Plasma Metabolic Profiling Analysis of Gout Party on Acute Gout Arthritis Rats Based on UHPLC–Q–TOF/MS Combined with Multivariate Statistical Analysis
Abstract
:1. Introduction
2. Results
2.1. Histopathological Evaluation
2.2. Determination of IL-1β and IL-6 in Plasma.
2.3. Chemical Characterization of Gout Party
2.4. Multivariate Data Analysis
2.5. Optimization via ROC Curve Analysis
3. Discussion
3.1. Amino Acid Metabolism
3.2. Fatty Acid Metabolism
3.3. Bile Acid Metabolism
3.4. Energy Metabolism
4. Materials and Methods
4.1. Reagents and Materials
4.2. Extraction of Gout Party
4.3. Animals and Experimental Method
4.4. Sample Collection and Preparation
4.5. Metabolomic Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
GA | Gouty arthritis |
AGA | Acute gouty arthritis |
UPLC–Q–TOF/MS | Ultra-high-performance liquid chromatography–tandem quadrupole/time-of-flight mass spectrometry |
PCA | Principal component analysis |
PLS-DA | partial least square discrimination analysis |
SUCs | Sodium urate crystals |
SUA | Serum uric acid |
TCM | Traditional Chinese medicine |
ELISA | Enzyme–linked immunosorbent assay |
QC | Quality control |
BPI | base peak intensity |
ROC | receiver operating characteristic |
PCS | p–cresol sulfate |
GPH | Gout Party high–dose group |
GPL | Gout Party low–dose group |
VIP | Variable importance in projection |
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Experiment Name | RSD (Peak Area) | RSD (Retention Time) |
---|---|---|
Instrument precision | <14.3% | <1.0% |
Method repeatability | <13.7% | <1.0% |
Sample stability | <14.8% | <1.0% |
No. | tR (min) | m/z Obsd | m/z Calcd | Fromula | Identification | Parention | MS/MS |
---|---|---|---|---|---|---|---|
1 | 5.19 | 461.1674 | 461.1659 | C20H30O12 | Decaffeoyl-verbascoside | [M–H]− | 161.04;135.04 |
2 | 5.86 | 486.2701 | 486.2703 | C24H39NO9 | Mesaconine | [M+H]+ | 468.25;454.24 |
3 | 6.17 | 387.1321 | 387.1291 | C17H24O10 | Genipin | [M–H]− | 225.07 |
4 | 6.64 | 454.2778 | 454.2805 | C24H39NO7 | Fuziline | [M+H]+ | 436.28;404.28 |
5 | 6.86 | 358.2365 | 358.2382 | C22H31NO3 | Songorine | [M+H]+ | 340.23 |
6 | 7.33 | 500.2831 | 500.286 | C25H41NO9 | Aconine | [M+H]+ | 482.27;468.25 |
7 | 8.03 | 523.1666 | 523.1663 | C21H32O15 | Miltide | [M–H]− | 463.14;343.10 |
8 | 8.58 | 470.2725 | 470.2754 | C24H39NO8 | Hypaconine | [M+H]+ | 438.24;406.22 |
9 | 8.76 | 454.2769 | 454.2805 | C24H39NO7 | Delectinine | [M+H]+ | 436.26;404.24 |
10 | 9.26 | 438.2823 | 438.2856 | C24H39NO6 | Neoline | [M+H]+ | 420.27;388.24 |
11 | 9.64 | 468.2935 | 468.2961 | C25H41NO7 | Browniine | [M+H]+ | 450.28;436.26 |
12 | 9.94 | 468.2935 | 468.2961 | C25H41NO7 | Lycoctonine | [M+H]+ | 362.29; 393.30 |
13 | 10.59 | 422.2890 | 422.2906 | C24H39NO5 | Talatizamine | [M+H]+ | 390.26;372.25 |
14 | 12.07 | 452.3019 | 452.3012 | C25H41NO6 | Chasmanine | [M+H]+ | 420.27;388.24 |
15 | 14.26 | 606.2914 | 606.2914 | C31H43NO11 | 10-OH-Benzoylmesaco-nine | [M+H]+ | 588.27;574.26;556.25 |
16 | 15.99 | 623.1956 | 623.1976 | C29H36O15 | Acteoside | [M–H]− | 461.16 161.02 |
17 | 17.44 | 576.2809 | 576.2809 | C30H41NO10 | Unknown | [M+H]+ | 558.27;544.25;526.24 |
18 | 19.01 | 544.2915 | 544.2910 | C30H41NO8 | Gadenine | [M+H]+ | 512.26;494.25;484.23 |
19 | 21.12 | 604.3154 | 604.3122 | C32H45NO10 | Benzoylaconitine | [M+H]+ | 588.31 |
20 | 22.26 | 572.2848 | 572.2860 | C31H41NO9 | Pyromesaconitine | [M+H]+ | 554.27;540.25;522.24 |
21 | 22.35 | 574.3044 | 574.3016 | C31H43NO9 | BenzoylhypaconineS | [M+H]+ | 542.27;510.24 |
22 | 22.59 | 516.2947 | 516.2961 | C29H41NO7 | Unknown | [M+H]+ | 442.25;414.26 |
23 | 23.31 | 616.3135 | 616.3122 | C33H45NO10 | Pyrojesaconitine | [M+H]+ | 598.30;584.28;566.27 |
24 | 23.78 | 558.3096 | 558.3067 | C31H43NO8 | Deoxybenzoylhypaconitine | [M+H]+ | 526.27;508.27 |
25 | 23.88 | 648.3031 | 648.3020 | C33H45NO12 | Beiwutine | [M+H]+ | 588.29 |
26 | 24.95 | 556.2905 | 556.2910 | C31H41NO8 | Pyrohypaconitine | [M+H]+ | 524.26;492.23 |
27 | 25.25 | 586.3019 | 586.3016 | C32H43NO9 | Pyroaconitine | [M+H]+ | 568.29;554.27;472.21 |
28 | 25.64 | 632.3107 | 632.3071 | C33H45NO11 | 16-O-demethylaconitine | [M+H]+ | 572.28;540.25;508.22 |
29 | 25.64 | 632.3107 | 632.3071 | C33H45NO11 | Mesaconitine | [M+H]+ | 572.30;540.30 |
30 | 27.00 | 570.3082 | 570.3067 | C32H43NO8 | Pyrodeoxyaconitine | [M+H]+ | 538.27;506.25;478.22 |
31 | 27.11 | 616.3145 | 616.3122 | C33H45NO10 | HypaconitineS | [M+H]+ | 584.28;556.28;524.26 |
32 | 27.66 | 628.3131 | 628.3122 | C34H45NO10 | Anhydroaconitine | [M+H]+ | 568.28;536.26;508.26 |
33 | 28.33 | 700.2738 | 700.2758 | C39H41NO11 | Trifoliolasine E | [M+H]+ | 640.25;578.23 |
34 | 28.55 | 630.3281 | 630.3278 | C34H47NO10 | Deoxyaconitine | [M+H]+ | 598.30;570.30;538.27 |
35 | 28.74 | 505.3532 | 505.3529 | C30H48O6 | 16-Oxo-alisol A | [M+H]+ | 505.33;487.34 |
36 | 29.32 | 529.3539 | 529.3529 | C32H48O6 | Alisol C-23-acetate | [M+H]+ | 551.33;511.34 |
37 | 30.92 | 455.3502 | 455.3525 | C30H46O3 | Alisol I | [M+H]+ | 477.33;543.33 |
38 | 31.67 | 487.3411 | 487.3423 | C30H46O5 | Alisol C | [M+H]+ | 509.33;469.32 |
39 | 31.71 | 489.3563 | 489.3580 | C30H48O5 | Alisol F | [M+H]+ | 511.33;471.34 |
40 | 32.51 | 471.3463 | 471.3474 | C30H46O4 | Alisol H | [M+H]+ | 493.32;453.33 |
41 | 33.98 | 515.3711 | 515.3736 | C32H50O5 | Alisol B-23-acetate | [M+H]+ | 537.35;497.35 |
No. | tR(min) | m/z Obsd | m/z Calcd | MS/MS | PPM | Metabolites | Formula | Content Variance | |||
---|---|---|---|---|---|---|---|---|---|---|---|
Model | High | Low | Control | ||||||||
1 | 0.79 | 101.0234 | 101.0239 | 53.00, 83.01 | −4.95 | 2–Ketobutyric acid | C4H6O3 | ↓** | ↑ | ↑ | ↑ |
2 | 0.79 | 143.0344 | 143.0344 | 59.01 81.03 | 0.00 | 3–Hexenedioic acid | C6H8O4 | ↓** | ↑ | ↑ | ↑ |
3 | 0.79 | 85.0288 | 85.0290 | 67.01 68.99 | –2.35 | But–2–enoic acid | C4H6O2 | ↓** | ↓ | ↑ | ↑ |
4 | 0.81 | 174.0878 | 174.0879 | 100.08 112.09 | −0.57 | Argininic acid | C6H13N3O3 | ↓** | ↑ | ↓ | ↑ |
5 | 0.81 | 195.0487 | 195.0505 | 89.02 | −9.23 | Galactonic acid | C6H12O7 | ↓** | ↑ | ↑ | ↑ |
6 | 0.84 | 88.0394 | 88.0399 | 88.04 | −5.68 | Alanine | C3H7NO2 | ↑** | ↑ | ↓ | ↑ |
7 | 0.91 | 119.0342 | 119.0344 | 87.01 89.02 101.02 | −1.68 | Erythrulose | C4H8O4 | ↑* | ↓ | ↓ | ↓ |
8 | 0.92 | 89.0235 | 89.0239 | 71.01 73.00 89.02 | −4.49 | Lactic acid | C3H6O3 | ↓** | ↑ | ↓ | ↑ |
9 | 2.36 | 158.0814 | 158.0817 | 59.01 84.99 101.02 | –1.90 | 3–Dehydrocarnitine | C7H13NO3 | ↑** | ↓ | ↑ | ↓ |
10 | 2.41 | 192.0658 | 192.0661 | 65.04 74.02 77.04 | −1.56 | m–Methylhippuric acid | C10H11NO3 | ↑* | ↑ | ↑# | ↑ |
11 | 2.52 | 417.1171 | 417.1186 | 103.04 121.03 | −3.60 | Equol 4’–O–glucuronide | C21H22O9 | ↓** | ↓ | ↑## | ↓ |
12 | 2.88 | 159.1016 | 159.1021 | 59.01 71.03 87.01 | −3.14 | 3–Hydroxyoctanoic acid | C8H16O3 | ↑* | ↑ | ↑ | ↑ |
13 | 2.95 | 187.0059 | 187.0065 | 51.02 75.02 77.04 | −3.21 | p–Cresol sulfate | C7H8O4S | ↑* | ↓ | ↑# | ↑ |
14 | 3.08 | 448.3050 | 448.3063 | 74.02 116.04 | −2.90 | Deoxycholic acid glycine conjugate | C26H43NO5 | ↓* | ↓ | ↓## | ↓## |
15 | 3.09 | 465.3028 | 465.3039 | 116.04 373.27 | −2.36 | Glycocholic acid | C27H46O4S | ↓** | ↓ | ↑# | ↑# |
16 | 4.78 | 380.2552 | 380.2566 | 78.96 96.97 | −3.68 | Sphinganine 1–phosphate | C18H40NO5P | ↓** | ↓ | ↓ | ↓## |
17 | 5.69 | 502.2945 | 502.2934 | 78.96 140.01 153.00 | 2.19 | LPE (0:0/20:3) | C25H46NO7P | ↓** | ↑ | ↑ | ↑ |
18 | 5.76 | 452.2775 | 452.2777 | 98.99 140.01 196.04 | –0.44 | LPE (0:0/16:0) | C21H44NO7P | ↓* | ↑ | ↓ | ↑ |
19 | 7.04 | 480.3089 | 480.3090 | 78.96 196.04 265.25 | −0.21 | LPC (15:0) | C23H48NO7P | ↓** | ↑ | ↓# | ↑## |
20 | 7.75 | 543.1513 | 543.1536 | 59.01 87.01 89.02 | −4.24 | Estriol 3–sulfate 16–glucuronide | C24H32O12S | ↑* | ↓ | ↑ | ↑ |
21 | 7.76 | 395.2190 | 395.2199 | 62.96 96.97 113.13 | −2.28 | 10–Hydroxy–9–(phosphonooxy)octadecanoate | C18H37O7P | ↑** | ↓ | ↑ | ↑ |
22 | 7.76 | 327.2320 | 327.2324 | 127.08 161.13 163.15 | –1.22 | Docosahexaenoic acid | C22H32O2 | ↑** | ↓ | ↓ | ↑ |
Groups | Administration | Dosage | Mode of Administration | Time | N |
---|---|---|---|---|---|
Blank | Saline | 15 mL/kg | Gavage | 7 days | 6 |
Model | Saline | 15 mL/kg | Gavage | 7 days | 6 |
Control | Colchicine | 0.8 mg/kg | Gavage | 7 days | 6 |
GPH | Gout Party | 20 g/kg | Gavage | 7 days | 6 |
GPL | Gout Party | 1 0g/kg | Gavage | 7 days | 6 |
T (min) | Phase A (0.1% Formic Acid in Water) | Phase B (0.1% Formic Acid Inacetonitrile) |
---|---|---|
0 | 99.0 | 1.0 |
0.5 | 99.0 | 1.0 |
2 | 50.0 | 50.0 |
9 | 1.0 | 99.0 |
10 | 1.0 | 99.0 |
11 | 99.0 | 1.0 |
13 | 99.0 | 1.0 |
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Wang, Y.; Bi, C.; Pang, W.; Liu, Y.; Yuan, Y.; Zhao, H.; Zhang, T.; Zhao, Y.; Li, Y. Plasma Metabolic Profiling Analysis of Gout Party on Acute Gout Arthritis Rats Based on UHPLC–Q–TOF/MS Combined with Multivariate Statistical Analysis. Int. J. Mol. Sci. 2019, 20, 5753. https://doi.org/10.3390/ijms20225753
Wang Y, Bi C, Pang W, Liu Y, Yuan Y, Zhao H, Zhang T, Zhao Y, Li Y. Plasma Metabolic Profiling Analysis of Gout Party on Acute Gout Arthritis Rats Based on UHPLC–Q–TOF/MS Combined with Multivariate Statistical Analysis. International Journal of Molecular Sciences. 2019; 20(22):5753. https://doi.org/10.3390/ijms20225753
Chicago/Turabian StyleWang, Yuming, Chenghao Bi, Wentao Pang, Yuechen Liu, Yu Yuan, Huan Zhao, Tianpu Zhang, Yungang Zhao, and Yubo Li. 2019. "Plasma Metabolic Profiling Analysis of Gout Party on Acute Gout Arthritis Rats Based on UHPLC–Q–TOF/MS Combined with Multivariate Statistical Analysis" International Journal of Molecular Sciences 20, no. 22: 5753. https://doi.org/10.3390/ijms20225753