absoluteidq p180 kit biocrates life sciences ag（absoluteidq p180工具包biocrates生命科學ag)）

1、AbsoluteIDQ® p180 KitTargeted Metabolite Identification and QuantificationBringing our targeted metabolomics expertise to your lab. The Biocrates AbsoluteIDQ p180 mass spectrometry Assay Preparation Kit delivers t

2、he widest range of metabolomic information available from a single targeted assay, covering a large number of key metabolites from main metabolic pathways. The metabolome – the complete set of small molecule metabolit

3、es arising from biochemical processes in various com- partments of organisms – is dynamic and complex. The AbsoluteIDQ p180 Kit quantitatively analyzes a large number of metabolites that have already been identified as

4、 part of key biochemical pathways, providing fundamen- tal data to link changes in the metabolome to biological events.Key Features Highlights? Quantifies up to 186 metabolites in five compound classes – Acylcarnitines

5、 – Amino Acids– Biogenic Amines– Hexoses (sum of Hexoses)– Phospho- and SphingolipidsPhosphatidylcholines Lyso-Phosphatidylcholines Sphingomyelins ? Isotope-labeled and chemically homologous inter- nal standards are used

6、 for quantification– 53 analytes are fully validated as absolutely quantitative– Majority of internal standards is pre-pipetted on the Kit plate– Calibration standard mix in 7 different concentra- tions is included? Co

7、ntrols included (3 different concentration levels) ? Patented plate design ensures efficient sample derivatization and excellent reproducibility in analyte extraction? Standardized assay in 96-well plate format – suita

8、ble for manual and automated, high through- put operation? Very small sample volume of 10 µL? Allows in-house handling of valuable samples and proprietary data? Combined flow injection/liquid chromatography assay ?

9、Designed to be used with AB SCIEX or Waters triple quadrupole mass spectrometers– Industry leading triple quadrupole and hybrid quadrupole ion trap platforms provide broad linear dynamic range– Enables accurate and pre

10、cise quantification analyses– Facilitates monitoring of 100’s of MRM pairs, for wide compound coverage in a single injection? MS analysis by FIA-MS/MS method, for acylcarnitines, hexoses, phospho- and shingolipids – Ioni

11、zation via Electrospray Ionization (ESI) source– Multiple Reaction Monitoring (MRM) mode for high specificity and sensitivity– Rapid and robust assay, simple work flow– Established analysis method (following newborn scre

12、ening for inborn defects) – Employs carefully selected MRM pairs for quantification? MS analysis by LC-MS/MS method for amino acids and biogenic amines– Robust reversed-phase column – Employs carefully selected MRM pairs

13、 for quantification? 82 samples per plate? Includes MetIDQ? Software for managing the entire Kit workflow and for automated calculation of metabolite concentrations– Optional MetIDQ? StatPack Software Module for full s

14、tatistical analysis available? On-site application support for instrumentation setup, Kit preparation and software training availableAssay PreparationThe unique design of the AbsoluteIDQ Kit plate ensures quick, easy an

15、d reproducible sample preparation and efficient metabolite extraction from biological samples. By incorporating the internal standards into each well of the plate, many potential sources of error in quantifying the me

16、tabolites have already been eliminated. The patented plate is designed to perform all sample preparation steps directly on the 96-well plate. The total assay preparation time of 3-4 h includes the time required for pi

17、petting, derivatization and drying of the samples. The actual hands-on time required for preparation of a 96-well plate is only about 90 minutes.Processing the Assay in the Mass SpectrometerSample injections and LC-MS

18、analysis can be performed with all autosamplers and binary HPLC systems. A dummy plate for easy setup of the autosampler is included. MS-specific aquisition methods are provided. In the FIA-MS/MS method, each extracte

19、d sample is analyzed by a 3 min run in both positive and negative ion mode: ? 175 MRM pairs measured in positive ion mode (20 IS)? 2 MRM pairs measured in negative ion mode (1 IS)? 2 x 20 µL injection, flow rate o

20、f 30 µL/min ? Total running time per sample about 7 minIn the LC-MS/MS method, each extracted sample is chromato- graphically separated and analyzed in positive ion mode.? 69 MRM pairs measured in positive ion mode

21、(25 IS) ? 1 x 10 µL injection, flow rate of 500 µL/min LC-MS/MS-based Quantification with MS- Instrument Specific SoftwareThe calculation of metabolite concentrations after the LC-MS/MS run is performed in the

22、 mass spectrometer software? Concentrations of calibration standards and quantification methods are provided? Only a manual check of peak integration is recommended1. Add 10 µL of additional internal standard (p

23、urple), add 10 µL of plasma samples (yellow), dry 30 min under nitrogen flow2. Derivatization of amino acids and biogenic amines using 5% PITC (green), incubate 20 min, dry 45 min under nitrogen flow3. Extrac

24、t metabolites with 300 µL organic extraction solvent, shake 30 min at 450 rpm4. Filtration by centrifugation, 2 min at 100 x g.5. Remove 150 µL and transfer to second 96-deep-well plate, dilute with