Monitoring phospholipids for assessment of matrix effects in a liquid chromatography-tandem mass spectrometry method for hydrocodone and pseudoephedrine in human plasma.
Matrix effects resulting in ion suppression or enhancement have been shown to be a source of variability and inaccuracy in bioanalytical mass spectrometry. Glycerophosphocholines may cause significant matrix ionization effects during quantitative LC/MS/MS analysis and are known to fragment to form characteristic ions (m/z 184) in electrospray mass spectrometry. This ion was used to monitor ion suppression effects in the determination of hydrocodone and pseudoephedrine in human plasma as a means to track and avoid these effects. The m/z 184 ion fragment was detected in both plasma extracts and solutions of phosphatidylcholine. Post-column infusion studies showed that the ion suppression for both drugs and internal standards correlated with the elution of phospholipids. HPLC conditions were adjusted to chromatographically resolve the peaks of interest from the phospholipids. Upon repeated injection, the elution time of the phospholipids decreased while elution of the analyte peaks remained unchanged. This resulted in co-elution and significantly affected peak shape and internal standard response for the analytes. It was decided to use the phospholipid fragment to monitor this matrix effect in validation samples. The resulting method demonstrated intra-day and inter-day precision within 4.5 and 5.6% for hydrocodone and pseudoephedrine, respectively, and accuracy within 8.9 and 8.7% for hydrocodone, and pseudoephedrine, respectively. There was no statistically significant difference in the internal standard response for the determination with and without monitoring the phospholipid fragment ion. We found that monitoring the phospholipid fragment was useful in method development to avoid the matrix effects, and in routine analysis to provide a practical way to ensure the avoidance of matrix effects in each individual sample.
Ismaiel
Omnia A
OA
Virginia Commonwealth University, School of Pharmacy, Department of Pharmaceutics, PO Box 980533, 410 N 12th Street, Richmond, VA 23298-0533, United States. oismaiel@vcu.edu
Halquist
Matthew S
MS
Elmamly
Magda Y
MY
Shalaby
Abdalla
A
Karnes
H Thomas
HT
D000996:Antitussive Agents / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D006853:Hydrocodone / Q000097:blood*; D010743:Phospholipids / Q000032:analysis* / Q000737:chemistry; D054199:Pseudoephedrine / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
D000996:Antitussive Agents; D010743:Phospholipids; D006853:Hydrocodone; D054199:Pseudoephedrine