Derivatization of chiral carboxylic acids with (S)-anabasine for increasing detectability and enantiomeric separation in LC/ESI-MS/MS

J Sep Sci. 2012 Nov;35(21):2840-6. doi: 10.1002/jssc.201200585. Epub 2012 Sep 24.


A simple and practical derivatization procedure for increasing the detectability and enantiomeric separation of chiral carboxylic acids in LC/ESI-MS/MS has been developed. (S)-Anabasine (ANA) was used as the derivatization reagent and rapidly reacted with carboxylic acids [3-hydroxypalmitic acid (3-OH-PA), 2-(β-carboxyethyl)-6-hydroxy-2,7,8-trimethylchroman (γ-CEHC), and etodolac] in the presence of 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholium chloride. The resulting ANA-derivatives were highly responsive in ESI-MS operating in the positive-ion mode and gave characteristic product ions during MS/MS, which enabled sensitive detection using selected reaction monitoring; the detection responses of the ANA-derivatives were increased by 20-160-fold over those of the intact carboxylic acids and the limits of detection were in the low femtomole range (1.8-11 fmol on the column). The ANA-derivatization was also effective for the enatiomeric separation of the chiral carboxylic acids; the resolution was 1.92, 1.75, and 2.03 for 3-OH-PA, γ-CHEC, and etodolac, respectively. The derivatization procedure was successfully applied to a biological sample analysis; the derivatization followed by LC/ESI-MS/MS enabled the separation and detection of trace amounts of 3-OH-PA in neonatal dried blood spot and γ-CEHC in human saliva with a simple pretreatment and small sample volume.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Anabasine / chemistry*
  • Carboxylic Acids / blood
  • Carboxylic Acids / chemistry*
  • Chromatography, Liquid / methods*
  • Dried Blood Spot Testing / methods*
  • Female
  • Humans
  • Infant
  • Male
  • Saliva / chemistry*
  • Sensitivity and Specificity
  • Stereoisomerism
  • Tandem Mass Spectrometry / methods*


  • Carboxylic Acids
  • Anabasine