

4’-Aminoacetophenone Retained in Acidic Conditions This Method investigated 4’-Acetaminophenone Retention and Analysis. Adequate Retention and Sensitivity was the result and could be useful in Analysis of the Metabolites of this class of Anti-tumor Agents in body fluids (Plasma or Urine) during or after Chemotherapy. Peak: 4’-Aminoacetophenone Method Conditions Column: Cogent Bidentate C18™, 4μm, 100Å Catalog […]

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Acetyl-L-Carnitine in Plasma Analyzed with LCMS – AppNote

Spiked Plasma Samples with Acetyl-L-Carnitine The Method in this Application Note was designed to be suitable for the routine analysis of Plasma Samples obtained from animal and human Pharmacokinetic studies in which Acetyl-L-Carnitine (ALC) is administered. The calibration curve prepared for the Plasma Samples showed good Linearity (R2 = 0.999) and the Precision was good […]

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Analysis of Nucleotides – AppNote

ATP & AMP Separated in Aqueous Normal Phase (ANP) The chromatogram below shows an example of the separation possible with this Method for three Adenosine analytes using an Aqueous Normal Phase (ANP) Gradient. Peaks: 1. Adenosine-3’,5’-Cyclic Monophosphate 2. Adenosine 5’-Monophosphate (AMP) 3. Adenosine 5’-Triphosphate (ATP) Method Conditions Column: Cogent Diamond Hydride™, 4μm, 100Å Catalog No.: […]

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Analysis of Thiopurines – AppNote

Separation using LCMS Gradient Method The Cogent Diamond Hydride Column with gradient elution was used for analysis of Thiopurines. Figure A shows the Peak of 6-TG, which also can be analyzed using Isocratic conditions. Good retention and symmetrical peak shape were obtained under the analysis conditions. Figure B represents two separated Thiopurines (Mono and Di- […]

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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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Arginine, Lysine and Histidine Analyzed with LCMS – AppNote

Basic Amino Acids In Synthetic or Human Urine Can be Analyzed A “Cleanup” procedure for the isolation of the Basic Amino Acids in Urine was developed in this Method and no derivatization procedure was used. Three Basic Amino Acids were Separated using an inverse gradient or Aqueous Normal Phase (ANP) Chromatography. The advantages of this […]

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Aspartic Acid & Arginine Analyzed with LCMS- AppNote

Reduced Sample Prep and Handling Two amino acids are adequately Retained and can be easily quantified in this Method with very little Sample Prep. There is no need for any Pre-Column or Post Column derivatizing due to Mass Spectrometric Detection can detect these molecules and Aqueous Normal Phase (ANP) HPLC can Retain in their ionized […]

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ATP, UDP-Glucose UDP-Galactose Separation from Red Blood Cells – AppNote

Separation of Nucleotides  In this application, we present an easy-to-use protocol for sensitive detection and quantification of nucleotides in red blood cell extracts.  Retention times were very reproducible with %RSD around 0.4. The desired separation between UDP-Glu and UDP-Gal was achieved, all other nucleotides eluted later and did not interfere with the analysis. ATP represents […]

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Atropine Analyzed with a Gradient Method – AppNote

No Ion Pair Reagent Necessary Chromatographic separation and quantification methods of Atropa Alkaloids are often described in the literature and the method of choice is usually Ion-Pair Chromatography (IPC), which requires long equilibration times and it is not very robust. This method shows a symmetrical peak for Atropine using a simple Gradient Method that does […]

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Benzylpiperazine (BZP) – AppNote

Analysis in Hair Samples using LCMS An analytical protocol was developed for analysis of Benzylpiperazine in human hair samples. The elucidation of the chromatographic method was challenging due to the polar nature of BZP. The limits of detection / quantification for this method were determined to be 0.05 ng / mg for Benzylpiperazine in hair […]

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Cannabis Metabolite Analyzed with LCMS – AppNote

THC Metabolite Found in Urine Matrix Analyzed The extraction procedure using an SPE technique allows for successful detection of this Cannabis metabolite with this Method. It is worth noting that the Retention of this Compound was achieved using Methanol rather than Acetonitrile as the Organic component in the Mobile Phase. Peak: (l)-9-Carboxy-11-Nor-Delta-9-Tetrahydrocannabinol, m/z 345.2060 [M+H]+ […]

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Carisoprodol & Meprobamate Analysis by LCMS – AppNote

Analysis of Closely Related Compounds It has been reported that for the quantitation of Meprobamate and Carisoprodol, (in biological samples plasma, urine, hair etc) by LCMS, excessive analysis time and complex sample treatment (derivatization, solid or liquid phase extraction) may be required. The Method in this AppNote shows a rapid way to quantitate both drugs […]

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Carotenoids Analyzed with SFC – AppNote

Separation in Super Critical Fluid using a Bidentate C18 Column Cogent TYPE-C Silica™ Stationary Phases with the unique hydrophobic, Silica Hydride Surface and a very wide range of Selectivities, can be used for Supercritical Fluid Chromatography (SFC). In HPLC, Cogent TYPE-C Columns are known to retain many polar compounds in the Aqueous Normal Phase (ANP) […]

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Chenodeoxycholic Acid in Urine Analyzed with LCMS – AppNote

Chenodeoxycholic Acid (CDCA) Separation in LCMS The presence of an important bile acid, Chenodeoxycholic Acid, in human urine was detected using a simple Mobile Phase with the Method below. In addition to being simple and easy, it also provides a fast Equilibration between runs even while using a Gradient, for higher throughput. Peak: Chenodeoxycholic Acid […]

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Citric, Aconitic & Maleic Acids Analyzed with LCMS – AppNote

Low Molecular Weight Organic Acids Low mass Acids were retained and Separated in this Method with the Aqueous Normal Phase mode of HPLC. The Method was able to resolve the three compounds in 10 minutes. Limits of Detection (LOD) for them ranged from 0.05 to 24 micromolar. Peaks: 1. Maleic Acid 115.0031 m/z (M-H)- 2. […]

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Codeine and Hydrocodone in Blood Analyzed with LCMS – AppNote

LCMS Separation of isobaric Drugs Found in Urine Codeine and Hydrocodone which are Isobaric Drugs (i.e. the same m/z value) were separated using this AppNote. Multiple Samples (n=5) were prepared and analyzed. The results showed excellent Reproducibility (RSDs < 2.5%). The Method in this AppNote, after validation, can be used in Forensic Toxicological Laboratories to […]

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Corticosterones Analyzed with LCMS – AppNote

Serum Corticosterones Concentration Determinations Serum Corticosterone Concentrations are of clinical significance in Adrenal dysfunction. Its measurement is sometimes used to diagnose apparent mineral Corticoid Excess Syndrome. It can also be used as a bio marker of malignancy in Adrenal Tumors. Using this Method allows the simultaneous measurement of the two main Estrogen fractions, Estrone and Estradiol […]

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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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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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Ethanolamine Analyzed with LCMS – AppNote

Retention & Analysis of Hydrophilic, Ethanolamine from Plasma In this Application Note, the Chromatograms show Ethanolamine sufficiently retained using either the Gradient or Isocratic Methods. The advantages of using these Methods are fast equilibration between runs when Gradient analysis is required and due to the high organic content of the Mobile Phase, the Ionization efficiency […]

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Galactose-1-Phosphate Analyzed with LCMS – AppNote

Potential Clinical Screening Method for Galactosemia This Method is useful as a quantitative screening or routine clinical test to detect infants suspected of having a defect of Galactose Metabolism. It can also be used to monitor blood levels of Galactose-1-Phosphate in children with Galactosemia who are on a lactose-free diet. Peak: Galactose-1-phosphate, 259.0224 m/z (M-H)– […]

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Glutamine and Lysine Analyzed by LCMS – AppNote

Two Isobaric Amino Acid as Biomarkers Found in Saliva The Chromatogram below shows an interesting aspect of analyzing real patient samples for a pair of metabolites: Glutamine and Lysine. Figure A represents 7 samples taken from patients with cancer; Figure C shows injections of samples from 7 patients with pancreatitis while Figure B represents 7 […]

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Glutathione (GSH) Analyzed by LCMS – AppNote

LCMS Method without Derivatization The major benefit of this analysis method for Glutathione which uses the Cogent Diamond Hydride Column and MS detection is that it is performed without derivatization, unlike other LCMS approaches. A GSH signal of MS is stable after repeated analyses. The unique design of the ESI offers efficient supersoft ionization which […]

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Glyburide and Metformin Analyzed with HPLC – AppNote

Separation of “Highly Polar” and “Non-Polar” Compounds in one Isocratic run In this Method, the polar compound, Metformin, and the nonpolar compound Glyburide, can be retained on the same Stationary Phase (see A & B). Depending on the Mobile Phase composition either Metformin or Glyburide can be retained longer. Method Conditions Column: Cogent Bidentate C18™, […]

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Hesperidin Analyzed with HPLC – AppNote

LC-MS compatible method This method for analysis of Hesperidin produces an excellent analyte peak and shows separation from what appears to be two impurity peaks. The gradient method is LC-MS compatible and can be applied to many types of Hesperidin-containing samples such as citrus fruit extracts. The overlay of five runs shown in the figure […]

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Histamine and Methylhistamine Analyzed with LCMS – AppNote

No Derivatization Required for Detection or Retention with this Method Various Assay Methods for Histamine (HA) and / or its metabolite, Methylhistamine (MHA), in biological samples, have been developed. However, most of them require post-Column (for detection purposes) or pre-Column (to achieve retention) Derivatization. The Method in this Application Note provides Separation and Detection of […]

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Human Plasma Extract Analyzed by LCMS – AppNote

 Urea, Lactodifucotetraose, Trimethylamine, L-carnitine & Choline Using this Method it is possible to analyze Compounds present in a Human Plasma Samples. The mechanism of retention of these polar compounds on the Column used is unknown at this time. It is possible to Retain and Separate steroids and other Compounds based on the Shape of the […]

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Isobaric Metabolites in Urine Analyzed with LCMS – AppNote

A Simple and Reproducible LCMS Method This Method demonstrates the ability to separate a wide variety of polar metabolites using Mass Spectrometry for detection. When monitoring the EIC at 133 m/z, seven Peaks were positively detected in a Urine sample by comparison with standards or by using accurate mass. Although primarily focused on the analysis […]

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Isotretinoin in Human Plasma Analyzed with LCMS – AppNote

Isocratic, LCMS Method for Isotretinoin in Human Plasma Method development for Isotretinoin was first performed with a Reference Standard, on an HPLC with a Diode Array, UV Detector. The final, Method (Isocratic) was then transferred to an LCMS for analysis of the active ingredient in Human Plasma. The potential matrix effect on the ionization of […]

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L-a-phenylglycine Antibiotic Analysis with HPLC – AppNote

Comparison using Cogent Silica-C™ and UDC-Cholesterol™ HPLC Columns In this Method, a non-modified, silica-hydride HPLC Column, Cogent Silica-C, produces good Retention of the important amino acid L(+)-α-Phenylglycine with an Isocratic Mobile Phase. In addition the Retention Times are extremely Reproducible (%RSD 0.2). The same Method using a Cogent UDC-Cholesterol Column (UDC), silica-hydride with an 11 […]

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Leucine & iso Leucine Separated with LCMS – AppNote

Leucine and Isoleucine, Isobaric Compounds Separated for LCMS Detection This Application Note presents a Method for the Analysis of two Branch Chain Amino Acids (BCAA) with nearly identical m/z values. Addition of weak Acetic Acid to the Mobile Phase allows the small difference in the pKas of the two Isobaric Compounds to achieve Resolution. A […]

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LSD and Codeine – AppNote

LCMS Method for Spiked Urine Sample LSD and codeine may be present in Urine samples and methods are needed for their detection in these matrices. The extracted Ion Chromatograms (EICs) for both analytes are shown in the figure and illustrate excellent chromatographic as well as Mass Spectral Separation. The Method in this AppNote produces high […]

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Maleic & Fumaric Acids Analyzed with LCMS – AppNote

Isobaric Organic Acids Separated and Identified Low molecular mass, Isobaric Acids were Retained, Separated and Identified in this simple Method. The Separation of the two compounds with identical mass is necessary in order to determine which of the acids is present in the sample matrix by the Mass Spectrometer. The Method presented was found to […]

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Maleic & Fumaric Isobaric Acids Analyzed with LCMS – AppNote

Separation & Detection of Isobaric Compounds Two Isobaric Acids, Maleic and Fumaric (115 m/z) are separated when using the solvents below and a Gradient Method. Solvent A used in this application note contains 50% Methanol which is recommended when biological samples are used. This Method is reproducible and is fast to equilibrate between gradient runs […]

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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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Meprobamate Analysis by LCMS – AppNote

Rapid & Simple Analysis of Meprobamate. One Step Sample Preparation with Plasma or Urine Several methods have been reported in the literature for the quantification of Meprobamate in biological samples (plasma, urine, hair etc) for patient monitoring and other tests. Most of these methods present the drawbacks of long analysis time and complex sample pre-treatment […]

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Methionine and Glutamic Acid Analyzed with LCMS – AppNote

Non Derivatized Amino Acids From Synthetic Urine Analyzed Amino Acid analysis is generally performed with HPLC along with a time consuming Pre-Column Derivatization process that is very tedious and adds cost. Using this Method, the Derivatization step is not necessary and the compounds are easily quantified. L- Glutamic Acid can be an extremely difficult Amino […]

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Morphine Sulfate Tablet Analyzed with HPLC- AppNote

Excellent Peak Shape for Tertiary Amine Morphine can be a very problematic compound for analysis with HPLC. It has a tertiary amine and has a tendency to tail under some conditions. USP assay methodology uses phosphates, alkylsulfonates, or other LC-MS incompatible agents in the Mobile Phase. This Method uses only Formic Acid and can be […]

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Nucleobases Analyzed with HPLC – AppNote

Nucleotides Uracil, Guanine, Thymine, Adenine, Excellent Peak Shape and Resolution This Method is easy to prepare, use and reproduce. Separation is accomplished under 100% Aqueous Conditions yet there is an Alternate Selectivity. These bases may be difficult to retain on Columns with ordinary Silica that contain significant amounts of Silanols. Peaks: 1. Uracil (U) 2. […]

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Nucleotides Analyzed with LCMS – AppNote

UDP-Glucose, UDP-Galactose and Galactose-1-phosphate from Blood Cells For most physiologically relevant Nucleotides, use of Mass Spectrometry is the most practical approach for the analysis with biological samples. With Mass Spec, it is essential to Separate UDP-Glucose and UDP-Galactose chromatographically because they are Isomers or Isobaric Compounds, (same molecular weight) and cannot be easily differentiated. This […]

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Organic Acids Analyzed with LCMS – AppNote

An Important Class of Compounds Organic Acids, an important class of Metabolites, are very well Retained and Separated in this Method. Figure A shows the combined EICs for five small Organic Acids: Maleic, Aconitic, Fumaric, Citric and Oxaloacetic Acids. Two of these acids are isobaric compounds; Maleic and Fumaric Acids (115 m/z) and are separated […]

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Phenylglycine & Phenylalanine Analyzed with HPLC – AppNote

Analysis using a Simple Mobile Phase. In this Method two important amino acids: L-(+)–alpha-phenylglycine and L-phenylalanine, were Separated. C18 Columns used today which may be present in every Analytical Laboratory may not be able to Retain underivatized Amino Acids. They usually elute at or near the “void volume” with other polar compounds. Peaks: 1. L-(+)-Alpha-Phenylglycine […]

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Phospholipids Analyzed with HPLC with LCMS – AppNote

Separation of Different Classes in Porcine Brain Extract. Phospholipids are an important group of biological compounds. Their separation and using LCMS can be difficult to achieve due to MS incompatibility of reagents typically required in the Mobile Phase. In this work, Phosphatidic acids, Phosphatidylserines, and their lyso derivatives were Separated and detected in an LCMS […]

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