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Determination of Cortisol,testosterone ratios in saliva using

HPLCBy Sirish Narramaneni23rdJuly 2009

Supervisor

Birthe Nielsen

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Objectives:

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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Biomarkers

Indicator:

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

glands.

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

adrenals.

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

mind.

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 http://www.macses.ucsf.edu/Research/Allostatic/notebook/FAQs-salivcort.pdfTechnical Issues by Clemens Kirschbaum, Ph.D.

y

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

Nishigawara, Okayama 703-8516, Japan.