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Enantioselective micro‐2D‐HPLC determination of aspartic acid in the pineal glands of rodents with various melatonin contents
Author(s) -
Han Hai,
Miyoshi Yurika,
Oyama Tsubasa,
Konishi Ryoko,
Mita Masashi,
Hamase Kenji
Publication year - 2011
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201100609
Subject(s) - pineal gland , melatonin , derivatization , high performance liquid chromatography , enantioselective synthesis , chromatography , chemistry , aspartic acid , medicine , biology , biochemistry , amino acid , endocrinology , catalysis
Enantioselective determination of aspartic acid (Asp) in the pineal gland of rodents with various melatonin contents was performed using a highly sensitive and selective two‐dimensional HPLC system. After derivatization of the amino group with 4‐fluoro‐7‐nitro‐2,1,3‐benzoxadiazole (NBD‐F), NBD‐Asp was separated using a capillary monolithic ODS column in the first dimension. The fraction of NBD‐Asp was automatically collected and transferred to the second dimension, and the D ‐ and L ‐Asp were separated and determined using a narrowbore enantioselective column. Large amounts of D ‐Asp were observed in the pineal gland of the rats and specific strains of mice (C3H and CBA) possessing a high concentration of melatonin in their pineal gland. On the other hand, the amounts of D ‐Asp were small in the pineal gland of mice possessing a trace or no melatonin in their pineal gland (ddY, ICR, C57BL and BALB/c). In other tissues and physiological fluids, no significant strain‐dependent changes of the D ‐Asp amounts were observed. These results indicate that large amounts of D ‐Asp are present only in the pineal gland containing large amounts of melatonin, and special care should be taken when selecting mouse strains for the investigation of D ‐Asp.