So your question has two parts, the first asks about d-ROM's and the other about the FRAS 4 EVOLVO P. First, the d-ROM's test at its basic form appears to be a decent assay for measuring any type of hydroperoxide (hydrogen peroxide, lipid peroxides, etc). It is based on Fenton chemistry, where a hydroperoxide will react with iron to form an actual radical specie (remember, most hydroperoxides are not radicals, but just reactive species). This newly formed radical specie reacts with a colorometric substrate, and you get a readout on a spectrophotometer. While I have read about this assay, to be completely honest I have not seen it much in the literature.

As for the FRAS 4 EVOLVO P, well, I am not sure that is a device you want to invest in. After reading about the machine a bit, it is clear that it can only do very rudimentary and not well accepted measures of "reactive oxygen species."  Many of the assays it runs do not even measure radicals.

I would stick to the established protocols such as electron paramagenetic resonance, fluorogenic probes or proteins, or even assays like TBARS.  While each of these is not perfect, they are much more established and studies than what you have listed above. This will help you immensely down the line when you want to publish. 

Good luck,

~Adam

In Indeed FRAS 4 EVOLVO P is not the better option to measure oxidative stress, but we have a huge problem, our samples have been stored for 7 years at -80º were taken without caution and were thawing once at least. To think in other methodologies is difficult for this conditions, and for this reason I would like to know if FRAS 4 EVOLVO P we will allow obtain any result about oxidative stress.

http://www.fras4evolvo.com/fras4evolvo.php

Thank you Adam,

Hi Mayely, I want to ask what kind of samples do you have? I have been  using the Diacron’s products for three years, generally on fresh samples and sometimes on frozen ones, too. The test is fairly recent and literature is not large but, as Adam correctly says, if you only perform this you will have partial information on the oxidative status of your sample. You should also take into account the storage period,  the effect of thawing and the short self-life of radical species since all these factors could influence  the samples quality.

Also, I have experienced in working  with both one of the Diacron automatized system  and a normal spectrophotometer. Thus, in this case I don’t suggest you to buy the system because, being a colorimetric reaction (read at 505nm) you can only buy the kit and use the photometer of your lab for the measurement. The kits are available in different formats  depends on the number of your samples, they also include a calibrator and you can perform both an endpoint or kinetic measurement.

I hope that the above information will be useful but if you have any further questions please email me.

Sincerely

Barbara

Barbara,

I have serum samples (no plasma) in these contitions. Your recommendation seems to be a best option, I have an spectrophotometer with these characteristics (Analyzer A15, Biosystems), I will try it.

Thank you very much!

Dear Colleague,

as I have just indicated in an answer to anothder of your question, the main problem resides in the samples themselves: no samples stored in the way you described can support you with reliable results. In this sense there is no existing assay that can improved the compromised features of your samples.

In addition, d-ROM is probably the less reliable among all the available methods to assess oxidative stress. I do not recommend it at all.

 I tested this method more than ten years ago, and concluded that as stated by Maurizio Battino, that  "is probably the less reliable among all the available methods to assess oxidative stress". Pleased note that measuring oxidative stress is not an easy task for anyone and that does not exists and easy method available for all samples and biological matrix. We have summarized the related problems in the review:

Oxidative stress and human diseases: Origin, link, measurement, mechanisms, and biomarkers. http://www.ncbi.nlm.nih.gov/pubmed/19958214

However dROMs test meets the needs of many researchers to obtain an easy method to say "we detect oxidative stress", in that it generates numbers by an easy procedure. Note that the values are expressed as arbitrary units, i.e. Carr units. If you calculate the concentration expressed as H2O2 content they correspond to 20-30 mM in serum or plasma...

The claim that you can just eat glutathione, can be likened to saying that all you have to do to get smart is eat brain tissue supplements. Glutathione is degraded into its three constituent AA, in the gut i.e. Cys, Glu and Gly, which are by the way conteined in all protein rich tissues. GSH treatment is not proven to be of any benefit, it is just an expensive method to try to increase cellular glutathione. It is more logical to use the less expensive NAC (N-acethyl cysteine), but also in this case, it is not proven that this can increase cell GSH. On my experience NAC can barely have an effect at very high concentrations.

for a long time  my opinon  ,the d-ROMs test measures serum ceruloplasmine levels!...........

 maybe usuful following article...

Free Radic Res. 2014 Aug;48(8):883-9. doi: 10.3109/10715762.2014.919390. Epub 2014 May 21.

Assessment of oxidative stress in serum by d-ROMs test.

Kilk K1, Meitern R, Härmson O, Soomets U, Hõrak P.

Abstract

Assessment of oxidative stress is an important but technically challenging procedure in medical and biological research. The reactive oxygen metabolites (d-ROMs) test is a simple assay marketed for analyzing the total amount of hydroperoxides in serum via the Fenton's reaction. Earlier reports have raised a suspicion that a part of the signal detected in the assay comes from sources other than metabolites generated by oxidative stress. The aim of this study was to identify which serum components interfere with the d-ROMs signal. By application of sodium azide, ethylenediaminetetraacetic acid, sodium dodecylsulphate, varying temperature, and spiking endogenous substances we demonstrate that in the case of mammalian sera the assay determines ceruloplasmin (CP) activity with potential interferences from hydroperoxides, iron level, thiols, and albumin. In sera of avian species hydroperoxides contribute more to the test outcome, but the CP part is insensitive to inhibition by azide. In conclusion, this assay has deficiencies in terms of detecting realistic concentrations of hydroperoxides, is mostly measuring CP and is also interfered with other serum components, making it very difficult to interpret in most biological systems.

Dear Mayely,

I noticed your discusssion only recently. We are measuring dROMs now for 5 years in about 20.000 samples on our clinical analyzer with great succes. We found a correlation with several chronic diseases (cancer CVD) and aging. This was recently published in a number of papers and more have been submitted. In serum the assay is very stable. We did a study on short time (already published) and for long time in which we keep the serum samples frozen for several years and the assay is perfectly stable. One paper about the 1-year stability will be published soon since I have the page proofs now for correction (Biomarkers in Medicine). Last year October 2014 there was a conference at your university on Biogerontology and there I presented also some results on dROMs.

If you want to do the assay in a spectrophotometer you must pay much attention to the QC of the assay and use each day a batch of frozen serum as control serum.

Regards

Eugène

In line with the comments provided by Eugene, I have used these kits for 10 years and they produce reliable and consistent results.

Dear Mayely,

I also had the same experience as Eugene and Pietro.

The paper "Assessment of oxidative stress in serum by d-ROMs test." is plenty of mistakes, it suffers of serious methodological inadequacy. I just wonder why the reviewers did not find them, they are so evident!

cheers

David

Hi Mayely

have a look at this recent study, which once again shows that the d-ROMs assay

is a very good test for the assessment of oxidative stress.

http://www.tandfonline.com/doi/abs/10.3109/10715762.2015.1136063

Cheers

David

Yes, of course!!!

I have no doubts that who patented d-ROMs assay now write it is good. Mr Carratelli, is one of the author and do you remember how have been called the measuring units of dRom-assay???

Just in case, I give you the answer: "Carratelli Units"......:-)

It seems to me that usually this is called C.O.I.: Conflict of Interest

For your information this assay has also been successfully validated by other laboratories. The few critics raised so far were based on methodological inadequacy.

Just like questioning the validity of the Gauss theorem on magnetic flux only because magnetic fields happen to be measured in Gauss (now Tesla).

One should deal with facts rather than with names, and facts are that hundreds of studies have been carried out successfully using the d-ROMs test.

The problem with this test is its wide distribution akso to a large cohort of non-experts or even non-MD. So people marketing-wise oversimplifying a far more complex redox balance. Wannabe antiaging doctors among the worst preachers.