as a Metabolic Disorder.
analytical methods for detecting, and to some extent determining, these
compounds in the urine are available today.
One of the most popular methods is HPLC (High Performance Liquid
Chromatography). HPLC uses a liquid mobile phase to separate the
compounds of a mixture. Mixtures (samples) are first dissolved in a
solvent, and then forced to flow through a chromatographic column under
high pressure. Inside the column, the mixture is separated into its
components, as it passes [elutes] down the column. Positioned
immediately after the column is a detector, which emits a response as a
function of the components of the sample passing through it. There are
several types of detector used for this purpose; the one currently used
in this analysis is a ultra-violet (UV) detector. A UV detector measures
the ability of a sample to absorb UV light. The majority of organic
compounds can be analysed using this kind of detector. Depending on time
taken for the compound to elute through the column, and the ability of a
compound to absorb UV radiation, we are provided with data about the
type of compound being detected and a general idea of how much of the
compound is present (see published
references for HPLC protocols).
qualitative results and interpretations of example urine profiles can be
viewed in the following articles: Autism
as a Metabolic Disorder and Urinary
Profiles from people with Autism (apologies for the poor quality of
the results shown). The
control urine sample spiked with casomorphin peptides (215nm).
for urinary profiling
normal timescale of events means that participants’ (or participant
carers) complete and return the questionnaire back to us at the Unit
before any sample is submitted. Please note all details taken from the
questionnaire are subject to the strictest confidence, and are entered
on to our research database (access to the database is by authorised
personnel only). This has been ratified by the
for the collection of a urine sample for HPLC analysis (v3.0)
Research Unit (ARU), School of Health, Natural & Social Sciences,
University of Sunderland,
The transport of organic / biochemical products by the postal service
demands that strict health and safety procedures are followed regarding
sample packaging and clear labelling of goods carried. It is your
responsibility to ensure that the sample is packaged correctly and
returned to the ARU in a non-hazardous condition. Please read
through the following instructions for safe management and transport of
30ml tube (enclosed in the protective plastic postal container) you have
been provided with will contain a small quantity of a preservative* (thymol).
preservative may not be visible with the naked eye, although does have a
approximately 20ml of urine from the first production urine sample in
the morning, mid-stream (if possible**). Do not exceed the 20ml
mark on the tube or the sample tube may burst whilst frozen. Make
sure the cap on the tube is firmly closed, and the name and date of
birth of the person from whom the sample was taken is written on the
soon as the sample is taken, place the tube in a freezer for at least 24
the frozen sample tube in the protective plastic container provided.
At this point we also recommend that the protective container be
securely fastened with an elastic band*** to prevent opening during
transit. Fill out the form shown at the bottom of this sheet.
the protective container in a padded envelope (a jiffy bag is ideal) and
mail first-class post to us at the address given above, clearly marking
the package as containing a MEDICAL SAMPLE FOR ANALYSIS. Mailing
is best done at the beginning of the week to allow for receipt of the
delivery. Please note the ARU is not open to receive samples at
weekends or bank holidays.
are performed on a batch system with a current waiting time of 1 week
(subject to alteration). A
copy of the results will be sent to both parents
\ carers and the patient’s GP.
do not send a sample unless you have completed and returned the
of urinary analysis
October 2003: Due
to an upgrade of technical equipment, the ARU urine profiling service is
now able to offer more confirmatory power regarding the analysis of
urine samples. We are now using a HP1100
series system with UV-DAD. New protocols for presentation of results
have been implemented. Further
examples of the type of results obtained from our method are shown here
(please note this is a Microsoft Word document v2000).
information sheet is designed to give a general overview of the urinary
results obtained using HPLC analysis (High Performance Liquid
Chromatography) as part of the research carried out at the Autism
Research Unit at the
article has recently appeared in the scientific literature regarding the
relevance of IAG to autism spectrum disorder (Wright et
al, 2005*). The article
reports that quantitative levels of urinary IAG, although elevated in
autism compared to controls, were not significantly different from urine
samples taken from control (non-autistic) participants.
These findings are in contrast to those we have previously
reported in the scientific literature regarding quantitative levels of
IAG (Bull et al, 2002) [see
a list of publications by
the Autism Research Unit].
these recent findings are important, they do not impact significantly on
the method of analysis used to produce the current results.
This is based on a qualitative method (not specifically designed
to measure the amount of IAG present in the urine sample) to ascertain
the appearance of IAG relative to other compounds in the important
biological area (18-30 minutes on graph a).
This method showed significant differences in the presence of IAG
in autism samples compared to non-autism and learning disability
controls (Alcorn et al,
the issue of a possible relationship between urinary IAG and the use of
dietary intervention, this recent paper does not provide any further
details in addition to what we have previously published in the
scientific literature (Whiteley et
a precautionary measure, we feel we must reiterate the following points:
Our methodology is still experimental.
Any decision to act upon these findings must reside entirely with
the parents and the person with autism.
This test does not constitute a diagnosis or confirmation of a
diagnosis of autism or associated spectrum disorder.
We suggest the agreement of the physician involved with each
individual case be obtained before commencement of dietary intervention.
The involvement of a registered dietitian / nutritionist is
recommended in the production and implementation of any dietary
Wright B. et al. (2005) Is the presence of urinary
indolyl-3-acryloylglycine associated with autism spectrum disorder? Dev
Med Child Neurol. 47 (3): 190-192
Home Page - http://osiris.sunderland.ac.uk/autism/index.html
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Metabolic problems show up first as blood sugar abnormalities, &/or early on
Repost of http://osiris.sunderland.ac.uk/autism/hplc.htm