Biomarker Profiling with Targeted Metabolomic Analysis of Plasma and Urine Samples in Patients with Type 2 Diabetes Mellitus and Early Diabetic Kidney Disease
Abstract
:1. Introduction
2. Materials and Methods
2.1. Patients and Compliance with Ethical Standards
2.2. Chemicals and Reagents
2.3. Sample Preparation
2.4. UHPLC-QTOF-ESI+-MS Analysis
2.5. Data Processing and Statistical Analysis
2.6. Metabolite Identification
2.7. Quantitative Evaluation
3. Results
3.1. Untargeted Multivariate and Univariate Analyses
3.2. Multivariate Analysis for Targeted Metabolites: Fold Change, p Values, and AUC
3.2.1. Calibrations and Validation Parameters
3.2.2. Quantitative Evaluation
4. Discussion
4.1. TRP Metabolic Pathway and Its Role in DKD
4.2. Assessment of Taurine as a Potential Biomarker in DKD
4.3. Association between Acetylcarnitine and the Presence of DKD
4.4. Glycine and Its Metabolite Tiglylglycine—Involvement in DKD
4.5. From Metabolic Studies to Clinical Practice
4.6. Other Potential Metabolites in Patients with DM
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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m/z | Identification | RT (min) | Plasma AUC | Urine AUC |
---|---|---|---|---|
76.0815 | Glycine | 0.9 | 0.868 | 0.926 |
114.0983 | Creatinine | 0.7 | 0.512 | 0.637 |
125.1042 | Taurine | 1.6 | 0.602 | 0.525 |
190.0625 | Kynurenic acid | 11.4 | 0.787 | 0.591 |
204.1369 | L-Acetylcarnitine | 1. | 0.556 | 0.550 |
205.1068 | L-Tryptophan | 2.8 | 0.618 | 0.788 |
158.106 | Tiglylglycine | 10.7 | - | 0.684 |
A | |||||
---|---|---|---|---|---|
m/z | Identification | f Value | p Value | FDR | Tukey’s HSD |
76.0815 | Glycine | 0.624 | 0.601 | 0.645 | P1-C; P2-C; P3-C; P2-P1; P3-P1; P3-P2 |
114.0983 | Creatinine | 2.188 | 0.094 | 0.281 | P1-C; P2-C; P3-C; P2-P1; P3-P1; P3-P2 |
125.1042 | Taurine | 0.984 | 0.403 | 0.605 | P1-C; P2-C; P3-C; P2-P1; P3-P1; P3-P2 |
190.0625 | Kynurenic acid | 4.395 | 0.006 | 0.035 | P1-C; P2-C; P3-C; P2-P1; P3-P1; P3-P2 |
204.1369 | L-Acetylcarnitine | 1.303 | 0.277 | 0.555 | P1-C; P2-C; P3-C; P2-P1; P3-P1; P3-P2 |
205.1068 | L-Tryptophan | 0.556 | 0.645 | 0.645 | P1-C; P2-C; P3-C; P2-P1; P3-P1; P3-P2 |
B | |||||
m/z | Identification | f Value | p Value | FDR | Tukey’s HSD |
76.0815 | Glycine | 104.260 | 0.0005 | 0.0003 | C-P1; C-P2; C-P3; P2-P1; P1-P3; P2-P3 |
114.0983 | Creatinine | 0.391 | 0.759 | 0.759 | C-P1; P2-C; P3-C; P2-P1; P3-P1; P2-P3 |
125.1042 | Taurine | 2.316 | 0.080 | 0.140 | C-P1; C-P2; C-P3; P1-P2; P3-P1; P3-P2 |
158.1060 | Tiglylglycine | 2.910 | 0.038 | 0.089 | C-P1; C-P2; C-P3; P2-P1; P3-P1; P3-P2 |
190.0625 | Kynurenic acid | 1.916 | 0.131 | 0.184 | C-P1; C-P2; P3-C; P1-P2; P3-P1; P3-P2 |
204.1369 | L-Acetylcarnitine | 1.032 | 0.382 | 0.445 | C-P1; C-P2; C-P3; P2-P1; P1-P3; P2-P3 |
205.1068 | L-Tryptophan | 5.988 | 0.001 | 0.003 | P1-C; P2-C; P3-C; P1-P2; P3-P1; P3-P2 |
Name | Linear Range μM | Curve Equation | R2 | LOD (μM) | LOQ (μM) |
---|---|---|---|---|---|
Glycine | 10–200 | y = 596.72x − 963.05 | 0.993 | 0.2 | 0.8 |
Creatinine—serum | 5–25 | y = 2693.3x − 254.5 | 0.994 | 0.5 | 1.0 |
Creatinine—urine | 200–2000 | y = 2765.8x − 73120 | 0.993 | 0.5 | 1.0 |
Taurine | 5–25 | y = 1479.6x + 543.2 | 0.996 | 0.5 | 1.0 |
Kynurenic acid | 0.1–2 | y = 24309x + 355.37 | 0.999 | 0.08 | 0.1 |
L-Acetylcarnitine | 1–5 | y = 36813x − 3879.2 | 0.994 | 0.2 | 0.8 |
L-Tryptophan | 3–50 | y = 820.31x − 981.3 | 0.998 | 0.8 | 1.00 |
Metabolite | Initial Concentration (µM) | Measured Concentration (µM) | Recovery (%) |
---|---|---|---|
Glycine | 20 | 17.5 | 87.5 |
Creatinine | 20 | 18.2 | 91.0 |
Taurine | 10 | 8.55 | 85.5 |
Kynurenic acid | 2 | 1.65 | 82.5 |
L-Acetylcarnitine | 2 | 1.85 | 92.5 |
L-Tryptophan | 10 | 9.15 | 91.5 |
IS (DOXO) | 1.4 | 1.25 | 89.3 |
Molecule | Control (n = 20) | P1 (n = 30) | P2 (n = 30) | P3 (n = 30) | ||||
---|---|---|---|---|---|---|---|---|
Mean | ±SD | Mean | ±SD | Mean | ±SD | Mean | ±SD | |
Glycine | 221.32 | 39.84 | 196.24 | 39.25 | 188.56 | 43.37 | 214.62 | 47.22 |
L-tryptophan | 56.59 | 10.19 | 51.38 | 10.28 | 49.53 | 11.39 | 54.51 | 11.99 |
Kynurenic acid | 5.81 | 1.05 | 4.80 | 0.96 | 4.50 | 1.04 | 4.32 | 0.95 |
Taurine | 86.04 | 15.49 | 85.98 | 17.20 | 75.99 | 17.48 | 80.34 | 17.67 |
L-Acetylcarnitine | 5.52 | 0.99 | 5.73 | 1.15 | 5.41 | 1.24 | 5.58 | 1.23 |
Creatinine (S) | 75.26 | 13.55 | 74.20 | 14.84 | 65.67 | 15.10 | 105.06 | 23.11 |
Molecule | Control (n = 20) | P1 (n = 30) | P2 (n = 30) | P3 (n = 30) | ||||
---|---|---|---|---|---|---|---|---|
Mean | ±SD | Mean | ±SD | Mean | ±SD | Mean | ±SD | |
Glycine | 114.89 | 20.68 | 87.36 | 17.47 | 87.43 | 20.11 | 111.39 | 24.51 |
Tiglylglycine (μM glycine units) | 22.08 | 3.97 | 17.45 | 3.49 | 16.69 | 3.84 | 29.77 | 6.55 |
L-tryptophan | 7.63 | 1.37 | 12.14 | 2.43 | 9.03 | 2.08 | 16.64 | 3.66 |
Kynurenic acid | 0.34 | 0.06 | 0.42 | 0.08 | 0.35 | 0.08 | 0.59 | 0.13 |
Taurine | 7.6 | 1.37 | 7.58 | 1.52 | 6.65 | 1.53 | 10.72 | 2.36 |
L-Acetylcarnitine | 0.78 | 0.14 | 0.65 | 0.13 | 0.63 | 0.14 | 0.56 | 0.12 |
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Mogos, M.; Socaciu, C.; Socaciu, A.I.; Vlad, A.; Gadalean, F.; Bob, F.; Milas, O.; Cretu, O.M.; Suteanu-Simulescu, A.; Glavan, M.; et al. Biomarker Profiling with Targeted Metabolomic Analysis of Plasma and Urine Samples in Patients with Type 2 Diabetes Mellitus and Early Diabetic Kidney Disease. J. Clin. Med. 2024, 13, 4703. https://doi.org/10.3390/jcm13164703
Mogos M, Socaciu C, Socaciu AI, Vlad A, Gadalean F, Bob F, Milas O, Cretu OM, Suteanu-Simulescu A, Glavan M, et al. Biomarker Profiling with Targeted Metabolomic Analysis of Plasma and Urine Samples in Patients with Type 2 Diabetes Mellitus and Early Diabetic Kidney Disease. Journal of Clinical Medicine. 2024; 13(16):4703. https://doi.org/10.3390/jcm13164703
Chicago/Turabian StyleMogos, Maria, Carmen Socaciu, Andreea Iulia Socaciu, Adrian Vlad, Florica Gadalean, Flaviu Bob, Oana Milas, Octavian Marius Cretu, Anca Suteanu-Simulescu, Mihaela Glavan, and et al. 2024. "Biomarker Profiling with Targeted Metabolomic Analysis of Plasma and Urine Samples in Patients with Type 2 Diabetes Mellitus and Early Diabetic Kidney Disease" Journal of Clinical Medicine 13, no. 16: 4703. https://doi.org/10.3390/jcm13164703