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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2017: 100.00
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2017: 100.00
For more details click here

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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2017: 100.00
NAAS RATING 2018: 5.38

Int.J.Curr.Microbiol.App.Sci.2018.7(2): 1397-1404
DOI: https://doi.org/10.20546/ijcmas.2018.702.168


Simultaneous Determination of Fish Steroidal Hormones using RP-HPLC with UV Detection by Multi-step Gradient Elution Technique
Ningthoukhongjam Soranganba* and I.J. Singh
Department of Fisheries Resource Management, College of Fisheries G.B.P.U.A & T, Pantnagar, Uttarakhand, India - 263145
*Corresponding author
Abstract:

A reversed phase high performance liquid chromatography (HPLC) method with UV detection by multi-step gradient elution method has been developed for simultaneous determination of testosterone, 17β-estradiol, 17α, 20β-dihydroxy-4-pregnen-3-one and cortisol in fish. The method described includes limited sample preparation with single step SPE using C¬¬¬¬¬18 cartridge. A multi-step gradient elution of water (A) and acetonitrile (B) with flow rate 1-1.5ml/min with curve index 5of 20% to 80% of B at various intervals and variable detection at UV max of 203nm and 242nm. Validation of the method was performed in terms of recovery with 87.2-106%, intra- and inter-assay variability of 0.93-21.28% and 0.40-9.18% respectively and linearity between 10-1200ng/ml. E2 shows the least LOD (25.16ng/ml) and LOQ (229.8ng/ml) followed by cortisol (45.64ng/ml and 138.32ng/ml) T (72.946ng/ml and 221.04ng/ml) and DHP (75.83ng/ml and 138.32ng/ml). The developed chromatograph was applied to analyse plasma hormonal level in amur carp.


Keywords: RP-HPLC, Testosterone, 17β-estradiol, 17α, 20β-dihydroxy-4-pregnen-3-one (DHP), Cortisol, amur carp, Multi-step gradient
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How to cite this article:

Ningthoukhongjam Soranganba and Singh, I.J. 2018. Simultaneous Determination of Fish Steroidal Hormones using RP-HPLC with UV Detection by Multi-step Gradient Elution Technique.Int.J.Curr.Microbiol.App.Sci. 7(2): 1397-1404. doi: https://doi.org/10.20546/ijcmas.2018.702.168