Summary.
A novel sample preparation protocol for gas chromatographic (GC) analysis of amino acid enantiomers in peptides was developed. It comprises traditional acid hydrolysis, a novel treatment of the analytes with a fluoroalkyl chloroformate and GC/FID separation of enantiomers on a chiral capillary column. The major improvements consist in that the derivatization step proceeds in organic-aqueous media within seconds and the amino acid derivatives are volatile enough to suit the temperature range of the chiral Chirasil-Val® capillary column. The approach was found beneficial for chiral analysis of pharmaceutically important Carbetocin peptide.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
CM Adams RA Zubarev (2005) ArticleTitleDistinguishing and quantifying peptides and proteins containing D-amino acids by tandem mass spectrometry Anal Chem 77 IssueID14 4571–4580 Occurrence Handle16013875 Occurrence Handle10.1021/ac0503963 Occurrence Handle1:CAS:528:DC%2BD2MXltlGgu7c%3D
EJ Ariëns (1986) ArticleTitleStereochemistry: A source of problems in medicinal chemistry Med Res Rev 6 IssueID4 451–466 Occurrence Handle3534485 Occurrence Handle10.1002/med.2610060404
T Barth J Slaninová L Servítová M Lebl Ch Kotsira-Engopoulos Ch Tzougraki I Photaki M Krojidlo K Jošt (1981) ArticleTitleThe effect of some structural modifications of the oxytocin molecule on the duration of the galactogogic response Coll Czech Chem Commun 46 2441–2445 Occurrence Handle1:CAS:528:DyaL38XpslygsA%3D%3D
E Bayer G Nicholson H Frank (1987) Separation of amino acid enantiomers using chiral polysiloxanes: quantitative analysis by enantiomer labelling ChW Gehrke KCT Kuo RW Zumwalt (Eds) Amino acid analysis by gas chromatography NumberInSeriesII CRC Press Boca Raton 46–48
H Brückner M Wachsmann (2003) ArticleTitleEvaluation of fluorescent monochloro-s-triazine reagent for amino acid analysis Chromatographia 57 IssueIDSuppl. S S143–S146 Occurrence Handle10.1007/BF02492096
H Brückner T Westhauser H Godel (1995) ArticleTitleLiquid-chromatographic determination of D-amino-acids and L-amino-acids by derivatization with o-phthaldialdehyde and N-isobutyryl-L-cysteine – applications with reference to the analysis of peptidic antibiotics, toxins, drugs and pharmaceutically used amino acids J Chromatogr A 711 IssueID1 201–215 Occurrence Handle7496491 Occurrence Handle10.1016/0021-9673(95)00158-J
BJ Clark JE Mama (1989) ArticleTitleResolution of chiral compounds by HPLC using mobile phase additives and a porous graphitic carbon stationary phase J Pharmaceut Biomed Anal 7 IssueID12 1883–1888 Occurrence Handle10.1016/0731-7085(89)80208-0 Occurrence Handle1:CAS:528:DyaK3cXltVemsLY%3D
N Cort S Einarsson G Astrom (1982) ArticleTitleEffect of oxytocin and its long-acting analog on milk let-down and intra-mammary pressure in healthy lactating sows Am J Vet Res 43 IssueID7 1283–1285 Occurrence Handle7103212 Occurrence Handle1:CAS:528:DyaL38Xks1Kks78%3D
H Eckert B Forster (1987) ArticleTitleTriphosgene, a crystalline phosgene substitute Angew Chem Int Ed Engl 26 894–895 Occurrence Handle10.1002/anie.198708941
H Frank (1987) Analysis of amino acid racemization and the separation of amino acid enantiomers ChW Gehrke KCT Kuo RW Zumwalt (Eds) Amino acid analysis by gas chromatography NumberInSeriesII CRC Press Boca Raton 56–66
X Fu T Do FJ Schmitz V Andrusevich MH Engel (1998) ArticleTitleNew cyclic peptides from the ascidian Lissoclinum patella J Nat Prod 61 IssueID12 1547–1551 Occurrence Handle9868162 Occurrence Handle10.1021/np9802872 Occurrence Handle1:CAS:528:DyaK1cXntVWltrw%3D
ChW Gehrke LL Wall JS Absheer FE Kaiser RW Zumwalt (1987) Sample preparation for chromatography of amino acids: acid hydrolysis of proteins RW Zumwalt KCT Kuo ChW Gehrke (Eds) Amino acid analysis by gas chromatography NumberInSeriesI CRC Press Boca Raton 2–5
DR Goodlett PA Abuaf PA Savage KA Kowalski TK Mukherjee JW Tolan N Corkum G Goldstein JB Crowther (1995) ArticleTitlePeptide chiral purity determination – hydrolysis in deuterated acid, derivatization with Marfey reagent and analysis using high-performance liquid-chromatography electrospray-ionization mass-spectrometry J Chromatogr A 707 IssueID2 233–244 Occurrence Handle7633594 Occurrence Handle10.1016/0021-9673(95)00352-N Occurrence Handle1:CAS:528:DyaK2MXntl2ns7g%3D
G Gübitz MG Schmid (2006) ArticleTitleChiral separation principles in chromatographic and electro migration techniques Mol Biol 32 IssueID2 159–179
S Hess KR Gustafson DJ Milanowski E Alvira MA Lipton LK Pannell (2004) ArticleTitleChirality determination of unusual amino acids using precolumn derivatization and liquid chromatography-electrospray ionization mass spectrometry J Chromatogr A 1035 IssueID2 211–219 Occurrence Handle15124814 Occurrence Handle10.1016/j.chroma.2004.02.068 Occurrence Handle1:CAS:528:DC%2BD2cXivVGjtL0%3D
P Hušek (2005) Quantitation of amino acids as chloroformates – a return to gas chromatography I Molnár-Perl (Eds) Quantitation of amino acids and amines by chromatography Elsevier Amsterdam 2–38
Hušek P, Šimek P, Hartvich P, Zahradníčková H (2007) Fluoroalkyl chloroformates in treating amino acids for gas chromatographic analysis. J Chromatogr A (in press)
KE Kawulka T Sprules CM Diaper RM Whittal RT McKay P Mercier P Zuber JC Vederas (2004) ArticleTitleStructure of subtilosin A, a cyclic antimicrobial peptide from Bacillus subtilis with unusual sulphur to alpha-carbon cross-links: Formation and reduction of alpha-thio-alpha-amino acid derivatives Biochemistry 43 IssueID12 3385–3395 Occurrence Handle15035610 Occurrence Handle10.1021/bi0359527 Occurrence Handle1:CAS:528:DC%2BD2cXhs1yit78%3D
JW LeFevre EJ Gublo C Botting R Wall A Nigro MLT Pham G Ganci (2000) ArticleTitleQualitative reversed-phase thin-layer chromatographic analysis of the stereochemistry of D- and L-alpha-amino acids in small peptides J Planar Chromat 13 IssueID3 160–165 Occurrence Handle1:CAS:528:DC%2BD3cXmvFKhtb0%3D
YM Liu M Schneider CM Sticha T Toyooka JV Sweedler (1998) ArticleTitleSeparation of amino acid and peptide stereoisomers by non-ionic micelle-mediated capillary electrophoresis after chiral derivatization J Chromatogr A 800 IssueID2 345–354 Occurrence Handle9561768 Occurrence Handle10.1016/S0021-9673(97)01137-0 Occurrence Handle1:CAS:528:DyaK1cXhs1Ghtrg%3D
NM Maier P Franco W Lindner (2001) ArticleTitleSeparation of enantiomers: needs, challenges, perspectives J Chromatogr A 906 3–33 Occurrence Handle11215893 Occurrence Handle10.1016/S0021-9673(00)00532-X Occurrence Handle1:CAS:528:DC%2BD3MXnvFWq
BS Moore JA Trischman D Seng D Kho PR Jensen W Fenical (1999) ArticleTitleSalinamides, anti-inflammatory depsipeptides from a marine streptomycete J Org Chem 64 IssueID4 1145–1150 Occurrence Handle10.1021/jo9814391 Occurrence Handle1:CAS:528:DyaK1MXks1Kqsg%3D%3D
K Nokihara J Gerhardt (2001) ArticleTitleDevelopment of an improved automated gas-chromatographic chiral analysis system: application to non- natural amino acids and natural protein hydrolysates Chirality 13 431–434 Occurrence Handle11466764 Occurrence Handle10.1002/chir.1057 Occurrence Handle1:CAS:528:DC%2BD3MXlvVCqsL0%3D
M Reichelt C Hummert B Luckas (1999) ArticleTitleHydrolysis of microcystins and nodularin by microwave radiation Chromatographia 49 IssueID11–12 671–677 Occurrence Handle10.1007/BF02466910 Occurrence Handle1:CAS:528:DyaK1MXksFyqu70%3D
KY Sohda M Hiramoto KI Suzumura Y Takebayashi KI Suzuki A Tanaka (2005) ArticleTitleYM-216391, a novel cytotoxic cyclic peptide from Streptomyces nobilis – II. Physico-chemical properties and structure elucidation J Antibiot 58 IssueID1 32–36 Occurrence Handle15813178 Occurrence Handle1:CAS:528:DC%2BD2MXht1Cjsrk%3D Occurrence Handle10.1038/ja.2005.3
G Thorsén J Berquist A Westlind-Danielsson B Josefsson (2001) ArticleTitleStereoselective determination of amino acids in β-amyloid peptides and senile plaques Anal Chem 73 2625–2631 Occurrence Handle11403309 Occurrence Handle10.1021/ac000861q Occurrence Handle1:CAS:528:DC%2BD3MXivFCjsro%3D
T Toyooka YM Liu (1995) ArticleTitleDetermination of D-amino acid and L-amino acid residues in peptides with fluorescent chiral tagging reagents by high-performance liquid-chromatography Chromatographia 40 IssueID11–12 645–665 Occurrence Handle10.1007/BF02315131 Occurrence Handle1:CAS:528:DyaK2MXntlars7k%3D
Author information
Authors and Affiliations
Additional information
Authors’ address: Helena Zahradníčková, Biology Centre, Academy of Sciences of the Czech Republic, Laboratory of Analytical Biochemistry, Branišovská 31, 370 05 České Budějovice, Czech Republic
Rights and permissions
About this article
Cite this article
Zahradníčková, H., Hartvich, P., Šimek, P. et al. Gas chromatographic analysis of amino acid enantiomers in Carbetocin peptide hydrolysates after fast derivatization with pentafluoropropyl chloroformate. Amino Acids 35, 445–450 (2008). https://doi.org/10.1007/s00726-007-0577-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00726-007-0577-1