Aela Agela Technologies

Advance search
Quick Order
See your previous items:
You have 0 product(s) in cart

Cleanert MAS-M for the Extraction of Arachidonic Acid from faeces sample

Application Introduction

This is a LC-MS/MS method for determination of fatty acid in faeces by Cleanert MAS-M.

Favorite

Introduction

This is a LC-MS/MS method for determination of fatty acid in faeces by Cleanert MAS-M.

Experimental

Standard Material

Arachidonic acid (AA) was chose to represent fatty acid.

Stock solution was prepared by dissolved 10 mg of AA in 100mL of methanol. The stock solution was diluted with a mixture of acetonitrile﹕water (70:30, v/v) to obtain work solution.

Extraction Methods

50mg of faeces sample was added to centrifuge tube, then mixed by 0.5mL 1% formic acid in water and vortexed for 2min, added 1.5mL methanol into the tube, ultrasonic for 10min, and centrifuged under 6000r/min for 5 min. The supernatant was collected into a clean vial. Added 1.5mL methanol into the tube and repeat the extract process. Combined supernatants and filtrated by PTFE filter (0.22μm), the filtrate was collected and then mixed with 0.6mL ammonium hydroxide.

The pretreatment sample was loaded into each well of Cleanert® MAS-M plate which was conditioned with 600μL methanol and 600μL water sequentially. The plate was washed with 600 μL of water followed by 600 μL of methanol. The target compounds were eluted with 600 μL of 3% formic acid in acetonitrile. The eluates were concentrated at 45℃ under a gentle stream of nitrogen to dryness. The residues were reconstituted with 100 μL of acetonitrile﹕water (70﹕30,v/v) separately for further analysis.

Instrumentation

HPLC Column: Venusil® ASB C18 (2.1 ´ 50mm, 3μm, 150 Å)

Mobile Phase: Acetonitrile/Water = 70/30(v/v)

Flow rate: 0.2mL/min

Injection: 5µL

LC Instrument: Shimadzu LC-20A

Detection: MS (ABI4000+) – see below for conditions

Ion source: ESI-

Scan mode: MRM

Table 1 MS parameters

Analyte

IS/V

TEM/°C

GS1/ Pa

GS2/ Pa

CUR/ Pa

AA

-4500

500

55

35

15

Table 2 Precursor/Product Ions of AA

Analyte

Retention time/min

Q1

Q3

DP

CE

AA

3.3

303.0

259.1

-109

-18

303.0

230.9

-85

-29

303.0

205.1

-107

-20

303.0

177.1

-90

-30

Results and Discussion

Figure 1 Chromatogram of 5ppm AA standard


Figure 2 Chromatogram of AA in faeces sample


Figure 3 Chromatogram of faeces sample spiked with 5ppm AA standard

There was approximately 3μg/g of AA in the mouse faeces in the experiment. Background should be subtracted from the response of AA in spiked samples to calculate the recovery data.

 Table 4 Recovery data

Spiked   concentration

AA

10μg/g

88.9%

Conclusion

In this experiment we use Cleanert® MAS-M to extra fatty acid from faeces and determination by LC-MS/MS. The recovery of fatty acid is 88.9% at 10μg/g. It should be a good method to determine fatty acid.


  • Cleanert MAS (Multi-function Impurity Adsorption SPE)

    Cleanert MAS is a simplified bio-sample preparation tool which offers multifunctional adsorption capabilities to remove interferences while the analytes are remained in the aqueous phase. Although protein precipitation is the most common method in bioanalysis, it is the fact that this method is not efficient for eliminating the matrix effect on LCMS/MS due to the present of phospholipids. However w

Material Name Material Type Date
Modify product number success!
Total Price:

Close

Email To Sombody

X
Email
Subject
Content

Login

X
Email address
Password
Register Forgotten your password?