Bile acid indices as biomarkers for liver diseases I: Diagnostic markers

Table 1 List of bile acid indices

Composition	Hepatic metabolism	Hydrophilicity	CYP8B1 activity	Intestinal contribution
Concentration of individual BA	Total sulfated	Total mono-OH	Total 12α-OH	Total primary
% of individual BA	Total G-amidated	Total di-OH	Total non-12α-OH	Total secondary
	Total T-amidated	Total tri-OH	12α-OH/non-12α-OH	Primary/secondary
	% Sulfation	% Mono-OH	CA/CDCA	% Primary
	% Amidation	% Di-OH	% 12α-OH	% Secondary
	% G-amidation	% Tri-OH	% Non-12α-OH
	% T-amidation

BA: Bile acids; G: Glycine; T: Taurine; CDCA: Chenodeoxycholic acid; CA: Cholic acid.

Table 2 Demographics

	Controls	Patients
n	103	300
Gender1
Male, female	32, 71	157, 143
Age (yr)1
mean ± SE	44.3 ± 0.64	52.1± 0.54
19-29	17	11
30-41	28	40
42-53	30	92
54-83	28	157
BMI1
mean ± SE	27.5 ± 0.28	30.9 ± 0.32
Normal BMI < 25	30	69
Overweight BMI = 25-29.9	45	104
Obese BMI ≥ 30	28	127
Race
White	88	247
Black	7	14
Asian	7	13
Hispanic	1	8
Others	0	18

¹Significant difference between controls and patients (P < 0.05).

BMI: Body mass index.

Table 3 Absolute concentrations of major bile acids in controls and patients

BA	Unamidated	G-BA	T-BA	Total
	mean ± SE, µmol/L
Controls
Unsulfated BA
LCA	0.000 ± 0.00	0.000 ± 0.00	0.000 ± 0.00	0.001 ± 0.00
UDCA	0.004 ± 0.00	0.033 ± 0.00	0.002 ± 0.00	0.038 ± 0.00
CDCA	0.003 ± 0.00	0.008 ± 0.00	0.002 ± 0.00	0.013 ± 0.00
DCA	0.022 ± 0.00	0.011 ± 0.00	0.002 ± 0.00	0.035 ± 0.00
HDCA	0.01 ± 0.00	0.00 ± 0.00	ND	0.007 ± 0.00
MDCA	0.060 ± 0.01	ND	ND	0.058 ± 0.01
CA	0.179 ± 0.03	0.067 ± 0.00	0.009 ± 0.00	0.255 ± 0.03
MCA	0.028 ± 0.00	0.287 ± 0.02	0.041 ± 0.00	0.356 ± 0.02
HCA	0.008 ± 0.00	0.016 ± 0.00	0.001 ± 0.00	0.026 ± 0.00
Other BA1	0.160 ± 0.01	-	-	0.160 ± 0.01
Total unsulfated	0.464 ± 0.04	0.422 ± 0.02	0.057 ± 0.00	0.943 ± 0.05
Sulfated BA
LCA	0.010 ± 0.00	0.780 ± 0.04	0.220 ± 0.01	1.010 ± 0.05
UDCA	0.450 ± 0.02	1.040 ± 0.05	0.030 ± 0.00	1.520 ± 0.07
CDCA	0.070 ± 0.01	2.380 ± 0.13	0.060 ± 0.00	2.510 ± 0.13
DCA	0.010 ± 0.00	2.900 ± 0.14	0.220 ± 0.02	3.130 ± 0.16
CA	0.004 ± 0.00	0.056 ± 0.01	0.126 ± 0.01	0.190 ± 0.01
Total sulfated	0.535 ± 0.03	7.170 ± 0.28	0.650 ± 0.03	8.350 ± 0.31
Overall total	1.000 ± 0.05	7.590 ± 0.29	0.710 ± 0.03	9.300 ± 0.33
Patients
Unsulfated BA
LCA	0.004 ± 0.00	0.001 ± 0.00	0.0001 ± 0.00	0.005 ± 0.00
UDCA	0.079 ± 0.03	0.410 ± 0.17	0.012 ± 0.00	0.500 ± 0.21
CDCA	0.020 ± 0.00	0.090 ± 0.01	0.100 ± 0.02	0.210 ± 0.03
DCA	0.040 ± 0.00	0.040 ± 0.00	0.010 ± 0.00	0.090 ± 0.01
HDCA	0.010 ± 0.00	0.00 ± 0.00	ND	0.010 ± 0.00
MDCA	0.050 ± 0.01	ND	ND	0.050 ± 0.01
CA	0.240 ± 0.03	0.550 ± 0.07	0.320 ± 0.08	1.120 ± 0.14
MCA	0.120 ± 0.02	1.940 ± 0.29	0.730 ± 0.09	2.790 ± 0.34
HCA	0.010 ± 0.00	0.170 ± 0.02	0.090 ± 0.02	0.270 ± 0.04
Other BA1	0.860 ± 0.13	-	-	0.860 ± 0.13
Total	0.460 ± 0.04	0.42 ± 0.02	0.06 ± 0.00	5.910 ± 0.57
Sulfated BA
LCA	0.030 ± 0.01	2.230 ± 0.20	0.650 ± 0.06	2.910 ± 0.24
UDCA	1.560 ± 0.23	15.30 ± 2.68	1.230 ± 0.27	18.10 ± 3.08
CDCA	0.190 ± 0.03	18.70 ± 1.79	1.910 ± 0.38	20.80 ± 2.07
DCA	0.040 ± 0.01	4.280 ± 0.54	0.520 ± 0.07	4.840 ± 0.58
CA	0.080 ± 0.01	0.910 ± 0.13	1.030 ± 0.21	2.010 ± 0.31
Total	1.900 ± 0.24	41.40 ± 4.12	5.340 ± 0.74	48.70 ± 4.77
Overall total	3.330 ± 0.33	44.60 ± 4.46	6.610 ± 0.85	54.60 ± 5.20

¹Other bile acids: Nor-deoxycholic acid, 12-oxo-chenodeoxycholic acid, 3-dehydrocholic acid, 6-oxo-lithocholic acid, 7-oxo-lithocholic acid, 12-oxo-lithocholic acid, isolithocholic acid, and isodeoxycholic acid.

ND: Not detected; -: Not quantified; BA: Bile acids; G: Glycine; T: Taurine; CDCA: Chenodeoxycholic acid; CA: Cholic acid; LCA: Lithocholic acid; UDCA: Ursodeoxycholic acid; DCA: Deoxycholic acid; HDCA: Hyodeoxycholic acid; MDCA: Murideoxycholic acid; MCA: Muricholic acid; HCA: Hyocholic acid.

Table 4 Representative bile acids concentrations and indices in controls vs patients

BA (µmol/L) or BA indices	Controls		Patients		Patients vs controls
	mean	SE	mean	SE	Ratio	P value
Total BA	9.30	0.33	54.6	5.20	5.87	0.000
Total LCA	1.01	0.05	2.92	0.24	2.88	0.000
Total UDCA	1.56	0.07	18.6	3.23	11.9	0.001
Total CDCA	2.52	0.13	21.0	2.09	8.35	0.000
Total DCA	3.16	0.16	4.92	0.58	1.56	0.072
Total HDCA	0.01	0.00	0.01	0.00	1.57	0.051
Total MDCA	0.06	0.01	0.05	0.01	0.90	0.992
Total CA	0.44	0.03	3.13	0.44	7.09	0.003
Total MCA	0.36	0.02	2.79	0.34	7.83	0.000
Total HCA	0.03	0.00	0.27	0.04	10.6	0.001
Other BA1	0.16	0.01	0.86	0.13	5.54	NA
% LCA	11.5	0.38	9.20	0.39	0.79	0.002
% UDCA	17.7	0.49	21.3	0.88	1.21	0.138
% CDCA	27.1	0.65	36.3	0.94	1.34	0.000
% DCA	31.1	0.68	14.6	0.53	0.47	0.000
% HDCA	0.07	0.01	0.04	0.00	0.54	0.052
% MDCA	0.64	0.04	0.36	0.05	0.56	0.135
% CA	5.25	0.27	6.27	0.25	1.19	0.064
% MCA	4.03	0.16	6.39	0.34	1.58	0.003
% HCA	0.30	0.02	0.45	0.04	1.52	0.018
Total Unamidated	1.00	0.05	3.33	0.33	3.34	0.000
Total G-amidated	7.59	0.29	44.6	4.46	5.88	0.000
Total T-amidated	0.71	0.03	6.61	0.85	9.37	0.001
% Amidation	87.7	0.47	86.9	0.65	0.99	0.053
% G-amidation	79.7	0.49	76.0	0.71	0.95	0.000
% T-amidation	7.98	0.26	10.8	0.46	1.35	0.005
Total Unsulfated	0.94	0.05	5.91	0.57	6.26	0.000
Total Sulfated	8.35	0.31	48.7	4.77	5.83	0.000
% Sulfation	88.5	0.46	82.9	0.60	0.94	0.000
Total Mono-OH	1.01	0.05	2.92	0.24	2.88	0.000
Total Di-OH	7.30	0.29	44.6	4.58	6.11	0.000
Total Tri-OH	0.82	0.04	6.19	0.65	7.52	0.000
% Mono-OH	11.5	0.38	9.16	0.39	0.79	0.002
% Di-OH	76.6	0.50	72.7	0.65	0.95	0.001
% Tri-OH	9.58	0.33	13.1	0.43	1.37	0.000
Total 12α-OH	3.62	0.17	8.35	0.83	2.30	0.001
Total non-12α-OH	5.67	0.20	46.2	4.68	8.15	0.000
12α-OH/non12α-OH	0.65	0.02	0.33	0.01	0.51	0.000
CA/CDCA	0.24	0.01	0.24	0.02	1.00	0.625
% 12α-OH	36.7	0.62	22.1	0.54	0.60	0.000
% non-12α-OH	63.3	0.62	77.9	0.54	1.23	0.000
Total Primary	3.34	0.15	27.2	2.59	8.15	0.000
Total Secondary	5.95	0.23	27.4	3.52	4.59	0.000
Primary/ Secondary	0.69	0.03	2.52	0.22	3.63	0.000
% Primary	36.7	0.70	49.4	1.06	1.35	0.000
% Secondary	63.3	0.70	50.6	1.06	0.80	0.000

¹Other bile acids: Nor-deoxycholic acid, 12-oxo-chenodeoxycholic acid, 3-dehydrocholic acid, 6-oxo-lithocholic acid, 7-oxo-lithocholic acid, 12-oxo-lithocholic acid, isolithocholic acid, and isodeoxycholic acid.

NA: Not available; BA: Bile acids; G: Glycine; T: Taurine; CDCA: Chenodeoxycholic acid; CA: Cholic acid; LCA: Lithocholic acid; UDCA: Ursodeoxycholic acid; DCA: Deoxycholic acid; HDCA: Hyodeoxycholic acid; MDCA: Murideoxycholic acid; MCA: Muricholic acid; HCA: Hyocholic acid.

Table 5 Representative bile acids concentrations and indices in medium- vs low- model for end-stage liver disease patients

BA (µmol/L) or BA indices	Low-MELD		Medium-MELD		Medium- vs low-MELD
	mean	SE	mean	SE	Ratio	P value
Total BA	59.2	7.94	116	24.8	1.96	1.000
Total LCA	3.40	0.35	6.01	1.72	1.77	0.175
Total UDCA	24.4	5.34	18.6	6.30	0.76	0.172
Total CDCA	18.3	2.31	71.4	16.3	3.90	0.000
Total DCA	5.30	0.96	6.08	1.47	1.15	1.000
Total HDCA	0.01	0.00	0.01	0.00	0.61	1.000
Total MDCA	0.05	0.01	0.06	0.01	1.28	1.000
Total CA	2.80	0.48	10.6	4.45	3.79	0.000
Total MCA	3.58	0.57	2.15	0.46	0.60	0.210
Total HCA	0.25	0.04	0.86	0.36	3.48	0.002
% LCA	9.31	0.53	7.97	1.47	0.86	1.000
% UDCA	23.1	1.29	14.3	2.52	0.62	1.000
% CDCA	34.7	1.21	55.6	3.17	1.60	0.000
% DCA	13.8	0.65	7.18	1.33	0.52	0.005
% HDCA	0.04	0.01	0.01	0.00	0.32	0.661
% MDCA	0.29	0.04	0.13	0.03	0.43	1.000
% CA	5.75	0.30	8.70	1.25	1.51	0.145
% MCA	7.15	0.48	3.70	0.91	0.52	0.000
% HCA	0.46	0.07	0.75	0.15	1.61	0.148
Total Unamidated	4.24	0.55	2.87	0.72	0.68	0.062
Total G-amidated	48.4	6.89	92.8	19.7	1.92	1.000
Total T-amidated	6.58	1.04	20.7	7.30	3.15	0.040
% Amidation	86.7	0.87	94.4	1.28	1.09	0.005
% G-amidation	75.5	0.96	77.2	2.73	1.02	1.000
% T-amidation	11.2	0.64	17.1	2.15	1.53	0.002
Total unsulfated	6.99	0.93	9.04	2.42	1.29	1.000
Total sulfated	52.3	7.21	107	23.2	2.05	1.000
% Sulfation	82.4	0.81	88.3	1.34	1.07	0.009
Total mono-OH	3.40	0.35	6.01	1.72	1.77	0.175
Total di-OH	48.1	7.01	96.2	20.9	2.00	1.000
Total tri-OH	6.63	0.90	13.6	4.90	2.06	0.301
% Mono-OH	9.31	0.53	7.97	1.47	0.86	1.000
% Di-OH	72.0	0.90	77.2	2.14	1.07	0.058
% Tri-OH	13.4	0.59	13.1	1.40	0.98	0.274
Total 12α-OH	8.55	1.23	16.8	4.86	1.96	0.053
Total non-12α-OH	50.7	7.21	99.6	21.5	1.96	1.000
12α-OH/non12α-OH	0.30	0.01	0.20	0.02	0.68	0.135
CA/CDCA	0.21	0.01	0.17	0.03	0.81	1.000
% 12α-OH	21.0	0.69	16.1	1.44	0.77	0.008
% non-12α-OH	79.0	0.69	83.9	1.44	1.06	0.008
Total primary	25.0	3.08	85.1	19.5	3.41	0.000
Total secondary	34.3	5.78	31.3	8.05	0.91	0.316
Primary/secondary	2.19	0.24	5.02	1.16	2.29	1.000
% Primary	48.1	1.40	68.7	3.10	1.43	0.014
% Secondary	51.9	1.40	31.3	3.10	0.60	0.014

BA: Bile acids; MELD: Model for end-stage liver disease; G: Glycine; T: Taurine; CDCA: Chenodeoxycholic acid; CA: Cholic acid; LCA: Lithocholic acid; UDCA: Ursodeoxycholic acid; DCA: Deoxycholic acid; HDCA: Hyodeoxycholic acid; MDCA: Murideoxycholic acid; MCA: Muricholic acid; HCA: Hyocholic acid.

Table 6 Representative bile acids concentrations and indices in compensated vs decompensated patients

BA (µmol/L) or BA indices	Compensated		Decompensated		Decompensated vs compensated
	mean	SE	mean	SE	Ratio	P value
Total BA	66.6	10.8	86.9	14.9	1.31	0.160
Total LCA	3.73	0.54	4.26	0.70	1.14	0.547
Total UDCA	27.0	6.49	21.0	9.82	0.78	0.687
Total CDCA	20.4	3.42	45.0	6.28	2.20	0.001
Total DCA	6.85	1.76	4.93	0.73	0.72	0.394
Total HDCA	0.01	0.00	0.02	0.01	1.61	0.430
Total MDCA	0.06	0.01	0.05	0.01	0.86	0.619
Total CA	2.62	0.40	6.28	1.51	2.39	0.024
Total MCA	4.48	0.93	4.07	0.83	0.91	0.864
Total HCA	0.20	0.04	0.64	0.14	3.23	0.002
% LCA	9.00	0.64	6.61	0.64	0.73	0.020
% UDCA	24.9	1.97	12.0	1.32	0.48	0.007
% CDCA	33.2	1.62	54.74	2.05	1.65	0.000
% DCA	14.3	0.98	9.17	1.00	0.64	0.000
% HDCA	0.02	0.01	0.03	0.01	1.42	0.532
% MDCA	0.34	0.14	0.11	0.01	0.33	0.264
% CA	6.07	0.54	7.58	0.48	1.25	0.262
% MCA	7.26	0.72	7.21	0.82	0.99	0.542
% HCA	0.35	0.05	0.74	0.08	2.09	0.005
Total unamidated	4.35	0.69	3.88	1.04	0.89	0.876
Total G-amidated	56.2	9.81	70.5	12.5	1.25	0.240
Total T-amidated	5.97	0.79	12.6	2.58	2.11	0.010
% Amidation	87.9	1.15	93.6	0.75	1.06	0.003
% G-amidation	76.5	1.30	78.8	1.23	1.03	0.161
% T-amidation	11.5	0.93	14.8	1.02	1.29	0.161
Total unsulfated	7.84	1.19	9.53	1.85	1.22	0.310
Total sulfated	58.7	9.97	77.4	13.4	1.32	0.156
% Sulfation	82.7	1.12	85.2	0.99	1.03	0.054
Total mono-OH	3.73	0.54	4.26	0.70	1.14	0.547
Total di-OH	54.4	9.55	70.9	13.1	1.31	0.174
Total tri-OH	7.30	1.25	11.0	1.96	1.51	0.085
% Mono-OH	9.00	0.64	6.61	0.64	0.73	0.020
% Di-OH	72.7	1.14	76.0	1.31	1.05	0.016
% Tri-OH	13.7	0.92	15.5	0.95	1.13	0.674
Total 12α-OH	10.1	2.08	11.44	1.75	1.14	0.554
Total non-12α-OH	56.5	9.36	75.51	14.0	1.34	0.137
12α-OH/non12α-OH	0.33	0.02	0.24	0.02	0.71	0.002
CA/CDCA	0.21	0.02	0.17	0.01	0.79	0.043
% 12α-OH	22.0	1.07	17.3	0.99	0.79	0.001
% non-12α-OH	78.0	1.07	82.7	0.99	1.06	0.001
Total primary	27.7	4.46	56.0	7.59	2.02	0.001
Total secondary	38.8	7.43	31.0	10.3	0.80	0.874
Primary/secondary	2.27	0.44	5.98	0.69	2.64	0.001
% Primary	46.9	2.05	70.3	1.88	1.50	0.000
% Secondary	53.1	2.05	29.7	1.88	0.56	0.000

BA: Bile acids; G: Glycine; T: Taurine; CDCA: Chenodeoxycholic acid; CA: Cholic acid; LCA: Lithocholic acid; UDCA: Ursodeoxycholic acid; DCA: Deoxycholic acid; HDCA: Hyodeoxycholic acid; MDCA: Murideoxycholic acid; MCA: Muricholic acid; HCA: Hyocholic acid.

Table 7 Univariate logistic regression analysis of bile acids concentrations and indices¹

BA (µmol/L) or BA indices	B value (regression coefficient)	P value	Exp(B)-odds ratio
			1-unit change	10% change	20% change
Total BA	0.080	0.000	1.08	1.08	1.16
Total CDCA	0.226	0.000	1.25	1.06	1.12
Total CA	1.181	0.000	3.26	1.05	1.11
% DCA	-0.080	0.000	0.92	0.78	0.61
% HDCA	-1.898	0.069	0.15	0.99	0.97
% MDCA	-0.174	0.162	0.84	0.99	0.98
Total G-amidated	0.084	0.000	1.09	1.07	1.14
Total unsulfated	0.784	0.000	2.19	1.08	1.16
Total sulfated	0.080	0.000	1.08	1.07	1.14
Total di-OH	0.094	0.000	1.10	1.07	1.15
Total tri-OH	0.731	0.000	2.08	1.06	1.13
Total non-12α-OH	0.146	0.000	1.16	1.09	1.18
12α-OH/non12α-OH	-2.349	0.000	0.10	0.86	0.74
% 12α-OH	-0.079	0.000	0.92	0.75	0.56
% non-12α-OH	0.079	0.000	1.08	1.65	2.72
Total primary	0.190	0.000	1.21	1.07	1.14
Primary/secondary	0.834	0.000	2.30	1.06	1.12
% Primary	0.033	0.000	1.03	1.13	1.27
% Secondary	-0.033	0.000	0.97	0.81	0.66

¹Bile acids with receiver operating characteristic (ROC)-areas under the ROC curve > 0.7 were included in this table.

BA: Bile acids; G: Glycine; CDCA: Chenodeoxycholic acid; CA: Cholic acid; DCA: Deoxycholic acid; HDCA: Hyodeoxycholic acid; MDCA: Murideoxycholic acid.

Table 8 Bile acids concentrations and indices in controls and patients with specific liver disease subtype¹

BA (µmol/L) or BA indices	Controls		Hepatitis C		Hepatitis B		Laennec cirrhosis		Primary biliary cholangitis		Primary sclerosing cholangitis		Autoimmune Hepatitis		α-1 antitrypsin deficiency		NASH		Carcinoma		Cryptogenic cirrhosis		Elevated LFT
	n = 103		n = 71		n = 15		n = 117		n = 12		n = 17		n = 27		n = 6		n = 56		n = 26		n = 11		n = 22
	mean	SE	mean	SE	mean	SE	mean	SE	mean	SE	mean	SE	mean	SE	mean	SE	mean	SE	mean	SE	mean	SE	mean	SE
Total BA	9.30	0.33	53.3a	9.96	13.7a	5.23	62.0a	9.44	237a	69.8	124a	27.4	71.9a	15.2	25.4a	6.79	29.8a	4.31	90.9a	26.7	56.0a	16.3	106a	69.3
Total CDCA	2.52	0.13	27.0a	4.89	6.76a	4.16	30.0a	4.25	28.6a	9.99	39.4a	10.7	29.2a	8.46	9.14a	3.12	13.6a	2.33	31.9a	8.76	27.7a	7.39	31.3a	18.8
Total CA	0.44	0.03	3.05a	0.54	1.16a	0.70	4.00a	1.02	5.07a	2.27	6.47a	2.25	1.96a	0.39	2.44a	0.85	1.65a	0.23	3.54a	1.11	2.70a	0.59	5.55a	3.09
% DCA	31.1	0.68	16.2a	1.27	19.9a	3.26	13.1a	0.95	7.99a	2.22	9.01a	1.86	15.6a	1.43	18.9	4.23	15.7a	1.22	14.9a	1.43	7.93a	2.68	17.4a	3.07
% HDCA	0.07	0.01	0.02a	0.00	0.06	0.02	0.02a	0.01	0.03	0.02	0.01	0.00	0.02	0.00	0.03	1.21	0.05	0.01	0.06	0.02	0.04	0.03	0.05	0.04
% MDCA	0.64	0.04	0.19a	0.04	0.38	0.08	0.16a	0.03	0.15a	0.07	0.18a	0.06	0.22a	0.05	0.38	0.16	0.49	0.21	0.16a	0.05	0.07a	0.01	1.34	0.98
Total G-amidated	7.59	0.29	44.8a	9.11	11.6a	4.52	50.8a	8.26	210a	60.4	106a	24.0	61.8a	13.5	16.3a	4.58	26.0a	4.01	78.1a	24.5	49.2a	14.9	86.8a	58.2
Total unsulfated	0.94	0.05	7.69a	1.43	1.62a	0.38	7.94a	1.21	17.6a	8.66	6.21a	1.13	6.06a	1.44	3.82a	1.21	4.64a	0.65	13.0a	3.16	3.58a	0.58	13.0a	9.22
Total sulfated	8.35	0.31	45.6a	8.73	12.1a	4.96	54.1a	8.59	219a	62.0	117a	26.5	65.9a	14.7	21.6a	6.11	25.2a	4.04	77.9a	24.2	52.4a	16.0	92.9a	60.2
Total di-OH	7.30	0.29	41.0a	7.98	10.6a	4.41	49.05a	7.97	214a	62.50	111a	25.9	60.9a	13.7	15.9a	4.72	23.2a	3.87	71.9a	23.0	50.3a	16.0	91.0a	61.0
Total tri-OH	0.82	0.04	8.61a	1.63	1.82a	0.80	8.59a	1.54	12.48a	6.02	9.28a	2.44	4.83a	0.76	5.24a	1.83	4.27a	0.66	13.8a	3.62	3.92a	0.65	10.8a	6.50
Total non-12α-OH	5.67	0.20	41.8a	7.46	10.3a	4.52	50.8a	7.85	224a	66.9	113a	26.5	62.3a	14.2	19.4a	5.59	23.8a	3.74	75.7a	22.1	51.1a	16.1	94.7a	66.1
12α-OH/non12α-OH	0.65	0.02	0.37a	0.05	0.51	0.08	0.31a	0.02	0.15a	0.05	0.29a	0.05	0.33a	0.04	0.40	0.07	0.34a	0.02	0.28a	0.03	0.22a	0.05	0.37a	0.06
% 12α-OH	36.7	0.62	22.8a	1.28	31.1	2.9	21.3a	1.0	10.8a	2.69%	18.8a	2.6	22a	1.69	27.1	3.18	23.5a	1.14	20.6a	1.55	16.2a	2.88	24.8a	2.8
% non-12α-OH	63.3	0.62	77.2a	1.28	68.9	2.9	78.7a	1.0	89.2a	2.69	81.2a	2.6	78.0a	1.69	72.9	3.18	76.5a	1.14	79.4a	155	83.8a	2.88	75.2a	2.8
Total primary	3.34	0.15	35.6a	6.23	8.58a	4.93	38.6a	5.47	41.1a	15.6	48.6a	12.6	34.1a	8.91	14.4a	4.33	17.9a	2.70	45.7a	12.1	31.6a	7.89	42.1a	23.9
Primary/secondary	0.69	0.03	3.70a	0.55	1.52a	0.59	4.33a	0.60	0.30	0.10	2.88a	0.68	2.09a	0.60	1.26	0.26	2.28a	0.30	2.32a	0.40	6.43a	2.09	1.70a	0.43
% Primary	36.7	0.70	60.2a	1.99	47.0a	3.93	60.9a	1.7	18.0a	2.97	51.6a	5.09	50.1a	2.77	50.3	5.43	52.8a	2.21	56.1a	3.25	68.2a	5.80	49.3a	4.6
% Secondary	63.3	0.70	39.8a	1.99	53.0a	3.93	39.1a	1.7	82.0a	2.97	48.4a	5.09	49.9 a	2.77	49.7	5.43	47.2a	2.21	43.9a	3.25	31.8a	5.80	50.7a	4.6

¹Bile acids with receiver operating characteristic (ROC)-areas under the ROC curve > 0.7 were included in this table.

^aSignificant difference between each specific liver disease subtype vs controls (P < 0.05).

BA: Bile acids; G: Glycine; CDCA: Chenodeoxycholic acid; CA: Cholic acid; DCA: Deoxycholic acid; HDCA: Hyodeoxycholic acid; MDCA: Murideoxycholic acid.

Table 9 Summary of non-bile acids parameters

Non-BA parameters	Controls		Patients										ROC1
			Pooled		Low-MELD		Medium-MELD		Compensated		Decompensated		AUC
	mean	SE	mean	SE	mean	SE	mean	SE	mean	SE	mean	SE
Creatinine (mg/dL)	0.87	0.01	0.99	0.05	0.93	0.07	1.33b	0.16	1.05	0.15	1.05	0.06	0.539
Albumin (g/dL)	3.96	0.02	3.61a	0.03	3.61	0.03	2.82b	0.10	3.69	0.04	3.03c	0.06	0.713
INR	0.99	0.01	1.18a	0.02	1.11	0.01	1.63b	0.10	1.15	0.03	1.36c	0.03	0.758
Protime (s)	13.4	0.10	10.2a	0.33	13.6	0.13	19.4b	0.98	11.2	0.52	13.7c	0.64	0.591
AST (U/L)	22.8	0.34	53.2a	2.31	52.1	2.59	79.2b	10.4	52.6	3.97	61.7	4.85	0.876
ALT (U/L)	21.0	0.46	51.0a	2.60	51.0	3.24	46.0	5.54	49.0	4.09	40.6	3.55	0.825
Bilirubin (mg/dL)	0.62	0.03	1.58a	0.09	1.31	0.05	5.02b	0.68	1.42	0.12	3.04c	0.29	0.804
AST/ALT	1.15	0.01	1.22	0.02	1.21	0.03	1.79b	0.09	1.21	0.04	1.61c	0.05	0.500
MELD	7.13	0.10	10.3a	0.24	9.07	0.16	18.9	0.42	9.54	0.37	14.0c	0.46	0.747
APRI	NA	NA	0.93	0.06	1.05	0.07	2.44b	0.42	0.94	0.08	1.63c	0.18	NA

¹Areas under the receiver operating characteristic curve from receiver operating characteristic analysis of pooled patients vs controls.

^aSignificant difference between patients vs controls (P < 0.05).

^bSignificant difference between medium-model for end-stage liver disease vs low-model for end-stage liver disease groups (P < 0.05).

^cSignificant difference between decompensated vs compensated patients (P < 0.05).

NA: Not available; BA: Bile acids; MELD: Model for end-stage liver disease; INR: International normalized ratio; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; APRI: Aspartate aminotransferase/platelet ratio index; ROC: Receiver operating characteristic curve; AUC: Areas under the receiver operating characteristic curve.