Characterization of Ankylosing Spondylitis and Rheumatoid Arthritis Using 1H NMR-Based

Characterization of ankylosing spondylitis and rheumatoid arthritis using 1H NMR-based metabolomics of human fecal extracts

Shao Tie-juan+, He Zhi-xing+, Xie Zhi-jun, Li Hai-chang, Wang Mei-jiao, Wen Cheng-ping*

College of Basic Medical Science, Zhejiang Chinese Medical University, Hangzhou 310053, China

Corresponding author. Address: College of Basic Medical Science, Zhejiang Chinese Medical University, Hangzhou 310053, China.

“+”: these authors contributed equally to this work.

Tel.: +86-57186613587.

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Summary of supporting information:

A.SUPPORTING FIGURES S1

Figure. S1 The detailed concentration of 46 identified metabolites between healthy controls and AS, RA patients. The concentration of metabolites between control and AS/RA were analyzed by no-parametric test.

1. SUPPORTING TABLES S1-S2

Table S1. List of the metabolites found in the contents of fecal extracts.

Table S2. The Pearson correlation coefficient between metabolites and CRP, ESR, Age or Sex.

Figure. S1 The detailed concentration of 46 identified metabolites between healthy controls and AS, RA patients. The concentration of metabolites between control and AS/RA were analyzed by no-parametric test.

Table S1. List of the metabolites found in the contents of extracts of feces a

N. / Metabolites / Moieties / δ 1H (ppm) and multiplicity
1 / butyrate / CH3,βCH2,αCH2 / 0.90(t),1.56(m),2.15(t)
2 / α-ketoisocaproate / 2*CH3,CH,CH2 / 0.92(d),2.06(m),2.61(d)
3 / isoleucine / δCH3,γCH3,γCH2,γ'CH2,βCH,αCH / 0.94(t),1.01(d),1.25(m),1.48(m),1.98(m),3.67(d)
4 / leucine / δCH3,δCH3,γCH,βCH2,αCH / 0.96(d),0.97(d),1.69(m),1.71(m),3.74(t)
5 / valine / γCH3,γCH3,βCH,αCH / 0.99(d),1.04(d),2.27(m),3.62(d)
6 / propionate / CH3,CH2 / 1.06(t),2.19(q)
7 / ethanol / CH3, CH2 / 1.19(t),3.67(q)
8 / lactate / βCH3,αCH / 1.33(d),4.11(q)
9 / threonine / γCH3,αCH,βCH / 1.33(d),3.59(d),4.26(m)
10 / alanine / βCH3,αCH / 1.48(d),3.79(q)
11 / lysine / γCH2,δCH2,βCH2,εCH2,αCH / 1.48(m),1.72(m),1.90(m),3.03(t),3.76(t)
12 / citrulline / γCH2,βCH2,δCH2,αCH / 1.57(m),1.87(m),3.15(t),3.75(t)
13 / arginine / γCH2,βCH2,δCH2,αCH / 1.73(m),1.93(m),3.03(t),3.75(t)
14 / acetate / CH3 / 1.92(s)
15 / proline / γCH2,βCH2,β'CH2,δCH2,δ'CH2,αCH / 2.01(m),2.07(m),2.36(m),3.34(m),3.45(m),4.13(m)
16 / methionine / δCH3,βCH2,γCH2,αCH / 2.14(s),2.16(m),2.65(m),3.86(m)
17 / glutamate / βCH2,β'CH2,γCH2,αCH / 2.10(m), 2.09(m),2.36(m),3.77(m)
18 / succinate / 2*CH2 / 2.41(s)
19 / glutamine / βCH2,γCH2,αCH / 2.10(m),2.46(m),3.77(m)
20 / aspartate / βCH2,β'CH2,αCH / 2.68(m),2.82(m),3.91(m)
21 / asparagine / βCH2,β'CH2,αCH / 2.86(dd),2.96(dd),4.00(m)
22 / trimethylamine / 3*CH3 / 2.88(s)
23 / dimethylglycine / CH3 / 2.76(s)
24 / creatine / CH3,CH2 / 3.04(s),3.93(s)
25 / choline / N(CH3)3,NCH2,OCH2 / 3.21(s),3.52(m),4.07(m)
26 / taurine / CH2SO3,NCH2 / 3.25(t),3.43(t)
27 / methanol / CH3 / 3.37(s)
28 / glycine / CH2 / 3.57(s)
29 / β-glucose / 2CH,4CH,5CH,3CH,CH2C6,1CH / 3.28(dd),3.41(t),3.47(dd),3.50(t),3.91(dd),4.65(d)
30 / α-glucose / 2CH,4CH,3CH,CH2C6,CH2C6',5CH,1CH / 3.53(dd),3.42(t),3.71(t),3.74(m),3.84(m), 3.84(dd),5.23(d)
31 / UDP-glucose / CH / 5.62(m)
32 / uracil / CH,CH / 5.81(d),7.54(d)
33 / fumarate / 2*CH / 6.53(s)
34 / tyrosine / βCH2,β'CH2,αCH,3 or 5CH,2 or 6CH / 3.06(dd),3.20(dd),3.94(dd),6.91(d),7.20(d)
35 / histidine / βCH2,β'CH2,αCH,5CH,3CH / 3.14(dd),3.25(dd),3.99(dd),7.08(s),7.83(s)
36 / phenylalanine / βCH2,β'CH2,αCH,2 or 6CH, 4CH,3 or 5CH / 3.13(dd),3.29(dd),3.98(dd),7.33(m),7.38(m),7.43(m)
37 / tryptophan / βCH2,β'CH2,αCH,5CH,6CH,
2CH,7CH,4CH / 3.31(dd),3.49(dd),4.06(dd),7.21(t),7.29(t), 7.33(s),7.55(d),7.74(d),
38 / urocanate / CHCOOH,CH(ring),5CH,3CH / 6.40(d),7.31(d),7.43(s),7.89(s)
39 / hypoxanthine / CH, CH / 8.20(s), 8.21(s)
40 / formate / CH / 8.45(s)
41 / inosine / CH2,'CH2,5H',4H', 2H', 8H,2H / 3.85(dd),3.92(dd),4.28(q),4.44(t),6.10(d),
8.24(s),8.34(s)
42 / bile acids / C18 axial methyl / 0.70(m)
43 / -xylose / 1CH,2CH,3CH / 4.58(d),3.24(dd),3.47(t)
44 / -arabinose / 1CH,2CH,3CH / 4.52(d),3.52(dd),3.69(m)
45 / -arabinose / 1CH,2CH / 5.21(d),3.87(d)
46 / -xylose / 1CH,2CH,3CH / 5.20(d),3.53(dd),3.68(m)

as, singlet; d, double; t, triplet; q, quartet; m, multiplet; dd, double of doubles.

Table S2. The Pearson correlation coefficient between metabolites and CRP, ESR, Age or Sex

Pearson Correlation / ESR / CRP
r / p / r / p
butyrate / -0.093 / 0.334 / -0.133 / 0.173
-ketoisocaproate / 0.253 / 0.008 / 0.034 / 0.728
isoleucine / 0.103 / 0.288 / 0.191 / 0.049
leucine / -0.079 / 0.418 / -0.156 / 0.109
valine / -0.085 / 0.379 / -0.096 / 0.325
propionate / 0.004 / 0.971 / -0.125 / 0.198
ethanol / -0.019 / 0.847 / 0.056 / 0.567
lactate / -0.099 / 0.304 / 0.000 / 0.966
threonine / 0.169 / 0.080 / 0.115 / 0.240
alanine / 0.016 / 0.871 / 0.123 / 0.203
lysine / 0.067 / 0.489 / 0.178 / 0.066
citrulline / 0.084 / 0.385 / -0.108 / 0.268
arginine / -0.028 / 0.774 / 0.026 / 0.789
acetate / 0.067 / 0.491 / 0.043 / 0.661
proline / 0.190 / 0.048 / 0.031 / 0.750
methionine / -0.145 / 0.134 / -0.096 / 0.327
glutamate / -0.035 / 0.719 / 0.079 / 0.416
succinate / 0.010 / 0.917 / -0.016 / 0.869
glutamine / 0.102 / 0.293 / 0.075 / 0.440
aspartate / 0.123 / 0.203 / -0.025 / 0.797
asparagine / 0.208 / 0.030 / 0.045 / 0.646
trimethylamine / 0.153 / 0.113 / 0.015 / 0.875
dimethylglycine / 0.119 / 0.217 / 0.046 / 0.639
creatine / -0.089 / 0.357 / 0.035 / 0.720
choline / 0.086 / 0.374 / 0.193 / 0.046
taurine / 0.080 / 0.411 / 0.166 / 0.087
methanol / 0.136 / 0.160 / 0.207 / 0.032
glycine / 0.052 / 0.594 / 0.076 / 0.434
-glucose / 0.020 / 0.838 / 0.053 / 0.582
-glucose / -0.034 / 0.727 / 0.060 / 0.538
UDP-glucose / -0.005 / 0.963 / -0.193 / 0.046
uracil / 0.016 / 0.869 / -0.015 / 0.882
fumarate / 0.201 / 0.036 / 0.152 / 0.117
tyrosine / -0.069 / 0.478 / 0.081 / 0.405
histidine / 0.165 / 0.087 / 0.092 / 0.346
phenylalanine / -0.023 / 0.809 / -0.045 / 0.644
tryptophan / 0.000 / 0.999 / 0.251 / 0.009
urocanate / 0.115 / 0.234 / 0.035 / 0.720
hypoxanthine / -0.097 / 0.318 / -0.174 / 0.074
formate / 0.023 / 0.808 / 0.096 / 0.323
inosine / 0.192 / 0.045 / 0.176 / 0.070
bile acids / -0.072 / 0.454 / -0.034 / 0.730
b-xylose / 0.010 / 0.918 / -0.057 / 0.556
-arabinose / 0.128 / 0.184 / 0.004 / 0.964
-arabinose / -0.075 / 0.438 / -0.103 / 0.293
xylose / 0.031 / 0.752 / -0.124 / 0.204