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Development and Validation of a HPTLC Method for Simultaneous Estimation of L-Glutamic Acid and γ-Aminobutyric Acid in Mice Brain.

Sancheti JS, Shaikh MF, Khatwani PF, Kulkarni SR, Sathaye S - Indian J Pharm Sci (2013)

Bottom Line: A new robust, simple and economic high performance thin layer chromatographic method was developed for simultaneous estimation of L-glutamic acid and γ-amino butyric acid in brain homogenate.The high performance thin layer chromatographic separation of these amino acid was achieved using n-butanol:glacial acetic acid:water (22:3:5 v/v/v) as mobile phase and ninhydrin as a derivatising agent.The limit of detection and limit of quantification for L-glutamic acid was found to be 10 and 20 ng and for γ-amino butyric acid it was 4 and 10 ng, respectively.

View Article: PubMed Central - PubMed

Affiliation: Departments of Pharmaceutical Science and Technology, Insitute of Chemical Technology, N. P. Marg, Matunga, Mumbai-400 019, India.

ABSTRACT
A new robust, simple and economic high performance thin layer chromatographic method was developed for simultaneous estimation of L-glutamic acid and γ-amino butyric acid in brain homogenate. The high performance thin layer chromatographic separation of these amino acid was achieved using n-butanol:glacial acetic acid:water (22:3:5 v/v/v) as mobile phase and ninhydrin as a derivatising agent. Quantitation of the method was achieved by densitometric method at 550 nm over the concentration range of 10-100 ng/spot. This method showed good separation of amino acids in the brain homogenate with Rf value of L-glutamic acid and γ-amino butyric acid as 21.67±0.58 and 33.67±0.58, respectively. The limit of detection and limit of quantification for L-glutamic acid was found to be 10 and 20 ng and for γ-amino butyric acid it was 4 and 10 ng, respectively. The method was also validated in terms of accuracy, precision and repeatability. The developed method was found to be precise and accurate with good reproducibility and shows promising applicability for studying pathological status of disease and therapeutic significance of drug treatment.

No MeSH data available.


Related in: MedlinePlus

HPTLC desitogram of standards.HPTLC chromatogram of standard L-glutamic acid and GABA at 40 ng/spot concentration.
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Figure 2: HPTLC desitogram of standards.HPTLC chromatogram of standard L-glutamic acid and GABA at 40 ng/spot concentration.

Mentions: Linearity of any analytical method defines its ability within a given range. The test results thus obtained are directly or through mathematical transformation, proportional to the concentration of analyte[13]. This developed method was found to be linear for L-glutamic acid and GABA in a concentration range of 10-100 ng/spot for each amino acid (figs. 1 and 2). Fig. 1, displays three dimensional overlay of HPTLC densitograms of the calibration spots of L-glutamic acid and GABA at 550 nm. The regression data are reported in Table 1.


Development and Validation of a HPTLC Method for Simultaneous Estimation of L-Glutamic Acid and γ-Aminobutyric Acid in Mice Brain.

Sancheti JS, Shaikh MF, Khatwani PF, Kulkarni SR, Sathaye S - Indian J Pharm Sci (2013)

HPTLC desitogram of standards.HPTLC chromatogram of standard L-glutamic acid and GABA at 40 ng/spot concentration.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3928736&req=5

Figure 2: HPTLC desitogram of standards.HPTLC chromatogram of standard L-glutamic acid and GABA at 40 ng/spot concentration.
Mentions: Linearity of any analytical method defines its ability within a given range. The test results thus obtained are directly or through mathematical transformation, proportional to the concentration of analyte[13]. This developed method was found to be linear for L-glutamic acid and GABA in a concentration range of 10-100 ng/spot for each amino acid (figs. 1 and 2). Fig. 1, displays three dimensional overlay of HPTLC densitograms of the calibration spots of L-glutamic acid and GABA at 550 nm. The regression data are reported in Table 1.

Bottom Line: A new robust, simple and economic high performance thin layer chromatographic method was developed for simultaneous estimation of L-glutamic acid and γ-amino butyric acid in brain homogenate.The high performance thin layer chromatographic separation of these amino acid was achieved using n-butanol:glacial acetic acid:water (22:3:5 v/v/v) as mobile phase and ninhydrin as a derivatising agent.The limit of detection and limit of quantification for L-glutamic acid was found to be 10 and 20 ng and for γ-amino butyric acid it was 4 and 10 ng, respectively.

View Article: PubMed Central - PubMed

Affiliation: Departments of Pharmaceutical Science and Technology, Insitute of Chemical Technology, N. P. Marg, Matunga, Mumbai-400 019, India.

ABSTRACT
A new robust, simple and economic high performance thin layer chromatographic method was developed for simultaneous estimation of L-glutamic acid and γ-amino butyric acid in brain homogenate. The high performance thin layer chromatographic separation of these amino acid was achieved using n-butanol:glacial acetic acid:water (22:3:5 v/v/v) as mobile phase and ninhydrin as a derivatising agent. Quantitation of the method was achieved by densitometric method at 550 nm over the concentration range of 10-100 ng/spot. This method showed good separation of amino acids in the brain homogenate with Rf value of L-glutamic acid and γ-amino butyric acid as 21.67±0.58 and 33.67±0.58, respectively. The limit of detection and limit of quantification for L-glutamic acid was found to be 10 and 20 ng and for γ-amino butyric acid it was 4 and 10 ng, respectively. The method was also validated in terms of accuracy, precision and repeatability. The developed method was found to be precise and accurate with good reproducibility and shows promising applicability for studying pathological status of disease and therapeutic significance of drug treatment.

No MeSH data available.


Related in: MedlinePlus