The intracellular life activities are jointly undertaken by many genes, proteins, and small molecule metabolites, and the functional changes of upstream macromolecules (nucleic acids, proteins, etc.) will eventually be reflected in the metabolic level, such as changes in neurotransmitters, hormones, etc. regulation, receptor effect, cell signal release, energy transfer and intercellular communication, etc., so the metabolome is in the downstream of gene regulation network and protein action network, and what it provides is the terminal information of biology. We often say that genomics and proteomics tell you what can happen, and metabolomics tells you what has happened.
What is metabolomics? What problem is metabolomics used to solve?
Metabonomics (Metabolomics) is an emerging discipline developed in the late 1990s, which is the study of metabolites (endogenous metabolites) after organisms are perturbed (such as genetic changes or environmental changes). The science of types, quantities and their changing laws. Metabolomics focuses on the study of the metabolic pathways of endogenous metabolites in the whole organism, organ or tissue, the influence of internal or external factors and the law of changes over time. Metabolomics reflects a series of biological events in a certain pathophysiological process by revealing the overall change trajectory of metabolism under the influence of intrinsic and extrinsic factors.
What are the research objects and levels of metabolomics?
In general, metabolomics focuses on small molecular compounds with molecular weights below 1000. According to different research objects and purposes, scientists divide the metabolite analysis of biological systems into four levels:
Metabolite target analysis: Qualitative and quantitative analysis of one or several specific components, such as a certain class of compounds with related structures and properties (amino acids, organic acids, cis-diols) or all intermediates or intermediates of a certain metabolic pathway. Hallmark components of multiple metabolic pathways.
Metabolite Fingerprint Analysis: Simultaneous analysis of multiple metabolites without separation and identification of specific single components.
Metabolic profiling: Rapid qualitative and semi-quantitative analysis of metabolites in specific tissues in organisms under defined conditions.
Metabolome analysis: Quantitative analysis of all metabolites contained in an organism or a specific tissue in the body, and to study the dynamic changes of the metabolome under external intervention or pathophysiological conditions.
What kinds of samples can metabolomics detect?
Metabolomics mainly studies small molecule metabolites (molecular weight <1000) that are substrates and products of various metabolic pathways. Its samples are mainly plasma or serum, urine, saliva, and cell and tissue extracts. We also cooperate with some companies or units (such as tobacco companies, tea companies, wine companies, yogurt companies, hospitals, etc.), and have done plant, fungus, microbial extracts, cerebrospinal fluid, lymph fluid, insect hemolymph, amniotic fluid, follicular fluid , Knee synovial fluid, tears, semen, placenta, feces and intestinal contents extract.
How many substances can be detected by general metabolomics?
Different mass spectrometry platforms have different sensitivity and bias, and there are complementarities between different platforms. Generally speaking, about 200 substances can be accurately characterized in serum samples by GC/MS, about 200 substances can be determined in urine samples by GC/MS, and other samples (such as liver, feces, intestinal contents) are also in the order of hundreds . If a full two-dimensional GC/MS is used, the number of substances will be more, up to more than 1,000. The number of substances detected by LC/MS is far more than that of GC/MS, but not many can be accurately qualitative and quantitative. Depending on the platform (TQ, QTOF, etc.), it can reach dozens to 2-3 hundred substances.
Sample collection method and required sample size?
– Microbial and cellular samples: rapid inactivation of metabolic activity (quenching) while keeping cells from lysis;
– Animal body fluids (such as urine, blood, tissues, organs, saliva): Pre-treatment should be performed quickly after sampling, such as adding anticoagulants, preservatives, and immediately freezing (-80 °C);
– Plant samples: frozen (liquid nitrogen) immediately after collection, and then transferred to -80℃ for storage, 200mg/case;
– Serum sample: 150uL/case, must avoid repeated freezing and thawing. Blood was collected in centrifuge tubes and allowed to stand for 30 minutes for coagulation. Centrifuge at 8000rpm for 10 minutes, take the supernatant and aliquot into cryopreservation tubes, place in liquid nitrogen or freeze at -80 degrees;
-Urine sample: 200uL/case, in principle, you can take a little more, and record the total volume of urine. For clinical samples, please take the first urine in the morning. Urine is directly dispensed into centrifuge tubes, centrifuged at 8000rpm for 10 minutes, and the supernatant is dispensed into cryopreservation tubes, placed in liquid nitrogen or frozen at -80 degrees, and frozen at -80 degrees; for other sample collection methods, please consult this article Division staff
What are the advantages of choosing metabolomics as a scientific research technique?
– Metabolomics research is in the middle of biological information flow, between genes, proteins, cells and tissues, and plays a linking role in the transmission of biological information.
– The production and metabolism of small molecules are the downstream products of gene expression, and the analysis of metabolites in biological fluids can more directly and accurately reflect the pathophysiological state of the organism.
– There are fewer types of metabolites, about 103 orders of magnitude, much smaller than the data of genes and proteins, the molecular structure of substances is much simpler, and the analysis is simpler and clearer.
– The metabolite information database of metabolomics is simple, it is far less complex than the database of whole genome sequencing and a large number of expressed sequence tags.
– Effective small changes in gene and protein expression are amplified on metabolites, making detection easier.
– Metabolites are similar in various biological systems, so the techniques used in metabolomics research are easier to use in various fields and more easily accepted.
What are the research areas of metabolomics?
-Disease mechanism research
-Disease diagnosis and prevention
-New drug screening and development
– Research on the mechanism of drug action
-Drug toxicity evaluation
– Cell metabolomic studies of plants
– Microbial metabolomics research
Why Choose Gas Chromatography-Time-of-Flight Mass Spectrometer (GC/TOF-MS)
Gas chromatography-time-of-flight mass spectrometry achieved a major technological breakthrough in the 1990s. Since time-of-flight mass spectrometry has no theoretical mass limit, the scanning speed and ion collection efficiency are very high, so it is rapidly applied to biological research.
The time-of-flight mass analyzer in the GC/TOF-MS instrument has a simple structure, has the characteristics of fast scanning and efficient ion acquisition, and has a wide mass range. /s, For the full spectrum analysis of metabolomic biological substances, the application prospect of GC/TOF-MS is very broad.
AxisPharm metabolomics analysis solutions include:
Gas chromatography-mass spectrometry (GC-MS): Commonly used for targeted analysis of water-soluble metabolites (requiring derivatization), some lipids, and organic acids.
Liquid chromatography-mass spectrometry (LC-MS): untargeted metabolomics, targeted metabolomics, untargeted lipidomics, targeted lipidomics. Commonly used for targeted and non-targeted analysis of water-soluble small molecules such as amino acids, carbohydrates, alcohols, organic acids, amines, TCA cycle intermediates, and lipid macromolecules
NMR Spectroscopy: commonly used for substance identification and analysis of simple or purified samples
*Except for easy-to-handle samples such as blood and serum, samples from cells, tissues, organs or bacteria can be subjected to metabolomic analysis.
Metabolomics Services
