Development of a method to quantitate nematode pheromone for study of small-molecule metabolism in Caenorhabditis elegans

Kwang Youl Kim, Hyoe Jin Joo, Hye Won Kwon, Heekyeong Kim, William S. Hancock, Young Ki Paik

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Pheromones produced by Caenorhabditis elegans are considered key regulators of development, mating, and social behaviors in this organism. Here, we present a rapid mass spectrometry-based method (PheroQu) for absolute quantitation of nematode pheromones (e.g., daumone 1, 2, and 3) both in C. elegans worm bodies (as few as 20 worms) and in liquid culture medium. Pheromones were separated by ultra performance liquid chromatography and monitored by a positive electrospray ionization detector in the multiple-reaction monitoring mode. The daf-22 mutant worms were used as surrogate matrix for calibration, and stable deuterated isotope-containing pheromone was used as internal standard for measuring changes in pheromones in N2 wild-type and other strains under different growth conditions. The worm-body pheromones were extracted by acidified acetonitrile solvent, and the secreted pheromones were extracted from culture medium with solid-phase extraction cartridges. The run time was achieved in less than 2 min. The method was validated for specificity, linearity, accuracy, precision, recovery, and stability. The assay was linear over an amount range of 2-250 fmol, and the limit of quantitation was 2 fmol amounts for daumone 1, 2, and 3 in both worm bodies and culture medium. With the PheroQu method, we were able to identify the location of pheromone biosynthesis and determine the changes in different pheromone types synthesized, according to developmental stages and aging process. This method, which is simple, rapid, sensitive, and specific, will be useful for the study of small-molecule metabolism during developmental stages of C. elegans.

Original languageEnglish
Pages (from-to)2681-2688
Number of pages8
JournalAnalytical Chemistry
Volume85
Issue number5
DOIs
Publication statusPublished - 2013 Mar 5

Fingerprint

Pheromones
Metabolism
Molecules
Culture Media
Electrospray ionization
Biosynthesis
Liquid chromatography
Isotopes
Mass spectrometry
Assays
Aging of materials
Calibration
Detectors
Recovery
Monitoring
Liquids

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

Cite this

Kim, Kwang Youl ; Joo, Hyoe Jin ; Kwon, Hye Won ; Kim, Heekyeong ; Hancock, William S. ; Paik, Young Ki. / Development of a method to quantitate nematode pheromone for study of small-molecule metabolism in Caenorhabditis elegans. In: Analytical Chemistry. 2013 ; Vol. 85, No. 5. pp. 2681-2688.
@article{60465149a74d4389be2a758c34a65530,
title = "Development of a method to quantitate nematode pheromone for study of small-molecule metabolism in Caenorhabditis elegans",
abstract = "Pheromones produced by Caenorhabditis elegans are considered key regulators of development, mating, and social behaviors in this organism. Here, we present a rapid mass spectrometry-based method (PheroQu) for absolute quantitation of nematode pheromones (e.g., daumone 1, 2, and 3) both in C. elegans worm bodies (as few as 20 worms) and in liquid culture medium. Pheromones were separated by ultra performance liquid chromatography and monitored by a positive electrospray ionization detector in the multiple-reaction monitoring mode. The daf-22 mutant worms were used as surrogate matrix for calibration, and stable deuterated isotope-containing pheromone was used as internal standard for measuring changes in pheromones in N2 wild-type and other strains under different growth conditions. The worm-body pheromones were extracted by acidified acetonitrile solvent, and the secreted pheromones were extracted from culture medium with solid-phase extraction cartridges. The run time was achieved in less than 2 min. The method was validated for specificity, linearity, accuracy, precision, recovery, and stability. The assay was linear over an amount range of 2-250 fmol, and the limit of quantitation was 2 fmol amounts for daumone 1, 2, and 3 in both worm bodies and culture medium. With the PheroQu method, we were able to identify the location of pheromone biosynthesis and determine the changes in different pheromone types synthesized, according to developmental stages and aging process. This method, which is simple, rapid, sensitive, and specific, will be useful for the study of small-molecule metabolism during developmental stages of C. elegans.",
author = "Kim, {Kwang Youl} and Joo, {Hyoe Jin} and Kwon, {Hye Won} and Heekyeong Kim and Hancock, {William S.} and Paik, {Young Ki}",
year = "2013",
month = "3",
day = "5",
doi = "10.1021/ac4001964",
language = "English",
volume = "85",
pages = "2681--2688",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "5",

}

Development of a method to quantitate nematode pheromone for study of small-molecule metabolism in Caenorhabditis elegans. / Kim, Kwang Youl; Joo, Hyoe Jin; Kwon, Hye Won; Kim, Heekyeong; Hancock, William S.; Paik, Young Ki.

In: Analytical Chemistry, Vol. 85, No. 5, 05.03.2013, p. 2681-2688.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Development of a method to quantitate nematode pheromone for study of small-molecule metabolism in Caenorhabditis elegans

AU - Kim, Kwang Youl

AU - Joo, Hyoe Jin

AU - Kwon, Hye Won

AU - Kim, Heekyeong

AU - Hancock, William S.

AU - Paik, Young Ki

PY - 2013/3/5

Y1 - 2013/3/5

N2 - Pheromones produced by Caenorhabditis elegans are considered key regulators of development, mating, and social behaviors in this organism. Here, we present a rapid mass spectrometry-based method (PheroQu) for absolute quantitation of nematode pheromones (e.g., daumone 1, 2, and 3) both in C. elegans worm bodies (as few as 20 worms) and in liquid culture medium. Pheromones were separated by ultra performance liquid chromatography and monitored by a positive electrospray ionization detector in the multiple-reaction monitoring mode. The daf-22 mutant worms were used as surrogate matrix for calibration, and stable deuterated isotope-containing pheromone was used as internal standard for measuring changes in pheromones in N2 wild-type and other strains under different growth conditions. The worm-body pheromones were extracted by acidified acetonitrile solvent, and the secreted pheromones were extracted from culture medium with solid-phase extraction cartridges. The run time was achieved in less than 2 min. The method was validated for specificity, linearity, accuracy, precision, recovery, and stability. The assay was linear over an amount range of 2-250 fmol, and the limit of quantitation was 2 fmol amounts for daumone 1, 2, and 3 in both worm bodies and culture medium. With the PheroQu method, we were able to identify the location of pheromone biosynthesis and determine the changes in different pheromone types synthesized, according to developmental stages and aging process. This method, which is simple, rapid, sensitive, and specific, will be useful for the study of small-molecule metabolism during developmental stages of C. elegans.

AB - Pheromones produced by Caenorhabditis elegans are considered key regulators of development, mating, and social behaviors in this organism. Here, we present a rapid mass spectrometry-based method (PheroQu) for absolute quantitation of nematode pheromones (e.g., daumone 1, 2, and 3) both in C. elegans worm bodies (as few as 20 worms) and in liquid culture medium. Pheromones were separated by ultra performance liquid chromatography and monitored by a positive electrospray ionization detector in the multiple-reaction monitoring mode. The daf-22 mutant worms were used as surrogate matrix for calibration, and stable deuterated isotope-containing pheromone was used as internal standard for measuring changes in pheromones in N2 wild-type and other strains under different growth conditions. The worm-body pheromones were extracted by acidified acetonitrile solvent, and the secreted pheromones were extracted from culture medium with solid-phase extraction cartridges. The run time was achieved in less than 2 min. The method was validated for specificity, linearity, accuracy, precision, recovery, and stability. The assay was linear over an amount range of 2-250 fmol, and the limit of quantitation was 2 fmol amounts for daumone 1, 2, and 3 in both worm bodies and culture medium. With the PheroQu method, we were able to identify the location of pheromone biosynthesis and determine the changes in different pheromone types synthesized, according to developmental stages and aging process. This method, which is simple, rapid, sensitive, and specific, will be useful for the study of small-molecule metabolism during developmental stages of C. elegans.

UR - http://www.scopus.com/inward/record.url?scp=84874588046&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84874588046&partnerID=8YFLogxK

U2 - 10.1021/ac4001964

DO - 10.1021/ac4001964

M3 - Article

C2 - 23347231

AN - SCOPUS:84874588046

VL - 85

SP - 2681

EP - 2688

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 5

ER -