October 16, 2018
Micro Volume QuEChERS Kit for LC/MS (Forensic)
Biological Sample Pretreatment Kit
This pretreatment kit for human biological samples is intended for the field of forensic medicine. Salt precipitation via QuEChERS extraction salts is adopted as the method* for extracting toxicants in blood and urine. The sample volume has been scaled down to 100 μL, so biological samples can be pretreated even if an ample sample volume cannot be ensured. Samples pretreated with this kit can be measured with high sensitivity using LC/MS/MS.
* A patent application has been submitted for this technology.
Ready-to-Use Pretreatment Kit
A 2.0 mL tube in the Micro Volume QuEChERS Kit contains the QuEChERS extraction salts (100 mg), which eliminates the need to weigh out the QuEChERS extraction salts. As a result, if an urgent analysis is required, samples can be pretreated quickly by following the workflow shown below. Additionally, this eliminates variance in QuEChERS extraction salt volumes due to manual processing, resulting in data with high reproducibility.
Pretreatment Protocol
The Micro Volume QuEChERS kit is intended for the use in pretreatment to extract a medicinal toxicant from a biological material. The composition of QuEChERS extraction salt complies with AOAC2007.01.
This instruction manual presents an example of pretreatment following the protocol of “LC/MS/MS Method Package Rapid Toxicology Screening Ver.2“. The following shows the pretreatment flow. For the details, see “4. Pretreatment Method (Example)“.
1. Add a sample
Add acetonitrile of 300 μL and distilled water of 200 μL to the QuEChERS extraction salt.
2. Mix the solution
Mix them homogeneously with the vortex mixer.*
Dilute the 100 ng/μL Diazepam-D5 solution, which is a specified item, with acetonitrile to 10 ng/μL.
Add the 10 ng/μL Diazepam-D5 solution of 2 μL and the 1000 ng/μL Phenobarbital-D5 solution of 1 μL.
Add a sample of 100 μL such as whole blood, blood serum, or urine.
With the vortex mixer, mix the resultant solution sufficiently so that the QuEChERS extraction salt dissolves completely.
* The QuEChERS extraction salt may absorb moisture to form lumps. Mix it sufficiently to homogeneity in the lumps.
3. Centrifuge the solution
As soon as finishing mixing the solution, centrifuge the solution at room temperature (15000 g, 10 minutes).
4. Transfer the supernatant, Add the 0.1 % TFA solution
Transfer the supernatant to a new 1.5 mL tube or 2.0 mL tube.*
Add the 0.1 % TFA acetonitrile solution of 20 μL and mix the resultant solution with the vortex mixer.
* Using the LCMS-8060 for analysis allows you to analyze the supernatant as it is.
5. Dry and concentrate the solution with nitrogen gas
With the drying and concentrating unit capable of spraying nitrogen gas.
6. Add methanol as a new solvent
Add methanol of 40 μL to the dried sample and then dissolve it with the vortex mixer.
7. Centrifuge the solution
Centrifuge the resultant solution at room temperature (15000 g, 5 minutes).
8. Dispense the supernatant to the vial
Dispense the supernatant to the vial for HPLC.
Set the vial in the autosampler to analyze it. For analysis conditions, refer to the instruction manual of “LC/MS/MS Method Package Rapid Toxicology Screening Ver.2”.
Example of Use: Toxicant Additive Recovery Test
Using the Micro Volume QuEChERS Kit, an additive recovery test was implemented for the main toxicants in whole human blood. The measurements were performed by LC/MS/MS using the HPLC and MS conditions registered in the LC/MS/MS Rapid Toxicology Screening System Ver.2. Favorable results were obtained in this experiment, with recovery rates* for all compounds ranging from 71% to 86%. Using the Micro Volume QuEChERS Kit, impurities can easily be removed, enabling the efficient recovery of a wide range of compounds without loss.
* The recovery rate is calculated using the following formula.
Recovery rate = (Area for the pre-added compound) / (Area for the post-added compound) x 100
Toxicant Additive Recovery Rate
These data were obtained from Associate Professor Kei Zaitsu and Kengo Matsumoto, Department of Legal Medicine and Bioethics, Nagoya University.
Product Specifications
| Item | Specifications |
|---|---|
| Applicable Samples | Whole blood, urine and other biological samples |
| Use Cycles | Once/pc. |
| Sample Volume | 100 μL |
| Quantity | 100 pc. |
Product Configuration (per pc.)
| Item | Volume | Remarks |
|---|---|---|
| QuEChERS Extraction Salt | 100 mg | As per AOAC 2007.01 Magnesium sulfate: 80 mg Sodium acetate: 20 mg |
Note) When storing this product, avoid direct sunlight as well as high temperatures and high humidity.
For the analysis conditions, refer to the LC/MS/MS Rapid Toxicology Screening System Ver.2 and the LC/MS/MS Forensic Toxicological Database.
(455 KB)- This product is for research purposes only. It cannot be used for clinical diagnostic applications.
- Shimadzu assumes no responsibility for any damages incurred directly or indirectly from the use of this product. The customer assumes responsibility for the results and phenomena arising from its use. Additionally, note that the usefulness of information obtained from using this product cannot be guaranteed.