Fluorimetric analysis catecholamines is widely used.The following references may help you to set up your analysis ( simple HPLC and HPTLC methods are also known)

based on availability of instruments at your lab.

1.Measurement of catecholamines and their metabolites

193.146.160.29/gtb/sod/usu/

by RT Peaston - 2004 - methods for the biochemical detection of catecholamine-secreting tumours. Ann Clin Biochem ..... procedures that allow the rapid and simple measure- ment of ...

2.A simple, rapid HPLC method for the concurrent measurement of ...

www.ncbi.nlm.nih.gov/pubmed/RJ Carey - 1995 -

A simple, rapid HPLC method for the concurrent measurement of cocaine and catecholamines in brain tissue samples. Carey RJ, DePalma G. VA Medical ...

3.Simple, rapid and sensitive determination of epinephrine and ...

www.ncbi.nlm.nih.gov/pubmed/11833678by J Westermann - 2002 -

METHODS: A non-competitive enzyme-linked immunosorbent assay (CatCombi ELISA) kit for the measurement of epinephrine and norepinephrine concentrations in ... plasma is required for duplicate determinations of both catecholamines.

]

4.Analytical methods for determining urinary catecholamines in ...

144.206.159.178/ft/38/183876/4698610.

by RPH Nikolajsena - 2001 -

usefulness of the method for measurement of catecholamines in urine from healthy subjects. ..... hence, this detection method was a very simple one; and Said et ...

Having measured CATs using HPLC and ELISA I would recommend ELISA every time. You do need quite a lot of plasma, however, for ELISA 200-300ul. Therefore, this can be a problem when working with small animals.

ELISA kits can also be used but the cost factor (500-600 USD per kit) and on time availability makes the researcher to look for economical and equally reproducible methods (1-2 USD per test) .

we have different techniques for stress measurement including cathecolamine. May I ask you to contact directly our laboratory at [email protected] ( www.saistmp.com)?.

Thank you all as every answer has helped me

Dear Dr. Samina, I may suggest also to read the interesting article in Theoretical Biology and Medical Modeling , 2005,2:11,1-10, entitled A topological model of biofeedback based catecholamine interactions. by T.K. Basak,S.Halder,M. Kumar.R.Sharma.B.Midya. As I see in our laboratory it gives important and rapid informations.

If I can add to the discussion on cortisol, I am just submitting papers on the use of salivary cortisol as a stress marker in children with Williams syndrome and in actors. Spot salivary cortisol is of absolutely no value. However, looking at individuals pattern of salivary cortisol level (ie it should be highest in the morning upon waking, lowest at bedtime before sleep) you can get an indication of stress. During the day cortisol goes up and down in acute responses to circumstance and is not an indication of chronic stress leading to clinical mental health - Wellbeing symptoms.

Thus, cortisol has to be measured as a pattern and interpreted in light of psychophysiology I.e high bedtime cortisol is associated with the laying down of emotional memories which can be positive or negative depending on the stress experienced.

However, an absolute measure of urinary adrenaline metabolites might give a spot indication of stress load. Indeed very very high levels might correlate with clinical manifestations of chronic stress.

I look forward to a heated debate now.

Dr Ray Iles:

Trust you have considered the metabolites of Cortisol also for your study on quantification of total Cortisol.

Needless to mention you have identified and noted the exact stages of stress your subjects are in at the time of sample collection ( repulsion , adaptation and succumbing stages of stress conditions....Dr.Hans Selye models) which will provide more scientific data for understanding treatment schedules. We look forward for your publications.

Dear Dr B K Chakravarthy I had not considered metabolites of Cotisol as being in Saliva - thinking salivary levels being a simple transdudate of blood can you point me to some papers? I am planning a simple Cortisol ELISA because of costs and resources available. ( and I will look at Dr Hans Selyle models Very helpful to me and I think the original questioner)

Dr.Ray Iles:

Please find some useful references :

No idea which stress model you are using but with my past experiences with "Under nourishment during weaning period in mammalian species.... has effect on cortisol levels...and has a great importance on the existing senerio in most of the countries. If you have some data we look forward for your research work on that.

1.Metabolism of adrenal steroids

www.uptodate.com/contents/metabolism-of-adrenal-steroids28 Sep 2012 – Reduction reactions can also result in “regeneration” of cortisol from its inactive ... has been identified [8] and detailed studies of a 13-year-old patient with a ... total absence of 5 beta reduced metabolites of cortisol and cortisone in urine [9]. ... Always seek the advice of your own physician or other qualified ..

2.Quantification of cortisol, cortisol immunoreactive metabolites, and ...

www.ncbi.nlm.nih.gov › ... › Can J Vet Res › v.70(2); Apr 2006Shareby C Rehbinder - 2006 -

3.Hormones as indicators of stress

www.depts.ttu.edu/animalwelfare/classes/...by E Möstl - 2002

Dear Dr. Ray Kruse Iles ,

You mention that "an absolute measure of urinary adrenaline metabolites might give a spot indication of stress load. Indeed very very high levels might correlate with clinical manifestations of chronic stress"

Could you, please, share your ideas if urinary, plasma or, maybe better, salivary metanephrine may reflect acute or chronic stress?

Thank you