PMID- 37976942
OWN - NLM
STAT- MEDLINE
DCOM- 20231216
LR  - 20240207
IS  - 1873-376X (Electronic)
IS  - 1570-0232 (Print)
IS  - 1570-0232 (Linking)
VI  - 1231
DP  - 2023 Dec 1
TI  - Analysis of D-amino acids secreted from murine islets of Langerhans using 
      Marfey's reagent and reversed phase LC-MS/MS.
PG  - 123928
LID - S1570-0232(23)00338-0 [pii]
LID - 10.1016/j.jchromb.2023.123928 [doi]
AB  - D-amino acids (D-AAs) are important signaling molecules due to their ability to 
      bind ionotropic N-methyl-D-aspartate receptors. D-serine (D-Ser), D-alanine 
      (D-Ala), and D-aspartate (D-Asp) have been found individually in the endocrine 
      portion of the pancreas, the islets of Langerhans, and/or their secretions. 
      However, there has been no report of a comprehensive assessment of D-AAs in islet 
      secretions. To evaluate the release of these compounds, the effectiveness of both 
      1-(9-fluorenyl)-ethyl chloroformate (FLEC reagent) and 
      1-fluoro-2,4-dinitrophenyl-5-L-alanine amide (Marfey's reagent, MR) in separation 
      of D/L-AA enantiomeric pairs in islet-specific buffers were evaluated. 
      MR-derivatized D/L AAs showed greater than baseline resolution (Rs >/= 1.5) of 13 
      enantiomeric pairs when using a non-linear gradient and an acidic mobile phase 
      system, while FLEC-derivatized AAs exhibited limited resolution on both biphenyl 
      and C18 columns. The optimized MR method yielded highly reproducible separations 
      with retention times less than 1% RSD. Excellent linearity between the analyte 
      concentrations and response (R(2) > 0.98) were obtained, with less than 15% RSD 
      for all analyte responses. Most analytes had an LOD at or below 100 nM, except 
      for L-Ala (200 nM). The optimized MR method was used to quantify D-AAs in 
      secretions of 150 murine islets after incubation in 3- and 20-mM glucose. In 
      response to both solutions, D-Ser and D-glutamine were tentatively identified via 
      comparison of retention time and quantifier-to-qualifer ion ratios with 
      standards, and from spiking experiments. Both were secreted in low quantities 
      which did not differ significantly in either low (D-Ser: 44 +/- 2 fmol 
      islet(-1)h(-1); D-Gln: 300 +/- 100 fmol islet(-1)h(-1)) or high (D-Ser: 23 +/- 1 fmol 
      islet(-1)h(-1); D-Gln: 120 +/- 50 fmol islet(-1)h(-1)) glucose across 3 biological 
      replicates. The method developed is robust and can be applied to further examine 
      the release of D-AAs and their potential roles in islet physiology.
CI  - Copyright (c) 2023 Elsevier B.V. All rights reserved.
FAU - Ogunkunle, Emmanuel O
AU  - Ogunkunle EO
AD  - Department of Chemistry and Biochemistry, Florida State University, 95 Chieftain 
      Way, Tallahassee, FL 32306, United States.
FAU - Davis, Joshua J
AU  - Davis JJ
AD  - Department of Chemistry and Biochemistry, Florida State University, 95 Chieftain 
      Way, Tallahassee, FL 32306, United States.
FAU - Skinner, Emily L
AU  - Skinner EL
AD  - Department of Chemistry and Biochemistry, Florida State University, 95 Chieftain 
      Way, Tallahassee, FL 32306, United States.
FAU - Thornham, James
AU  - Thornham J
AD  - Program in Molecular Biophysics, Florida State University, 95 Chieftain Way, 
      Tallahassee, FL 32306, United States.
FAU - Roper, Michael G
AU  - Roper MG
AD  - Department of Chemistry and Biochemistry, Florida State University, 95 Chieftain 
      Way, Tallahassee, FL 32306, United States; Program in Molecular Biophysics, 
      Florida State University, 95 Chieftain Way, Tallahassee, FL 32306, United States. 
      Electronic address: mroper@fsu.edu.
LA  - eng
GR  - R01 DK080714/DK/NIDDK NIH HHS/United States
PT  - Journal Article
DEP - 20231110
PL  - Netherlands
TA  - J Chromatogr B Analyt Technol Biomed Life Sci
JT  - Journal of chromatography. B, Analytical technologies in the biomedical and life 
      sciences
JID - 101139554
RN  - 0 (Amino Acids)
RN  - 95713-52-3 (Marfey's reagent)
RN  - 107474-79-3 (1-(9-fluorenyl)ethyl chloroformate)
RN  - OF5P57N2ZX (Alanine)
RN  - IY9XDZ35W2 (Glucose)
SB  - IM
MH  - Animals
MH  - Mice
MH  - *Amino Acids/analysis
MH  - Chromatography, Liquid/methods
MH  - Tandem Mass Spectrometry/methods
MH  - Alanine/chemistry
MH  - *Islets of Langerhans
MH  - Glucose
MH  - Stereoisomerism
MH  - Chromatography, High Pressure Liquid/methods
PMC - PMC10843809
MID - NIHMS1947106
OTO - NOTNLM
OT  - Chiral separation
OT  - D-amino acids
OT  - Derivatization
OT  - Islets of Langerhans
OT  - Liquid chromatography
OT  - Marfey's reagent
OT  - NMDAR
COIS- Declaration of Competing Interest The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2023/11/18 11:42
MHDA- 2023/12/17 09:42
PMCR- 2024/12/01
CRDT- 2023/11/17 18:13
PHST- 2023/07/27 00:00 [received]
PHST- 2023/10/22 00:00 [revised]
PHST- 2023/11/08 00:00 [accepted]
PHST- 2024/12/01 00:00 [pmc-release]
PHST- 2023/12/17 09:42 [medline]
PHST- 2023/11/18 11:42 [pubmed]
PHST- 2023/11/17 18:13 [entrez]
AID - S1570-0232(23)00338-0 [pii]
AID - 10.1016/j.jchromb.2023.123928 [doi]
PST - ppublish
SO  - J Chromatogr B Analyt Technol Biomed Life Sci. 2023 Dec 1;1231:123928. doi: 
      10.1016/j.jchromb.2023.123928. Epub 2023 Nov 10.