Blood and Urine:
50uL – 500uL
Due to the minimal invasiveness and ease of sampling, blood (plasma, serum and whole blood) is the most commonly used sample for biomarker discovery. Metabolome data from blood samples can vary greatly with different methods of sample preparation. HMT recommends the use of EDTA-treated plasma and whole blood samples to reduce hemolysis and provide data with minimal artifacts. EDTA-treated plasma does not undergo hemolysis and will provide information about metabolites in pure plasma. In contrast, whole blood will offer information regarding metabolites in the whole blood, including blood cells.
The required volume of blood sample is 50 ~ 100µL for CE-MS analysis, and 200 – 500µL for LC-MS analysis. The samples will be suspended in your laboratory, frozen and shipped to our office.
Urine, which reflects renal function, hepatic drug pharmacokinetics, and dietary habits, is a valuable sample for metabolome analysis. Advantages of urine are that it can be sampled in a non-invasive manner and that deproteination (via ultrafiltration), followed by dilution, is sufficient preparation prior to CE-MS analysis. The required volume of urine sample is 50 ~ 100µL for CE-MS analysis. The samples will be suspended in your laboratory, frozen and shipped to our office.
30 – 100mg
The tissue samples will be weighed, suspended in our specialized tubes, and frozen in your laboratory to prevent metabolite degradation. The required volume of tissue sample is 30 ~ 50mg, and 60 ~ 100mg for CE-MS and LC-MS analysis, respectively. In HMT’s laboratory, metabolites are extracted according to established protocols selected by sample types, solvents, and other experimental conditions.
Selection of normalization methods is the starting point for the interpretation of results, but is also a critical task for metabolomics since no optimum methods that are applicable to all experimental possible paradigms. Generally, normalization of data from tissue samples is based on tissue weight. However, normalization methods based on DNA levels or protein volume may be also used. As it is not easy to determine what types of normalization is the best, HMT recommends selecting different normalization methods for different experimental systems using statistical analysis approaches, such as PCA, on a case-by-case basis.
106 -107 cells
We ask that culture cell samples be extracted and inactivated in your laboratory, to prevent metabolite degradation. The required volume of cell sample is 106 ~ 107 cells per sample, though more cells may be required in the case of exceptionally small cells (e.g. T-cells). We recommend the use of our original protocol, which is reliable, easy to perform, and requires no special reagents. However, we can also treat frozen pellets or any other type of cellular samples. Because normal saline or buffered solutions, such as phosphate buffered saline (PBS), contribute to the interference with CE-MS analysis, we will ask that a mannitol wash step be performed to reduce trace buffer salts.
Normalization of data from cultured cells is usually based on cell count. However, normalization methods based on DNA levels and protein volumes may be also used when a comparison is made among cells with different volumes per cell.
108 -109 cells
Single-cell microorganisms, such as E.coli, Yeast and Aspergillus, are used not only for basic biological research, but also in a variety of areas of study, including the production of useful materials such as proteins and fermented food. The samples will be extracted and inactivated in your laboratory to prevent metabolite degradation. The required volume of cell sample is 108 ~ 109. We recommend the use of our original protocol, which is reliable, easy to perform, and requires no special reagents. However, we can also treat frozen pellets or any other type of cellular samples.
Normalization of data from cultures cells is usually based on cell count. However, normalization methods based on DNA levels and protein volumes may be also used when a comparison is made among cells with different volumes per cell.
We will provide appropriate sampling methods, depending on your research purpose. If your samples are not listed above, please contact us.
|Blood, Body fluids||Plasma, Serum, Urine, CSF, fecal extract etc.||50 ~ 500 µL||Frozen and shipped|
|Animal tissue||Liver, Brain, Adipocytes, Muscle, Nerve, vessel etc.||30 ~ 100 mg|
|Plant tissue||Leaf, Fruit, Loot, Alga|
|Culture cells*1||Culture / Primary cell line,
Hemocyte, Sorted cells
|2 ~ 5 x 106 cells||Extracted in your Lab
(or frozen and shipped)
|Microorganism*1||Bacteria, Yeast, Fungus,
|1 ~ 10 x 108 cells
5 ~ 20 OD600 x mL
|Medium etc.||Culture medium, Beverage etc.||50 ~ 100 mL||Frozen and shipped|
|Others||Food, Soil, Insect etc.||Solids: ~ 500 mg
Liquids: ~ 1,000 µL
- *1 LC-MS analysis (Dual Scan) can not be applicable for these sample types.
- *2 Requires sample volume might be changed depending on analysis plan and study condition.
- *3 Samples will be shipped to HMT America office in Boston from United States. If you are non-US researcher, please contact us.