Development and validation of a LC-MS2 method for the simultaneous quantification of caffeine and adenosine from DBS

Rajashri Lella, Elizabeth Mary Mathew, Sudheer Moorkoth

Research output: Contribution to journalArticle

Abstract

A liquid chromatography-mass spectrometry (LC-MS) method was developed and validated for the simultaneous quantification of caffeine and adenosine in dried blood spot (DBS) samples. Caffeine is used in the treatment of apnea of prematurity in newborn children and adenosine levels play a major role in apnea. 40μL of whole blood spiked with the analytes was spotted onto specimen collection cards to prepare calibration DBS samples. 11.2 mm disks were punched from the card and was extracted in methanol and acetonitrile containing the internal standard. The extract was separated and quantified using a Phenomenex C18, 150x4.6mm, 3µm column as the stationary phase and 90 parts of 0.1% acetonitrile and 10 parts of 0.1% formic acid in water as the mobile phase delivered isocratically at 0.2mL/min. The injection volume and auto sampler temperature were 10µL and 35⁰C, respectively. The electrospray positive ion mode was used as the ionization source. The MS was operated using the single reaction monitoring at m/z 195→137.91 for caffeine and m/z 268→136.17 for adenosine. The method was seen to be linear from 0.050-10µg/ml and demonstrated 85.12-88.98% recoveries for both the analytes. The method depicted an accuracy within 99.52-100.57% and the precision (coefficient of variation) was 0.32-1.66% for both the analytes.

Original languageEnglish
Pages (from-to)5878-5882
Number of pages5
JournalResearch Journal of Pharmacy and Technology
Volume12
Issue number12
DOIs
Publication statusPublished - 12-2019

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Caffeine
Adenosine
formic acid
Apnea
Specimen Handling
Liquid Chromatography
Calibration
Methanol
Mass Spectrometry
Newborn Infant
Ions
Injections
Temperature
Water
acetonitrile
Therapeutics

All Science Journal Classification (ASJC) codes

  • Pharmacology, Toxicology and Pharmaceutics (miscellaneous)
  • Pharmacology (medical)

Cite this

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title = "Development and validation of a LC-MS2 method for the simultaneous quantification of caffeine and adenosine from DBS",
abstract = "A liquid chromatography-mass spectrometry (LC-MS) method was developed and validated for the simultaneous quantification of caffeine and adenosine in dried blood spot (DBS) samples. Caffeine is used in the treatment of apnea of prematurity in newborn children and adenosine levels play a major role in apnea. 40μL of whole blood spiked with the analytes was spotted onto specimen collection cards to prepare calibration DBS samples. 11.2 mm disks were punched from the card and was extracted in methanol and acetonitrile containing the internal standard. The extract was separated and quantified using a Phenomenex C18, 150x4.6mm, 3µm column as the stationary phase and 90 parts of 0.1{\%} acetonitrile and 10 parts of 0.1{\%} formic acid in water as the mobile phase delivered isocratically at 0.2mL/min. The injection volume and auto sampler temperature were 10µL and 35⁰C, respectively. The electrospray positive ion mode was used as the ionization source. The MS was operated using the single reaction monitoring at m/z 195→137.91 for caffeine and m/z 268→136.17 for adenosine. The method was seen to be linear from 0.050-10µg/ml and demonstrated 85.12-88.98{\%} recoveries for both the analytes. The method depicted an accuracy within 99.52-100.57{\%} and the precision (coefficient of variation) was 0.32-1.66{\%} for both the analytes.",
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Development and validation of a LC-MS2 method for the simultaneous quantification of caffeine and adenosine from DBS. / Lella, Rajashri; Mathew, Elizabeth Mary; Moorkoth, Sudheer.

In: Research Journal of Pharmacy and Technology, Vol. 12, No. 12, 12.2019, p. 5878-5882.

Research output: Contribution to journalArticle

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