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Chromatographic Separation of Amino Acids

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Applications of Ion Exchange Materials in Biomedical Industries

Abstract

Amino acids are very important in our daily life as energy sources and have several functions in metabolism since amino acids are the main key elements for the formation of proteins and peptides. All the amino acids contain a chiral carbon atom and they exist in d- and l-forms except one amino acids, i.e., glycine. Depending upon their d- or l-form, they have different biological activities in living systems. It became evident that the potential biological or pharmacological applications of amino acids are greatly restricted to the one form of the enantiomers. So, the development of methods for their separation has attracted the interest of researchers. In this chapter, we have briefly discussed the separation of amino acids with the help of different chromatographic techniques such as liquid chromatography (LC), gas chromatography (GC), thin-layer chromatography (TLC), and countercurrent chromatography (CCC). Additionally, the role of capillary electrophoresis (CE) has also been discussed toward amino acid separation. To improve the sensitivity, specificity, and applications, chromatographic techniques have been coupled with other analytical techniques, known as hyphenated techniques. In this chapter, the classification of hyphenated techniques and their roles is also discussed in addition to the advantages of chromatographic techniques over the other separation techniques. Chromatographic techniques are simple in handling, fast in the analysis, and can be used for small as well as large-scale separations with good separation efficiency and high accuracy.

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Abbreviations

β-CD:

β-cyclodextrin

(S)-NIFE:

(S)-N-(4-nitrophenoxycarbonyl)-l-phenylalanine-2-methoxyethyl ester

ABD-F:

4-aminosulphonyl-7-fluro-2,1,3-benzoxadiazol

ACC:

Adsorption column chromatography

ACN:

Acetonitrile

Ala:

Alanine

Arg:

Arginine

Asn:

Asparagine

Asp:

Aspartic acid

CCC:

Counter current chromatography

CE:

Capillary electrophoresis

CEC:

Capillary electro chromatography

CE-MS:

Capillary electrophoresis-mass spectrometry

CGE:

Capillary gel electrophoresis

CI:

Chemical ionization

CIEF:

Capillary isoelectric focusing

CITP:

Capillary isotachophoresis

CSF:

Cerebrospinal fluid

CSP:

Chiral stationary phase

Cys:

Cysteine

CZE:

Capillary zone electrophoresis

DBD-F:

4-(N,N-dimethylaminosulphonyl)-7-fluoro-2,1,3-benzoxadiazole

DEX:

Dextrin

DM-β-CD:

Heptakis (2,6-di-O-methyl)-β-cyclodextrin

EC:

Ethyl chloroformate

EI:

Electron impact ionization

ESI-MS:

Electron spray ionization-mass spectrometry

FID:

Flame ionization detection

FITC:

Fluorescein isothiocyanate

FLEC:

(+)-1-(9-lluorenyl) ethyl chloroformate

FMOC:

Fluorenylmethyloxycarbonyl chloride

GC:

Gas chromatography

GC-MS:

Gas chromatography mass spectrometry

GLC:

Gas liquid chromatography

Gln:

Glutamine

Glu:

Glutamic acid

Gly:

Glycine

GSC:

Gas-solid chromatography

His:

Histidine

HP-β-CD:

Hydroxypropyl-β-cyclodextrin

HPA-β-CD:

6-monodeoxy-6-mono (3-hydroxy)-propylamino-β-cyclodextrin

HPLC:

High performance liquid chromatography

IE-HPLC:

Ion exchange high performance liquid chromatography

Ile:

Isoleucine

IR:

Infrared spectroscopy

LC:

Liquid chromatography

LC-MS:

Liquid chromatography-mass spectrometry

LC-MS-MS:

Liquid chromatography-mass spectrometry-mass spectrometry

Leu:

Leucine

LIF:

Laser induced fluroscence

LLC:

Liquid liquid chromatography

Lys:

Lysine

Marfey’s reagent:

N-α-(2, 4-dinitro-5-fluorophenyl)-l-alaninamide

MEKC:

Micellar electrokinetic chromatography

MeOH:

Methanol

Met:

Methionine

MRM:

Multiple reaction monitoring

MS:

Mass spectrometry

MTFMPA:

Methoxy trifluoromethylphenylacetyl chloride

NAC:

N-acetyl-l cysteine

NBD:

4-fluoro-7-nitrobenzofurazan

NBD-F:

4-fluro-7-nitro-2,1,3-benzoxadiazole

NBE:

n-butyl esters

NDA:

Naphthalene dicarboxaldehyde

NMR:

Nuclear magnetic resonance

NpF:

N-heptaflurobutylryl

Npf:

N-pentafluoropropionyl

NP-HPLC:

Normal phase high performance liquid chromatography

NRMC:

N-(R)-mandelyl-(S)-cysteine

N-Tfac:

N-trifluroacetyl

OPA:

o-phthaldialdehyde

OS-γ-CD:

Octakis (2,3-dihydroxy-6-O-sulfo)-γ-cyclodextrin

PC:

Paper chromatography

PCC:

Partition column chromatography

Phe:

Phenylalanine

Pro:

Proline

RP-HPLC:

Reversed phase high performance liquid chromatography

RPPC:

Reversed phase partition chromatography

SC-HPLC:

Super critical high performance chromatography

SDS:

Sodium dodecyl sulphate

SE-HPLC:

Size exclusion high performance liquid chromatography

Ser:

Serine

SIM:

Selected ion monitoring

SLC:

Solid liquid chromatography

TEA:

Triethanolamine

TEAA:

Tetra ethyl ammonium acetate

Thr:

Threonine

TLC:

Thin layer chromatography

TLC:

Thin layer chromatography

TM-β-CD:

Heptakis (2,3,6-tri-O-methyl)-β-cyclodextrin

TOF:

Time of flight

TOFR:

Time of flight reflection

Trp:

Tryptophan

Try:

Tyrosine

UPLC-MS/MS:

Ultra-performance mass spectroscopy/mass spectroscopy

UV:

Ultraviolet

Val:

Valine

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Acknowledgements

Mr. Majhi has given the major contribution in writing this chapter along with drawing the figures and tables, taking the copyright permission, etc.

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  1. Department of Applied Chemistry, Indian Institute of Technology (Indian School of Mines), Dhanbad, 826004, Jharkhand, India

    Kartick Chandra Majhi, Paramita Karfa & Rashmi Madhuri

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  1. Kartick Chandra Majhi
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Correspondence to Rashmi Madhuri .

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  1. Chemistry Department, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia

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Majhi, K.C., Karfa, P., Madhuri, R. (2019). Chromatographic Separation of Amino Acids. In: Inamuddin (eds) Applications of Ion Exchange Materials in Biomedical Industries. Springer, Cham. https://doi.org/10.1007/978-3-030-06082-4_4

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