Pyrrole Test

Urinary Pyrrole Test? Some quick facts.

The urinary pyrrole fraction occurs in the urine when the body is under stress. The test results indicate to your practitioner the stress severity. The associated mental health issues are related to the stress. Repairing the stress should lower the pyrrole and reduce mental health issues.

Symptoms are related to the individual’s biochemistry and the underlying cause of the stress.

Elevated pyrroles (aka “pyroluria”) have not been proven to be genetic or a disorder.

The urinary pyrroles measure has no purpose unless it is scientifically proven to be useful in improving outcomes for patient care.

A high level of urinary pyrroles, when combined with clinical signs of mental health issues, can give the patient hope that the problem can be resolved and a return to normal health is possible.

Urinary pyrroles are waste products so it must be collected and transported correctly to prevent the product disintegrating. Read further about Pyrrole Testing Test here

If the pyrroles are not isolated and instead are measured together with other chemicals like urobilinogen, then it is not a useful clinical test.

It is UNEQUIVOCAL that kryptopyrrole is NOT what is being measured.

“HPL” is an artefact of the testing process and not the product which is being tested (although, its molecular weight is used in calculations as it the most related reasonably stable common intermediate in the oxidative degradation of regulatory haem components.

[If a patient fails to respond to treatment, then the test has been conducted incorrectly, the diagnosis is incorrect or treatment plan is inadequate. “[intro to medicinal diagnostics 1A]”

The assay reagent (p-N,N-dimethylaminobenzaldehyde) also reacts Regulatory haem components such as bilirubin and urobilinogen causing interference (the interference of bilirubin is minimal due to the spectroscopic characteristics of the bilirubin-DMAB compound). Bilirubin is a bile component and urobilinogen is the result of gut microflora action on bile/bilirubin. These compounds are not directly related to oxidative stress, rather they are related to adrenal function and HP axis communication.

A separate measure of Urobilinogen can offer differential diagnostic ability to ascertain other underlying issues which may present as mental distress and digestive system dysfunction i.e. in cases of blocked bile flow, anxiety and digestive dysfunction (diarrhea) may be present.

A high measure of pyrroles in urine, when combined with clinical presentation, can give a patient hope that the problem can be managed, resolved and a return to normal health is possible. A practitioner who does not use a test that provides distinguishable measurements of the pyrrole fraction and urobilinogen is not serving their clients well.
Publication will occur in due course.

What’s Included In Our Pyrrole Test?

Pyrrole Test, Applied Analytics Laboratories

Pyrrole Test, Applied Analytics Laboratories
This simple test requires you to provide a urine sample. The DMAB active pyrroles are a waste product not found in healthy urine and indicates that your body is under stress. Co-testing of Zinc, copper and other biomarkers informs your practitioner what you need to repair your biochemistry and reduce your symptoms. The test also measures a by-product from gut function called Urobilinogen.

Urine sample collection can occur at your practitioner’s rooms if they have been approved by AAL.

Most collection occurs at a collection agency. See below for your nearest agency or ask your practitioners office. If the sample is not collected, handled correctly, or doesn’t arrive at the laboratory in appropriate condition, AAL will order a recollect. Click Here

Under normal circumstances, AAL usually tests the urine sample within in seven working days. The results are uploaded, faxed, phoned, or emailed to your practitioner.

Your practitioner will receive:
– Measure of Pyrrole
– Measure of Urobilinogen
– Specific Gravity
– Siemens 10SG Point of Care urinalysis.
– Results, report and after service consultation.

Your first Pyrrole test should be before commencing treatment. Re-testing is required after a few months to evaluate your response to treatment and to then review your management plan.

AAL test sample preparation protocols have not changed since commencing operation. The results are now refined to account for the major interference(s) with the pyrrole measure. A separate measure of Urobilinogen has provided a far more valuable test for the practitioner as it provides much more information. Using the correct two factor analysis means that AAL has largely eliminated the generation of false positive and false negative results
The AAL urinary pyrrole test is the most accurate urinary pyrrole test available.

Samples that display unusual characteristics are explored to confirm the source of the unusual event. Under NPAAC guidelines the practitioner is alerted to results that lay outside the testing range.

This follows on from the Pfeiffer/Walsh team’s proposal that pyrroles are a measure of oxidative stress. Due to equipment limitations, they did not have the technology to prove this. AAL have proven that their original premise of this test applies.

To reiterate, Elevated urinary pyrroles is not a condition, syndrome, disorder, genetic or an auto immune disease. It tells your practitioner if you have high levels of oxidative stress which must be managed.
Re-testing also shows practitioners the response to treatment.

NOTE WELL:
Pyrrole is only found in urine. Any practitioner who asks you to do any other test to measure pyrrole does not understand the biochemistry.

What is Urobilinogen?

In 2018, an Australian laboratory (Applied Analytical Laboratories) became the first to include measurement of urobilinogen in their testing process.

Urobilinogen is a by-product of normal gut function as opposed to oxidative stress and is co-measured in the assay.
Urobilinogen is much more stable than the pyrrole fraction and does not breakdown as rapidly in light and heat (although, it still does). Hence failure to follow the strict collection, storage and transport protocols results in the break-down of the Pyrrole fraction of the urine and only the Urobilinogen portion is measured.

Separation of these components allows for identification of additional pathologies (e.g. obstructed bile duct, haemolytic anaemia, or hepatitis) and is critical for correct interpretation. The distinction allows a more accurate identification of the composition of the sample as falsely treated results can exacerbate symptoms and/or lead to vitamin B6 toxicity.

The AAL laboratory report gives clear reference ranges for both Urobilinogen and Pyrroles.

Pyrrole Test, Applied Analytics Laboratories
Pyrrole Test, Applied Analytics Laboratories
The DMAB active which is testing as pyrrole is mostly UROBILINOGEN. Sullivan and Nicolaides, in 2016, conducted in house investigation and found this so and ceased collection of urine for Pyrrole testing. Since 2018 AAL have distinguished Pyrrole and Urobilinogen in the assay and given measures for both.

Interference studies show Urobilinogen interferes greatly with the pyrrole reading (even at the “recommended” second void specimen), thus adjustment of collection times does not adequately account for this. The use of the second morning void historically been chosen due to a minimum of urobilinogen at this time however, urobilinogen levels at this time can still be greater than the pyrrole fraction. This is not to say the use of this method has been of no value. The demonstration that there is a product in the urine which has some association with Mental Health has given patients hope and directed patient care. AAL have studied the degradation of both the Urobilinogen and the Pyrrole fraction and have confirmed Pyrrole is the result of oxidative stress (as taught by Bill Walsh), not the cause of ”pyrroluria” as it has commonly become confused. In comparison, the Urobilinogen is relatively stable, while the pyrrole fraction degrades rapidly. This demonstrates the need for careful collection, transport, and storage, as per the AAL SOP 3001 Urinary Pyrrole Protocol.

Benefits of a separate Urobilinogen measure

AAL now screen all samples using an automated Siemens 10SG system (and AAL participate in RCPA run proficiency testing for this system) This also enables AAL to continue to provide a fixed stable reference assay, ties our assay in with current conventional haematology and provides an extra level of assurance to aid diagnosis with the additional detection of bilirubin, blood, leukocytes, Nitrites, Protein, Ketones, and Glucose. The 10SG urobilinogen also offers a confirmation of the AAL measurement at urobilinogen levels >16µmol/L.

Provided samples are collected correctly the absence of Urobilinogen can also indicate biliary obstruction, for which AAL has a distinct profile.

The providing of a Urobilinogen measure as a possible differential diagnosis of mental health issues provides a level of care not previously possible.

WHAT IS PYRROLES IN URINE TEST?

Pyrrole Test, Applied Analytics Laboratories
Pyrrole Test, Applied Analytics Laboratories

To be scientific:
The test is a co-measure of TWO different processes.

1. Circulating oxidative stress load on regulatory haem
2. Result of bacterial action in the gut on bile—generating urobilinogen. Urobilinogen’s measurement can indicate the statis of biliary flow. I.e., The absence of urobilinogen can indicate biliary obstruction, and its presence in elevated quantity can indicate renal dysfunction (the kidneys perform conversion of urobilinogen to urobilin. High levels (>16µmol/L) can indicate the impairment of this conversion).

The test is only useful if collection, storage, and transport protocols are followed.

In non-medical terms:

1. Urinary Pyrroles
– Urinary pyrroles are a waste product not normally found in the Urine of healthy people.
– DMAB active pyrroles in urine tell us the body is under stress.
– The amount of Pyrrole tells us how much stress.
– The change in the amount of pyrrole tells us how the body is responding to treatment.
– Lowering pyrrole levels can result in improving the biochemistry.

2. Urobilinogen
– Is found in small amounts in the urine of healthy people.
– Too much Urobilinogen can be related to poor Kidney function.
– Too little can indicate blocked bile ducts (commonly caused by gallstones). Some medications (such as statin/cholesterol lowering drugs) can reduce biliary flow.
– Both levels of Urobilinogen can present with mental health like symptoms.

History

Bio Balance Health introduced the pyrroles in urine test into Australia.

After using the test for a few years, we asked the question:

What is being tested?
Pyrroles in urine are tested to examine its relationship to stress. Research has also provided a separate measure for urobilinogen. Both pyrroles and urobilinogen are co-measured in the assay with similar coloured mauve complexes. Advanced sample preparation and spectroscopic measurement are required to differentiate the components.
Urobilinogen is relatively stable and does not breakdown in light and heat as rapidly as the pyrrole fraction does.
(Pyrroles in urine are waste product generated by oxidative fragmentation of regulatory haem and are extremely labile (rapidly broken down by heat and light.))

The above statements are a result of collaborative research efforts between AAL and Bio Balance Health, as well as Griffith and Adelaide Universities.

We can now say that any previous publication or research or opinions or social media sites are all based on incorrect and inaccurate measurements (and with the incorrect premise that “pyroluria” is a diagnosis).

Through our research this test is now very valid as it now provides excellent guidance:
• regarding severity of oxidative stress.
and
• different diagnoses which should be investigated.

It is essential doctors understand these differences as incorrect use of medication and/or nutrients may increase the patient’s symptoms.

BBH trains doctors to use these skills. We work closely with BBH.
AAL also works closely with Bio Balance Health Ltd to conduct research.
The most recent collaboration has begun between AAL and QUT Centre for Genomic and Personalised Health examining treatment resistant depression. https://www.biobalance.org.au/

Much of our work is pre-publication.

To ensure we can correct much of the misinformation which is stated as “fact” about pyrroles in urine, we have produced a lecture detailing the science and the clinical applications of this test.

The lecture “The Science of Pyrroles in Urine” is available to medical professionals and the public.

All profits go to further fund our original research.
Pyrrole lecture…… https://biobalance.org.au/events/

Our research is self-funded. If you wish to increase and/or speed up our projects please donate. https://biobalance.org.au/make-a-donation/

Note. Not all mental health is associated with a high pyrrole measure. For those with high pyrroles it is very comforting to know there is a reason for feeling so messy and that something practical can be done to help.

Pyrrole, Kryptopyrrole, Pyrrole Disorder, Pyrrole Condition, Pyrroluria, Kryptopyrole, Pyroluria, Pyrrole Syndrome.

These are all names used to describe a chemical found in the urine which has been clinically shown to be associated with several chronic mental health issues. It has also historically been referred to as the MAUVE FACTOR.

No research has been conducted to show it is a condition, syndrome, or disorder, or genetic in origin.

Our research has shown:
A measure of pyrroles in urine is not a diagnosis, but a biomarker of oxidative stress.
For treatment, the stress must be managed for long term reduction of symptoms, along with re-balancing of biochemistry.
The correct term is: High or Elevated Urinary Pyrroles.

Urinary Pyrroles are unstable; thus it is essential that collection and transportation of urine is done under strict standardized conditions to maximize reliability of results. Poor collection and handling can result in false negative readings, or false positive.

No research has been conducted to show it is a condition, syndrome or disorder, autoimmune disease, or genetic.

To repeat. It is not a diagnosis, but a measure of oxidative stress which must be managed for long-term reduction of symptoms.

Work has also shown, as pyrroles in urine are a waste product, therefore unstable, it is essential that collection and transportation of urine is done under strictly standardized conditions to maximize the reliability of results. Poor collection and handling can result in false readings and poor patient outcomes.

What is important is the focus on the urine measure as a biomarker of oxidative stress which directs patient care to identify and manage the underlying oxidative stress. A doctor trained by Bio Balance Health will know how to do this and manage the process which is individualised to each patient.

The identification of High Urinary Pyrroles has changed many lives as reduction of the pyrrole measure commonly correlates with reduction of symptoms.

The understanding that very High Urinary Pyrroles is also often associated with some forms of mental ill-health has reduced the stigma and the loss of hope felt by many with a mental health diagnosis.

The process to reduce high pyrroles enhances current chronic health management.

Symptoms associated with high pyrrole vary as the underlying stress varies. This explains why there are so many symptoms and how each person with high urinary pyrroles manifests different responses. It is important your practitioner lists all your symptoms so which symptoms show treatment response can be observed. High Urinary Pyrroles are only found in certain types of mental health.

In 2018 Applied Analytical Laboratories became the first to have selectivity in their assay which enables the distinction between chemical species pyrrole from urobilinogen, and provides separate measures for both, as well as the CORRECT scientific measure of Specific Gravity (SG).

Urobilinogen is a by-product of normal gut function as opposed to oxidative stress and is co-measured in the assay.
Separation of these urine components allows for identification of additional pathologies (e.g. obstructed bile duct, haemolytic anaemia, or hepatitis) and is critical for correct interpretation.

The distinction allows a more accurate identification of the composition of the sample as treatment plans based on false results can exacerbate symptoms and/or lead to vitamin B6 toxicity.

The AAL Laboratory report will give clear reference ranges.

AAL recommends you use Bio Balance Health trained doctors who have experience in managing the process. Self-diagnosis and self-medication are fraught with difficulties and will not achieve optimal results.

Applied Analytical Laboratories Pty Ltd (AAL) is the only dedicated Pyrrole testing laboratory in Australia. AAL are the leaders in investigating the origin of elevated urinary pyrroles, both in their own processes and quality control, and through active research collaboration with university researchers.

SOME MORE SCIENCE

For those who want a bit more science.

The Pyrrole moieties that are measured are labile (reactive) intermediate fragments produced by the attack of reactive oxygen species on regulatory haem. This system has been suggested as a” last resort” anti-oxidant system of the body (bilirubin, for example, is a powerful anti-oxidant yet is physiologically toxic (we have scientific references for this).
The coordination, or reaction, between pyridoxine/pyridoxal and any pyrroles, or indoles, does NOT occur (if there was reaction of this nature all muscle contraction of every species on earth would not occur as serotonin/5HT is an indole).

No research has been performed or published to show Pyrrole is genetic or a disorder.

Conversely, we suggest that it is a condition manifested by excess oxidative stress.

Kryptopyrrole is 3-ethyl-2,4-dimethyl pyrrole and is used as one of the calibration references used for AAL’s assay.

Kryptopyrrole is NOT the active measured in the assay. It also has found successful application in our laboratory as a research chemical (where it has been used to produce HPL (the pyrroline-one form) and associated analogues for study. This work has made major inroads into elucidation of the mechanisms of the free radical oxidation (oxidation with reactive oxygen species).

Hydroxyhemepyrrolenone (aka HPL) is generated as an artifact during the testing process and is not the actual form of the analyte measured. HPL is also unstable (and rapidly disassociates) under the typically employed mildly acidic chromatographic conditions (which is why it is also difficult to measure accurately with current HPLC-MS technology).

Urobilinogen is a by-product of normal gut function, and levels can fluctuate greatly from morning to evening under normal circumstances, or times of adrenal stimulation/relaxation (through anxiety). It is not directly related to oxidative stress.

Separate measure of Urobilinogen can demonstrate other underlying issues which may present as mental distress, for example, biliary obstruction or renal failure. This makes this test very useful as an insight to other pathologies.

What do I do if I have High Urinary Pyrroles?

It is important to primarily identify and manage the cause of the underlying oxidative stress.

Measures of biochemistry are essential to direct re-balancing of nutrients lost because of the excess demand caused by a stress reaction.

A doctor trained by Bio Balance Health in the Walsh/Pfeiffer method will know how to do this and manage the process which is individualised to each patient.

Exploration of a product in urine associated with mental health started with Abram Hoffer and Carl Pfeiffer (physician and biochemist) when they explored further the observation that extracts of urine from schizophrenics turned mauve upon the addition of a specific reagent.

The identification of High Urinary Pyrroles has changed many lives as reduction of the pyrrole measure correlates with reduction of symptoms of many unexplained health issues.

The understanding that very High Urinary Pyrrole is also often associated with some forms of mental ill health has reduced the stigma and the loss of hope felt by many with a mental health diagnosis.

The processes to reduce high pyrroles are not meant to replace current chronic health management but to enhance it.

Symptoms associated with high pyrrole vary as the underlying stress varies. This explains why there are so many symptoms listed as associated with high pyrroles and how each person with high pyrroles manifest different responses.

It is apparent to Bio Balance Health doctors that management of sustained stress, as indicated by high pyrroles in urine measure, and balancing of biochemistry as indicated by related pathology testing, has been life changing for many people.

Bio Balance Health supports further research in this area.

Brett Lambert, principal of AAL, is excited to be leading this research.

Urine Collection Process for Pyrrole Analysis
“Where do I get a Urine Pyrrole Test ?”

Pyrrole Test, Applied Analytics Laboratories

Urinary Pyrrole Analysis
Request/Payment Form

Step 1

Go to a Doctor who is Bio Balance Health trained and has done the Science of Pyrroles in Urine lecture.

If your Doctor has provided you with an Applied Analytical Laboratories Urinary Pyrrole Test request form, click to find your nearest collection agency.

If you are a new practitioner looking to refer a patient, please email to request an AAL Urinary Pyrrole Test request form.

Step 2

Get a Urinary Pyrrole Test Request Form

If your Doctor has provided you with an Applied Analytical Laboratories Urinary Pyrrole Test request form, click to find your nearest collection agency.

If you are a new practitioner looking to refer a patient, please email to request an AAL Urinary Pyrrole Test request form.

Step 3

Cease Supplements?

First time test? Patients must have not started zinc supplements or cease supplements 72 hours prior to tests (to establish a base-line).

Follow up test? Do NOT cease Zinc if it is a follow-up test.

Step 4

Step 3. Note: Medications

Medications used with Rheumatoid Arthritis and/and Urinary Tract Infections can interfere with the test, as does Rifampicin which changes the urine to a red/mauve colour. If you are taking these medications consult your physician as you will need to complete the course and/or abstain for a wash out period of 4 weeks prior to collecting a sample. For some, this is not possible.

Step 5

Time of Collection?

Any time of the day is possible. Early in the day is preferable.

It is important that the time of collection needs to be noted on the form in order to help account for obtained Urobilinogen levels. Very high levels of urinary bilirubin DO also interfere. However this is rare and you will be informed if this is the case.

Blood does NOT affect the test result, so spotting at the beginning or end of menstrual cycle will not interfere with the result. It is NOT recommended to collect during peak menstruation.

Step 6

Use Applied Analytical Specimen Jar

Urine is to be collected only in the AAL specimen jar containing medical grade preservative and immediately covered with foil which protects it from light and heat and from incidental high energy electromagnetic radiation (EMR). Any other jar will not have the required preservative inside the jar.

Step 7

Flash Freeze

The sample is to be frozen immediately to -30’C and kept covered and frozen from the time of collection, during transport, and until the time of testing in the laboratory.

Step 8

Transport (Collection Agency)

Samples are transported in dry ice to keep the relative temperature below freezing (at -30oC). Your results will be sent back to your doctor.

Tips

Pyrrole Results very high or very low?

According to NPAAC guidelines, we will notify the practitioner when the urinary pyrrole result is greater than 400ug/dL, urobilinogen is greater than 16µmol/L, or other parameters are detected (such as, bilirubin, blood, ketones, protein, nitrites and leucocytes). These can illuminate if a patient has liver/kidney function issues, diabetes, infection or other protein dysregulation factors. Applied Analytical Laboratories (AAL) is the only facility to meet this requirement and successfully participate in RCPA conducted proficiency programs for this system as an extra level of assurance. 

YOUR DOCTOR WILL BE NOTIFIED IMMEDIATELY OF RESULTS NEEDING URGENT ATTENTION.

Payment Process

The Urinary Pyrrole Request Form the doctor gives you to take to the collection agency will have a section to fill out your credit card details for the test. The test cannot occur until this is filled out.

Your collection agency will charge you separately for collection and handling fee.

Some practitioners in the Brisbane and Gold Coast area have an arrangement where the sample is collected on behalf of AAL and this reduced handling fee is included in their request form.