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An Optimized High Throughput Clean-Up Method Using Mixed-Mode SPE Plate for the Analysis of Free Arachidonic Acid in Plasma by LC-MS/MS.

Wang W, Qin S, Li L, Chen X, Wang Q, Wei J - Int J Anal Chem (2015)

Bottom Line: The calibration curve ranged from 10 to 2500 ng/mL with sufficient linearity (r (2) = 0.9999).The recoveries were in the range of 99.38% to 103.21% with RSD less than 6%.The limit of detection is 3 ng/mL.

View Article: PubMed Central - PubMed

Affiliation: School of Material Science and Engineering, Tianjin Polytechnic University, Tianjin 300387, China.

ABSTRACT
A high throughput sample preparation method was developed utilizing mixed-mode solid phase extraction (SPE) in 96-well plate format for the determination of free arachidonic acid in plasma by LC-MS/MS. Plasma was mixed with 3% aqueous ammonia and loaded into each well of 96-well plate. After washing with water and methanol sequentially, 3% of formic acid in acetonitrile was used to elute arachidonic acid. The collected fraction was injected onto a reversed phase column at 30°C with mobile phase of acetonitrile/water (70 : 30, v/v) and detected by LC-MS/MS coupled with electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The calibration curve ranged from 10 to 2500 ng/mL with sufficient linearity (r (2) = 0.9999). The recoveries were in the range of 99.38% to 103.21% with RSD less than 6%. The limit of detection is 3 ng/mL.

No MeSH data available.


Related in: MedlinePlus

Chromatograms of proteins in samples treated with various clean-up methods, where PPT is Method A, LLE is Method B, SPE is Method C, and MAS-M is Method D.
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fig4: Chromatograms of proteins in samples treated with various clean-up methods, where PPT is Method A, LLE is Method B, SPE is Method C, and MAS-M is Method D.

Mentions: Proteins with larger molecular weight have similar retention behaviors as arachidonic acid on reversed phase HPLC column. They would cause matrix effect on the determination of arachidonic acid. As shown in Figure 4 (PPT), there are two big peaks that appeared in Method A while the rest of sample pretreatment methods are effective enough to remove proteins from plasma (Figure 4: LLE, SPE, and MAS-M). Combined with the capability for removing phospholipids and proteins, Method D with Cleanert MAS-M plate was selected for further investigation of the analysis of arachidonic acid by LC-MS/MS.


An Optimized High Throughput Clean-Up Method Using Mixed-Mode SPE Plate for the Analysis of Free Arachidonic Acid in Plasma by LC-MS/MS.

Wang W, Qin S, Li L, Chen X, Wang Q, Wei J - Int J Anal Chem (2015)

Chromatograms of proteins in samples treated with various clean-up methods, where PPT is Method A, LLE is Method B, SPE is Method C, and MAS-M is Method D.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4383463&req=5

fig4: Chromatograms of proteins in samples treated with various clean-up methods, where PPT is Method A, LLE is Method B, SPE is Method C, and MAS-M is Method D.
Mentions: Proteins with larger molecular weight have similar retention behaviors as arachidonic acid on reversed phase HPLC column. They would cause matrix effect on the determination of arachidonic acid. As shown in Figure 4 (PPT), there are two big peaks that appeared in Method A while the rest of sample pretreatment methods are effective enough to remove proteins from plasma (Figure 4: LLE, SPE, and MAS-M). Combined with the capability for removing phospholipids and proteins, Method D with Cleanert MAS-M plate was selected for further investigation of the analysis of arachidonic acid by LC-MS/MS.

Bottom Line: The calibration curve ranged from 10 to 2500 ng/mL with sufficient linearity (r (2) = 0.9999).The recoveries were in the range of 99.38% to 103.21% with RSD less than 6%.The limit of detection is 3 ng/mL.

View Article: PubMed Central - PubMed

Affiliation: School of Material Science and Engineering, Tianjin Polytechnic University, Tianjin 300387, China.

ABSTRACT
A high throughput sample preparation method was developed utilizing mixed-mode solid phase extraction (SPE) in 96-well plate format for the determination of free arachidonic acid in plasma by LC-MS/MS. Plasma was mixed with 3% aqueous ammonia and loaded into each well of 96-well plate. After washing with water and methanol sequentially, 3% of formic acid in acetonitrile was used to elute arachidonic acid. The collected fraction was injected onto a reversed phase column at 30°C with mobile phase of acetonitrile/water (70 : 30, v/v) and detected by LC-MS/MS coupled with electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The calibration curve ranged from 10 to 2500 ng/mL with sufficient linearity (r (2) = 0.9999). The recoveries were in the range of 99.38% to 103.21% with RSD less than 6%. The limit of detection is 3 ng/mL.

No MeSH data available.


Related in: MedlinePlus