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Development and validation of new methods for the determination of melatonin and its oxidative metabolites by high performance liquid chromatography and capillary electrophoresis, using multivariate optimization

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Title: Development and validation of new methods for the determination of melatonin and its oxidative metabolites by high performance liquid chromatography and capillary electrophoresis, using multivariate optimization
Authors: Hevia, D.1,2, Botas, C.3, Sainz, R.M.1, Quiros, I.1, Blanco, D.3, Tan, D.X.4, Gomez-Cordoves, C.2, Mayo, J.C.1 mayojuan@uniovi.es
Source: Journal of Chromatography A. Feb2010, Vol. 1217 Issue 8, p1368-1374. 7p.
Subjects: Melatonin, Metabolites, Antioxidants, High performance liquid chromatography, Capillary electrophoresis, Multivariate analysis, Mathematical optimization, Radicals (Chemistry), Kynurenine
Abstract: Abstract: Melatonin (N-acetyl-5-metoxytriptamine, MEL) has focused a lot of attention as consequence of its multiple functions. MEL is a potent endogenous antioxidant and a free radical scavenger that reacts with several sort of radicals generating various metabolites. Two of them are N1-acetyl-N2-formyl-5-methoxykynurenine (AFMK) and N1-acetyl-5-methoxykynurenine (AMK). These compounds are important because they have also antioxidant actions as well as other important biological properties. In the present work, we develop two methods to detect and quantify these compounds (MEL, AFMK and AMK) in the same sample. For this purpose we used an experimental design, and utilized high performance liquid chromatography (HPLC-DAD) and micellar electrokinetic chromatography (MEKC) techniques with diode array detector in both of them. The limit of detection/quantification for MEL, AFMK and AMK were respectively 44/94, 18/38 and 23/51ngmL−1 by using HPLC and 13/44, 37/124 and 47/156ngmL−1 by using MEKC. This is the first time that these compounds have been separated in the same chromatogram or electroferogram. The time of analysis was faster using MEKC. Furthermore, this technique showed better resolution but HPLC offered better limit of detection and quantification for metabolites. Both methods were validated and correlation coefficients were higher than 0.999 and the range of recovery of those methods were 99.6–103.7%. Precision was evaluated as repeatability and intermediate precision with relative standard derivation <5%. When a 5μgmL−1 solution of these compounds were analyzed with both methods we do not observed any statistically significance differences. Moreover, we analyzed 3COHM (cyclic-3-hydroximelatonin), another known metabolite of melatonin, by using the same methods. The employment of these methods will offer a useful tool to contribute to answer the role of MEL, AFMK and AMK in biological system and both methods can be used in routine analysis for these compounds. [Copyright &y& Elsevier]
Copyright of Journal of Chromatography A is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: Development and validation of new methods for the determination of melatonin and its oxidative metabolites by high performance liquid chromatography and capillary electrophoresis, using multivariate optimization
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  Data: &lt;searchLink fieldCode=&quot;AR&quot; term=&quot;%22Hevia%2C+D%2E%22&quot;&gt;Hevia, D.&lt;/searchLink&gt;&lt;relatesTo&gt;1,2&lt;/relatesTo&gt;&lt;br /&gt;&lt;searchLink fieldCode=&quot;AR&quot; term=&quot;%22Botas%2C+C%2E%22&quot;&gt;Botas, C.&lt;/searchLink&gt;&lt;relatesTo&gt;3&lt;/relatesTo&gt;&lt;br /&gt;&lt;searchLink fieldCode=&quot;AR&quot; term=&quot;%22Sainz%2C+R%2EM%2E%22&quot;&gt;Sainz, R.M.&lt;/searchLink&gt;&lt;relatesTo&gt;1&lt;/relatesTo&gt;&lt;br /&gt;&lt;searchLink fieldCode=&quot;AR&quot; term=&quot;%22Quiros%2C+I%2E%22&quot;&gt;Quiros, I.&lt;/searchLink&gt;&lt;relatesTo&gt;1&lt;/relatesTo&gt;&lt;br /&gt;&lt;searchLink fieldCode=&quot;AR&quot; term=&quot;%22Blanco%2C+D%2E%22&quot;&gt;Blanco, D.&lt;/searchLink&gt;&lt;relatesTo&gt;3&lt;/relatesTo&gt;&lt;br /&gt;&lt;searchLink fieldCode=&quot;AR&quot; term=&quot;%22Tan%2C+D%2EX%2E%22&quot;&gt;Tan, D.X.&lt;/searchLink&gt;&lt;relatesTo&gt;4&lt;/relatesTo&gt;&lt;br /&gt;&lt;searchLink fieldCode=&quot;AR&quot; term=&quot;%22Gomez-Cordoves%2C+C%2E%22&quot;&gt;Gomez-Cordoves, C.&lt;/searchLink&gt;&lt;relatesTo&gt;2&lt;/relatesTo&gt;&lt;br /&gt;&lt;searchLink fieldCode=&quot;AR&quot; term=&quot;%22Mayo%2C+J%2EC%2E%22&quot;&gt;Mayo, J.C.&lt;/searchLink&gt;&lt;relatesTo&gt;1&lt;/relatesTo&gt;&lt;i&gt; mayojuan@uniovi.es&lt;/i&gt;
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  Data: &lt;searchLink fieldCode=&quot;JN&quot; term=&quot;%22Journal+of+Chromatography+A%22&quot;&gt;Journal of Chromatography A&lt;/searchLink&gt;. Feb2010, Vol. 1217 Issue 8, p1368-1374. 7p.
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  Label: Abstract
  Group: Ab
  Data: Abstract: Melatonin (N-acetyl-5-metoxytriptamine, MEL) has focused a lot of attention as consequence of its multiple functions. MEL is a potent endogenous antioxidant and a free radical scavenger that reacts with several sort of radicals generating various metabolites. Two of them are N1-acetyl-N2-formyl-5-methoxykynurenine (AFMK) and N1-acetyl-5-methoxykynurenine (AMK). These compounds are important because they have also antioxidant actions as well as other important biological properties. In the present work, we develop two methods to detect and quantify these compounds (MEL, AFMK and AMK) in the same sample. For this purpose we used an experimental design, and utilized high performance liquid chromatography (HPLC-DAD) and micellar electrokinetic chromatography (MEKC) techniques with diode array detector in both of them. The limit of detection/quantification for MEL, AFMK and AMK were respectively 44/94, 18/38 and 23/51ngmL−1 by using HPLC and 13/44, 37/124 and 47/156ngmL−1 by using MEKC. This is the first time that these compounds have been separated in the same chromatogram or electroferogram. The time of analysis was faster using MEKC. Furthermore, this technique showed better resolution but HPLC offered better limit of detection and quantification for metabolites. Both methods were validated and correlation coefficients were higher than 0.999 and the range of recovery of those methods were 99.6–103.7%. Precision was evaluated as repeatability and intermediate precision with relative standard derivation &lt;5%. When a 5μgmL−1 solution of these compounds were analyzed with both methods we do not observed any statistically significance differences. Moreover, we analyzed 3COHM (cyclic-3-hydroximelatonin), another known metabolite of melatonin, by using the same methods. The employment of these methods will offer a useful tool to contribute to answer the role of MEL, AFMK and AMK in biological system and both methods can be used in routine analysis for these compounds. [Copyright &amp;y&amp; Elsevier]
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  Data: &lt;i&gt;Copyright of Journal of Chromatography A is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder&#39;s express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.&lt;/i&gt; (Copyright applies to all Abstracts.)
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RecordInfo BibRecord:
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      – Type: doi
        Value: 10.1016/j.chroma.2009.12.070
    Languages:
      – Code: eng
        Text: English
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      Pagination:
        PageCount: 7
        StartPage: 1368
    Subjects:
      – SubjectFull: Melatonin
        Type: general
      – SubjectFull: Metabolites
        Type: general
      – SubjectFull: Antioxidants
        Type: general
      – SubjectFull: High performance liquid chromatography
        Type: general
      – SubjectFull: Capillary electrophoresis
        Type: general
      – SubjectFull: Multivariate analysis
        Type: general
      – SubjectFull: Mathematical optimization
        Type: general
      – SubjectFull: Radicals (Chemistry)
        Type: general
      – SubjectFull: Kynurenine
        Type: general
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      – TitleFull: Development and validation of new methods for the determination of melatonin and its oxidative metabolites by high performance liquid chromatography and capillary electrophoresis, using multivariate optimization
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              M: 02
              Text: Feb2010
              Type: published
              Y: 2010
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