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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 arachidonic acid in different samples. a: unspiked plasma of Method C, b: unspiked plasma of Method B, c: unspiked sample of Method A, d: spiked plasma of Method D at a concentration of 500 ng/mL, e: unspiked plasma of Method D, and f: arachidonic acid standard, 500 ng/mL.
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fig5: Chromatograms of arachidonic acid in different samples. a: unspiked plasma of Method C, b: unspiked plasma of Method B, c: unspiked sample of Method A, d: spiked plasma of Method D at a concentration of 500 ng/mL, e: unspiked plasma of Method D, and f: arachidonic acid standard, 500 ng/mL.

Mentions: It is noted that, as shown in Figure 5, the retention time of arachidonic acid in samples treated by Methods A to C is shifted compared with that of arachidonic acid standard. This phenomenon may be due to the accumulation of residual phospholipids and proteins on reversed phase HPLC column. In contrast, the endogenous interferences are removed efficiently in Method D by Cleanert MAS-M; the retention of arachidonic acid is stable. Also, the results listed in Table 2 reveal the advantage of Cleanert MAS-M.


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 arachidonic acid in different samples. a: unspiked plasma of Method C, b: unspiked plasma of Method B, c: unspiked sample of Method A, d: spiked plasma of Method D at a concentration of 500 ng/mL, e: unspiked plasma of Method D, and f: arachidonic acid standard, 500 ng/mL.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig5: Chromatograms of arachidonic acid in different samples. a: unspiked plasma of Method C, b: unspiked plasma of Method B, c: unspiked sample of Method A, d: spiked plasma of Method D at a concentration of 500 ng/mL, e: unspiked plasma of Method D, and f: arachidonic acid standard, 500 ng/mL.
Mentions: It is noted that, as shown in Figure 5, the retention time of arachidonic acid in samples treated by Methods A to C is shifted compared with that of arachidonic acid standard. This phenomenon may be due to the accumulation of residual phospholipids and proteins on reversed phase HPLC column. In contrast, the endogenous interferences are removed efficiently in Method D by Cleanert MAS-M; the retention of arachidonic acid is stable. Also, the results listed in Table 2 reveal the advantage of Cleanert MAS-M.

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