Quantitative determination of amino acids in earthworm meal (Eisenia andrei) by a Surveyor HPLC system in conjunction with pre-column 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate derivatization.

Authors

  • José Fernando Ovalles Departamento de Ciencias de los Alimentos, Facultad de Farmacia y Bioanálisis, Universidad de Los Andes, Mérida (Venezuela).
  • Ana Luisa Medina Departamento de Ciencias de los Alimentos, Facultad de Farmacia y Bioanálisis, Universidad de Los Andes, Mérida (Venezuela).
  • Elil Márquez Departamento de Ciencias de los Alimentos, Facultad de Farmacia y Bioanálisis, Universidad de Los Andes, Mérida (Venezuela).
  • Julie Rochette Programa de Cooperación Postgraduados, Franco-Venezolano, Universidad de Montpellier, Montpelier (Francia).
  • José Rafael Luna Departamento de Toxicología, Facultad de Farmacia y Bioanálisis, Universidad de Los Andes, Mérida (Venezuela).
  • Marielba Morillo Departamento de Ciencias de los Alimentos, Facultad de Farmacia y Bioanálisis, Universidad de Los Andes, Mérida (Venezuela).

Keywords:

6-aminoquinolyl-N-hydroxysuccinimidyl carbamate, AQC, earthworm meal, HPLC

Abstract

Aim: The objective of this study was to evaluate the integration of the AccQ•Tag derivatization system with the Finnigan Surveyor Plus HPLC system to determine the amino acids (aa) composition of earthworm meal protein post-hydrolysis.

Materials and Methods: In lab cultivated earthworms (Eisenia andrei) were reduced to flour which was then hydrolyzed with 6M HCl at 110 °C for 24 hours in a closed system. The hydrolysis product was neutralized and their aa were derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). The derivatized-aa were separated by RP-HPLC and detected by fluorescence.

Results and Conclusion: The proposed integration makes optimal use of both the modular design of the Surveyor Plus HPLC for versatility and flexibility and the main features of the AccQ•Tag derivatization system in terms of stability and reproducibility. Analytical validation parameters were studied both before and after derivatization with AQC. The resulting data were within acceptable ranges for this type of analysis. Pre-column derivatization with AQC yielded appropriate sensitivities within the low pmol range per injection. Earthworm meal generated the following aa; the most abundant (w/w) being: Glu, Asp, Arg, Leu, and Lys (4 % - 10 %), whereas the lowest content corresponded to Met (< 1. 5%), which is comparable to fishmeal. The analytical proposal can be used with confidence in earthworm meal quality control to guarantee the appropriate aa content to create an optimum fish diet.

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Published

2014-09-20

How to Cite

1.
Ovalles JF, Medina AL, Márquez E, Rochette J, Luna JR, Morillo M. Quantitative determination of amino acids in earthworm meal (Eisenia andrei) by a Surveyor HPLC system in conjunction with pre-column 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate derivatization. Ars Pharm [Internet]. 2014 Sep. 20 [cited 2024 Jul. 22];55(3):35-44. Available from: https://revistaseug.ugr.es/index.php/ars/article/view/4520

Issue

Vol. 55 No. 3 (2014)

Section

Original Articles

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