Phosphorylated Sugars Analyzed by LCMS – AppNote

Sucrose-6-phosphate, Glucose-6-phosphate, & Trehalose-6-phosphate Central Metabolic Intermediates in Plants Sugar Phosphates are very highly polar Metabolites and may be difficult to retain using traditional Reversed Phase Methods. In addition, S6P and T6P are Isobaric Compounds so it is very important to separate these two Phosphorylated Sugars for MS Detection. G6P is easily distinguished by its […]

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Phosphorylated Sugars Analyzed with LCMS – AppNote

Nucleotide Sugars UDP, ADP, TDP and CDP UDP Hexanolamine was used as the Internal Standard in the analysis of Nucleotide Sugars in this rapid analysis Method. The Mobile Phase was designed to maximize the detector response in LCMS for maximum Chromatographic Efficiency. The simple “Inverse Gradient” which produces an Aqueous Normal Phase (ANP) HPLC Method, […]

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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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Proline: Trans-4-Hydroxy-L-Proline by LCMS – AppNote

Rapid, High Efficiency Method by LCMS This method is highly specific, efficient and fast for the analysis detection, small volumes of the reaction mixture can be injected and of trans-4-hydroxy-L-proline. Due to the high specificity of the the amount of the produced compound can be determined. No derivatization is required for the detection of this […]

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Propranolol in Urine Analyzed by LCMS – AppNote

Excellent peak shape for basic compound The developed method permitted analysis of Propranolol in urine after simple sample preparation. The analysis is performed at a high concentration of Acetonitrile (Acetone may be used as well). The peak shape obtained for this basic compound was excellent. The method could be easily applied to analysis of Propranolol […]

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Purine Nucleotides Analyzed with HPLC- AppNote

Separation of Xanthosine -5’-Monophopate and Guanosine -5’-Triphosphate This Method achieves the separation of Nucleotides XMP from GTP. The analysis of these polar compounds is achieved at high concentration of an organic Solvent as part of the Mobile Phase which provides increased Sensitivity. Retention times were very Reproducible with %RSD approximately 0.4, even when Red Blood […]

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Sarcosine Analyzed with LCMS – AppNote

Separation of Potential Urine Biomarker from Isobaric ß-Alanine This developed LCMS method can separate Sarcosine from Beta- Alanine in serum and urine samples without using labor intensive sample derivatization. Since Sarcosine is considered a potential biomarker for prostate cancer risk and aggressiveness, it is essential to resolve and accurately quantify this compound in the presence […]

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Separation of Adenine Nucleotides – AppNote

Separation of AMP, ADP, ATP with HPLC The figure shows a separation of three energy nucleotides. All three are baseline separated in order of increasing polarity as is expected when ANP chromatography is used. It is worth noting that the retention of nucleotides increased as the buffer concentration was increased (data not shown). 16.0mM concentration […]

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Separation of Inosine Nucleotides – AppNote

IMP, IDP, and ITP Analyzed with HPLC The figure shows the optimized separation of ITP (Inosine 5’-monophosphate), IDP (Inosine 5’-diphosphate) and IMP (Inosine 5’-triphosphate) in the order of increasing Phosphate content similar to anion exchange. The presence of at least one impurity near ITP and possibly a second near IMP precluded accurate determination of peak […]

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Separation of Nucleotides – AppNote

AMP, NAD, NADP, and GTP on UDA Column Nucleotides are not well retained under Reversed Phase conditions due to their highly polar nature (the presence of one or more Phosphate groups). In this method, weak cation-exchange interactions can provide additional retention/selectivity along with the ANP retention of the Hydride surface. Peaks: 1. AMP – Adenosine […]

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Serotonin, Metabolites & Analogs Analyzed with HPLC with LCMS – AppNote

Simple HPLC Separation Method without Fluorescent Tags. A Method using Aqueous Normal Phase (ANP), coupled with ESI Detection (LCMS), was developed for the analysis of Neurotransmitters and their Metabolites because previous HPLC based Methods which are used to analyze NT and Metabolites in biological fluids have several drawbacks. Among other problems, they often require a […]

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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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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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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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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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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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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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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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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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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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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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