Urine analysis in relation to diet and ASC

Transcription

Urine analysis in relation to diet and
ASC
Kevin Carr
Director
ESPA Research
Where research leads we will follow
ESPA Research Limited
Company Registration: 6862992
The Robert Luff Laboratory,
Unit 133I Business & Innovation Centre (BIC)
Sunderland Enterprise Park, Wearfield, Sunderland SR5 2TA, UK
Website: www.espa-research.org.uk
Telephone: 0191 549 9300
Email: [email protected]
A not-for-profit subsidiary of ESPA
ESPA Registered charity: 1037868
How does urine analysis help with diet
•The compounds in urine all come from the things we eat
•Some directly
•Amino acids, peptides etc
•Some indirectly
•Compounds produced by gut bacteria
Telephone: 0191 549 9300
Different gut bacteria and different diets
Oats
Wheat
Rice
Vessel 1
Vessel 2
Vessel 3
IAG Story
• HPLC showed a peak which was higher in some ASC
samples, compared with controls
• Peak identified as indolylacryloylglycine (IAG)
• IAG known to be connected with PKU and Hartnup Disease
• IAG known to be a metabolite of tryptophan
• Where IAG is produced is not known
Telephone: 0191 549 9300
3-Indole Acetic Acid (3-IAA)
3-Indole Propionic Acid
3-Indole Pyruvic
Acid
TRYPTOPHAN
3-Indole Acrylic Acid
(IAcrA)
Indole and Indican
Kynurenine and
derivatives
(tryptophan hydroxylase)
5-Hydroxy Tryptophan
(5-Hydroxytryptophan decarboxylase)
Serotonin
3-Indolyl Acryloyl Glycine
(IAG)
5-Hydroxy Indole Acetic Acid (5-HIAA)
Melatonin
Bufotenin
Telephone: 0191 549 9300
Why analyse urine
• Blood and plasma too invasive a technique
• Urine contains the majority of metabolites
excreted from the body and is collected non
invasively
Telephone: 0191 549 9300
Problems with the analysis of urine
• Very complex mixture of compounds
• Metabolites of interest present in very small
quantities
• Metabolites can change due to the presence of
other chemicals
Telephone: 0191 549 9300
x103 DAD1-A:Sig=280,4Ref=460,1001550.d
2.316
2.25
2
4.546
1.761
1.75
6.863
1.5
1.25
1
3.535
4.964
0.75
0.5
7.383 8.393
6.309
5.885
9.703
7.740
25.276
11.414
11.874
0.25
12.693
14.766
13.544
15.853 16.892
18.200
19.628 20.547
0
1
2
3
4
5
6
7
8
9
10
11
12
13
i=326,4Re=
f4601
,001550.d
x102 DAD1-B:Sg
14
15
16
17
Responsevs.AcquisitionTm
i e(mn
i)
3.5
3
2.5
2
1.5
1
0.5
19
20
22.217
21
22
23
24
25
26
27
28
29
30
15.447
11.403
5
4.5
4
18
24.138
2.178
6.851
1.776
9.071
3.752
5.126 6.083
7.868
9.471
10.275
12.007 12.963
14.480
15.95416.766 17.917
19.486 20.654
0
1
2
3
4
5
6
7
8
9
10
11
12
13
14 15 16 17
Responsevs.AcqusitionTm
i e(mni)
18
19
20
21
24.093 25.156
22.040
22
23
24
25
29.169
26
27
28
29
30
Telephone: 0191 549 9300
How does ESPA Research solve these
problems
• Development of new separation methods
• State-of-the-art analysing equipment and
software
• Analysts who think in strange ways
x10 7
2
Telephone: 0191 549 9300
+ESI TIC S c a n Frag=110.0V 1550.d
1
1
1.75
1.5
1.25
1
0.75
0.5
0.25
x10 4
+ESI EIC(90.0550) Scan Frag=110.0V 1550.d
4.342
1
1
8
6
4
0.771
2
0.914
2.424
x10 5
3
4.474
1.807
0
0.815
1
1
2.5
2
1.5
1
2.182
0.5
1.289
0
x10 3
6
1.597
3.229
2.854
3.978
4.309
1
1
5
4
1.124
0.859
3
3.956
4.154
2
0.980
1
0.176
1.322
0.473
1.564
5.080
2.391
0
x10 4
1.4
4.121
1
1
1.2
5.036
1
6.017
0.8
0.6
5.422
0.4
0.2
1.079
1.774
1.344
4.452
3.471
2.799
2.281
4.694
5.223
5.565
6.965
7.957
7.273
8.320
8.607
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
2.2
2.4
2.6
2.8
3
3.2
3.4
3.6
3.8
4
4.2
4.4
4.6
4.8
5
5.2
5.4
5.6
Counts vs. Acquisition Time (min)
5.8
6
6.2
6.4
6.6
6.8
7
7.2
7.4
7.6
7.8
8
8.2
8.4
8.6
8.8
9
9.2
9.4
9.6
9.8
Telephone: 0191 549 9300
x10 4 +ESI Scan (4.320-4.342 min, 3 scans) Frag=110.0V 1550.d
5.6
249.1555
263.1461
271.1645
282.1197
5.4
5.2
5
4.8
4.6
4.4
273.1669
4.2
4
3.8
3.6
3.4
268.1039
3.2
3
307.1718
2.8
343.1609
2.6
2.4
2.2
2
261.1306
1.8
293.1466
345.1874
1.6
1.4
1.2
305.1563
266.1586
319.1717
1
0.8
0.6
0.4
0.2
0
242 244 246 248 250 252 254 256 258 260 262 264 266 268 270 272 274 276 278 280 282 284 286 288 290 292 294 296 298 300 302 304 306 308 310 312 314 316 318 320 322 324 326 328 330 332 334 336 338 340 342 344 346 348 350 352
Countsvs. Mass-to-Charge (m/z)
Telephone: 0191 549 9300
x10 5 +ESI Scan (15.420-15.644 min, 20 scans) Frag=100.0V 2631-r002.d
245.0922
4.5
(M+H)+
O H
4.25
H
4
3.75
H
3.5
O
O
3.25
N
3
170.0599
H
2.75
H
2.5
H
2.25
H
2
1.75
H
1.5
1.25
H
1
511.1589
121.0696
N
H
0.75
922.0098
0.5
H
0.25
H
0
50 75 100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500 525 550 575 600 625 650 675 700 725 750 775 800 825 850 875 900 925 950
Counts vs. Mass-to-Charge (m/z)
IAG
Telephone: 0191 549 9300
Present Research
• Identify compounds which may be of relevance
• Determine ratios between urine compounds
•Some known some unknown
•Compare ratios of ASC subjects and controls.
Telephone: 0191 549 9300
Phenylpyruvate
Tetrahydrobiopterin
Valine
Homocysteine
3-(3-hydroxyphenyl)-3-hydroxypropionic acid
Leucine
2-methylbutyryl glycine
Citramalic acid
Isoleucine
Creatine
Tartaric acid (3-OH-malic)
Ornithine
Guanidinoacetate
3-oxoglutaric acid
Taurine
Bufotenine
Phenylcarboxylic acid (benzoic acid)
Methionine
5-hydroxyindoleacetic acid
Arabinose
Glycine-Serine-Glutamic acid-Asparagine
Serotonin
Hippuric acid
Salsolinol
5-hydroxyindole-3-acetic acid
4-hydroxyhippuric acid
N-methyl-Salsolinol
6-sulphatoxymelatonin
N1-methyl-2-pyridone-5-carboxamide
Norsalsolinol
Insulin-like growth factor-1
N-methyl nicotinic acid (trigonelline)
N-methyl-Norsalsolinol
Epidermal growth factor
N-methyl nicotinamide
Mono-2-ethylhexyl phthalate (MEHP)
Insulin-like growth factor binding protein-3
Dimethylamine
Mono-2-ethyl-5-oxohexylphthalate (MEOP)
Isoprostane F(2alpha)-VI
Phenylacetylglutamine
Di-2-ethylhexylphthalate (DEHP)
2,3-dinor-thromboxane B(2)
Dopamine
mono-n-butyl phthalate (MNBP)
6-keto-prostaglandin F(1alpha)
3-4 dihydroxyphenylacetic acid (DOPAC)
dibutyl phthalate (DBP)
8-hydroxy-2-deoxyguanosine (8-OHdG)
3-methoxytyramine (3MT)
MEOHP
8-isoprostane-F2alpha (8-iso-PGF2alpha)
Tyrosine
MEHHP
Xanthine
Tryptophan
Methylxanthine
Glutamine
beta-Phenylethylamine
Glutamate
3-methoxy-4-hydroxyphenyl glycol
Glycine
homovanillic acid
Phenylalanine
Glyceric acid
Asparagine
Uric acid
Histidine
Neopterin
Alanine
Biopterin
Lysine
Cysteine
ScanBrit urine samples
• Over 350 samples
• Statistical analysis between responders and non
responders
• Identification of most relevant compounds
Telephone: 0191 549 9300
What does the software do
Telephone: 0191 549 9300
Telephone: 0191 549 9300
How does urine analysis help with diet
• Discover compounds which have altered levels in ASC (and
other conditions)
• Determine the biological systems which produce these
compounds
• Establish problems in these biological systems which would
alter the levels of the compounds
• Investigate where diet/supplements may help alleviate these
problems
Telephone: 0191 549 9300
Ltd
analutos
Ltd
analutos
What is Analutos?
Why was it formed?
Ltd
analutos
What will it do?
Provide analytical services for
•Practitioners
•Parents
•Other research groups
•Industry
Ltd
analutos
How will it work?
•All samples for testing will come through Analutos
•Analutos will provide information on the test and the
results but will not give advice based on those results
•Advice on implementation of diet will still be available
from ESPA Research but parents will always be advised to
see a professional practitioner.
Ltd
analutos
Analutos – ESPA Research Link
Research findings by ESPA Research developed by
Analutos into valid tests.
Profits from Analutos donated to ESPA Research to fund
further research.
Ltd
analutos
Tests available from Analutos
•Existing IAG analysis transferred from ESPA Research
•Urine test for the quantification of all Amino Acids
•Urine test for levels of compounds known to be of
relevance in ASC (scientific literature)
•Whatever anyone wants!
Ltd
analutos
Analutos aims and objectives
•Develop tests which are relevant
•Work with practitioners and parents to develop tests
they require
•Produce results in a way that practitioners and parents
want
Ltd
analutos
Working together to find solutions
Telephone: 0191 549 9300

Urine analysis in relation to diet and ASC

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