Development and validation of a sensitive LC–MS/MS method for the determination of d-serine in human plasma

• Chiral derivatization with (R)-1-Boc-2-piperidine acyl chloride.
• Quantitation of d-Ser in human plasma using standard addition method.
• Full-validation of the LC–MS/MS method in human plasma.
• Determination of circulating plasma d -Ser levels in CRPS patients.

A validated LC–MS/MS method was developed for the determination of d -Serine in human plasma. The method was fully validated for use with human plasma samples and was linear from 0.19 nmol/ml to 25 nmol/ml. The coefficient of variation was ≤5% for the high QC standards and ≤8% for the low QC standards in plasma. d -Serine and l -serine were resolved by pre-column derivatization using (R)-1-Boc-2-piperidine carbonyl chloride as the derivatizating agent. The method was used to determine the concentration of d-serine in plasma samples obtained in patients receiving a continuous 5-day intravenous infusion of (R,S)-ketamine. The changes in d-Ser levels varied in the six patients, with circulating d-Ser levels increasing as much as 35% in a patient, while decreasing 20% in a patient. While only preliminary data, the results suggests the potential importance in determining the d-Ser levels in plasma and their potential role in physiological response.

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