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  • 8/6/2019 Sirish Presentation


    Determination of Cortisol,testosterone ratios in saliva using

    HPLCBy Sirish Narramaneni23rdJuly 2009


    Birthe Nielsen

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    y To determine the ratio of Cortisol and Testosterone in

    individuals so that we can detect a state of mind of aindividuals.

    y In a stress induced individuals, cortisol levels are high.

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    y Identify the progress of the disease.y Indicates presence of certain organism including its

    history of presence even if they no longer exist.

    Example: Antibody .

    Blood, Saliva, urine and sweat contain biomarkers.

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    Applicationsy Immunophenotyping

    y Bacteria analysis

    y Cell cycle analysisy DNA analysis

    y Gene expression

    y Lineage analysis

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    Salivay It is a water like substance secreted from salivary


    Advantages of saliva selection:

    y Easy to collect

    y Non invasive

    y No trained professional required for collection

    Collection Using Salivette tubes

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    Cortisoly Most potent glucocorticoid produced by human


    y Synthesized from chloesterol and its productionstimulated by pituitary ACTH regulated CRF.

    yAlso known as Stress hormone.

    y Normal range:

    - For men 10 ng/ml- For women 8 ng/ml

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    Testosteroney Primary male sex hormone.

    y Synthesized by leydigs cells in testis at varying

    amounts.y Functions: Maintenance of secondary sexual

    characteristic of males.

    y Hypogonadism: Detected using salivary testosterone

    (Mornings).y Normal ranges: 500-1000 ng/dL.

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    Cortisol ,Testosterone Ratioy These levels alter depending on individual state of


    y Usually during nights cortisol levels are generally lowcompared to testosterone levels.

    y Example: Athletes.

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    HPLCy High performance liquid chromatography is also

    called high pressure liquid chromatography since high

    pressure is used compared to other chromatography.y The principle involved in HPLC is adsorption.

    y High performance Liquid Chromatography is used forthe analysis of mixture of compounds. It is used for

    identification, separation and purification of drugs.

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    Schemeofworky Phase I

    Extensive literature search

    y Phase II

    Procurement of materials.

    Preparation of stock solutions.

    y Phase III

    Analysis using HPLC

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    Referencesy SalivaryTestosterone: A Reliable Approach to the

    Diagnosis ofMale HypogonadismAlejandro L. Arregger; Liliana N. Contreras; Omar R.Tumilasci; Daniel R. Aquilano; EstelaM. L. Cardoso.

    y Issues by Clemens Kirschbaum, Ph.D.


    Determination of cortisol in human saliva by HiroyukiKataoka , ErikoMatsuura and KurieMitaniSchool of Pharmacy, Shujitsu University,

    Nishigawara, Okayama 703-8516, Japan.