I suggest to use HPLC with electrochemical detection. with that you can measure al monoamines, DA, NE, Epi, DOPAC and 5-HT.

Im looking for same thing :) however with no luck...methods are realy not easy and far from economic! Realy I would gladly whant to hear someones ideas...If you whant i could send you some methods where fluorescent detection is used...

Dear sir,

I found an abstract for determination of dopamine,

but I have doubt about its accuracy. Please give me suggestion about authenticity of this method, whether it is reliable or not.

Find abstract for same..and give me your valuable suggestion.

Author information:

Guo L1, Zhang Y, Li Q.

Abstract

In the present work, we developed a simple, sensitive and inexpensive method to determine dopamine hydrochloride using potassium ferricyanide-Fe(III) by spectrophotometry. The results show that Fe(III) is deoxidized to Fe(II) by dopamine hydrochloride at pH 4.0, and then Fe(II) reacts with potassium ferricyanide to form a soluble prussian blue (KFe(III)[Fe(II)(CN)6]). The absorbance of this product was monitored over time using a spectrophotometer at an absorption maximum of 735 nm, and the amount of dopamine hydrochloride could be calculated based on the absorbance. A good linear relationship of the concentration of dopamine hydrochloride versus absorbance was observed, and a linear regression equation of A = 0.022 + 0.16921C (microg mL(-1)) was obtained. Moreover, the apparent molar absorption coefficient for the indirect determination of dopamine hydrochloride was 3.2 x 10(4) L mol(-1) cm(-1). This described method has been used to determine dopamine hydrochloride in pharmaceutical, banana, urine and serum samples with satisfactory results.

Well I can;t say if the method is good/bad because its not my area...I see that chemicals are cheap so you can try it easy...from my point of view i would/will definitively try it!

If you want reliable results then use Microbore HPLC with amperometric detector. Otherwise it will be a waste of time.

Hello Dr. Tirgar, exactly wich monoamines are you trying to identify? What will be the source of your sample? are you interested in determining only one or multiple compounds?. Best regards

Thank you very much too all  for your valuable suggestions.

Actually I am looking forward to detect effect of a research drug on mono-amine neurotransmitters like Adrenaline, Noradrenaline, Dopamine, 5HT and also for insulin, thyroid hormones on experimental animal blood sample or brain homogenate.

Kindly guide me....

Well there are a lot of methods that suggest HPLC with different detectors that are not available (and not cheep)  to everyone thus I also want to  hear about methods that use spectroscopy (UV-VIS) or fluorescent detection.... 

Perhaps, to collaborate with a lab that has the instrumentation and experience? I recommend prof Sarre from the VUB.

I suggest to use HPLC with electrochemical detection. with that you can measure al monoamines, DA, NE, Epi, DOPAC and 5-HT.

One of the earliest HPLC assays for simultaneous determination of monoamines by HPLC published from the dept. of physiology, Univ. oF Saskatchewan College of Med, Saskatoon, SK, Canada. But I cannot locate that publication. Here is a citation based on that assay:

Liu YL, Cheng AT, Chen HR, Hsu YP. Simultaneous HPLC of twelve monoamines and

metabolites shows neuroblastoma cell line releases HVA and HIAA. Biomed

Chromatogr. 2000 Dec;14(8):544-8. PubMed PMID: 11113939.

HPLC with electrochemical detection allows the dectection of monoamines in biological samples. Extraction is very easy and the results highly reproducible.

I agree with Ali Abdil Razzaq Muhammed Noori Aldallal. Indeed, an adequate method is chromatography. I mean, HPLC. But the precise definition of neurotransmitters by the method requires highly skilled researchers. There are a variety of necessary control procedures that are required for accurate determination of neurotransmitters. All the best.

You can estimateall the catecholamines in the same sample by RIA method (vide may publications)

It depends on the circumstances. Objectives of work, staff development, data mapping literature, a comparison of the two methods, and much more. Selecting or RIA or HPLC - tandem mass spectrometry is discussed in the articles (see references).

Yamane R., Nakamura T., Matsuura E., Ishige H., Fujimoto M. A simple and sensitive radioimmunoassay for adenosine. J. Immunoassay. 1991. Vol. 12, No 4. P. 501-519.

Francis R.C., Pickerodt P.A., Salewski L., Boemke W., Höhne C. Endocannabinoids in chronic migraine: CSF findings suggest a system failure. Vet. J. 2010. Vol. 183, No 2. P228-231. doi: 10.1016/j.tvjl.2008.10.008.

Thank you very much for your valuable suggestions which really help in my research for proper and accurate determination of catecholamine and other neurotransmitter in biological sample..

Hope if you have any further suggestion please guide me 

thank you to all

HPLC method by the skilled technician/researchers is very effective in determining catecholamines.

HPLC with electrochemical detection could be useful. I agree wit both  Silvia and Teresa adn of course with  Sultan.

Good luck.

I believe that all the recommendations are very useful for performing precision studies.

Why don't you use the very pwerful method consisting in electrochemical detection (HPLC) that will enable you to measure very precisely a lots of  neurotransmitter compounds ?

You could use enzyme based microelctrode array...

Artile name:

Resting glutamate levels and rapid glutamate transients in the prefrontal cortex of the Flinders Sensitive Line rat: a genetic rodent model of depression.

Link:

http://www.ncbi.nlm.nih.gov/pubmed/21525860

If you don't get consistent or low enough levels of detection with HPLC try LC-MS or perhaps, more easily, an ELISA-based method.

Hello Teresa

would you please provide me sample protocol for neurotransmitter estimation from Brain homogenate by HPLC-ECD method.. I am struggling to extract the neurotransmitter from from homogenate..

      Thanks in advance for your guidance. 

Hi,

Please click on the attached link below! I hope this information will help you!

Best Regards

http://link.springer.com/article/10.1007%2FBF02490443

Hello,

HPLC-ECD methods is an easy and accurate method to detect monoamines.

Best high throughput, sensitive method is HPLC with electrochemical detection. But to the points raised above, this is not easy to deploy if you have no experience and the equipment is also very expensive. I am the Director of an HPLC Core Lab and have a long history of collaborating with investigators to help them analyze monoamines from a variety of sources - plasma, brain tissue, microdialysate. My expertise is not limited to monoamines, the range of compounds that can be quantified with HPLC methods is vast. Please contact me if you are interested - [email protected]

Impressive question that provoked very valuable comments and informations from our RG colleagues

Following

HPLC Method can be used to estimate the neurotrnsmitter...............

Dopamine Levels In The Brain Striatum by LC-MS/MS.

ØThe straitum was homogenized with the addition of Phosphate buffers. Homogenate was acidified with 50 μL of formic acid (98-100%, J.T Baker, USA), the Dopamine were extracted from the supernatant by solid-phase extraction using mixed-mode strong cation-exchange and reversed-phase cartridges. ØThe dopamine were eluted with 10 mL of methanol containing 2% ammonia. To the collection tubes, 2 mL of methanol and 750 μL of formic acid were added to neutralize the ammonia used in the elution. ØDopamine was fractionated from the residue with an Agilent HP 1200 liquid chromatography (Hewlett-Packard GmbH, Waldbronn, Germany) equipped with a quaternary pump, an autosampler, a column compartment, detected by Electro chemical detector.