A rapid and simple method was established to compare the changes of chemical constituents in the processing of Polygonum multiflorum thumb (PM). Primary and secondary metabolites of PM were detected and identified by1 H-NMR spectroscopy. Multivariate analysis method (OPLS-DA) is used for grouping and searching for potential biomarkers. Twenty-six metabolites (including fourteen potential biomarkers) were identified in samples from different processing methods. The results suggest that the content changes of fourteen potential biomarkers (including Valine, Ethyl alcohol, Threonine, Lactic acid, Alanine, Acetic acid, Proline, Succinic acid, Lysine, Choline, Sucrose, Glycine, 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside, β-D-glucoside, Gallic acid, 5-HF) in different processing methods of PM can influence on the pharmacological activity in vivo and to provide reference for proper use of PM .
Table 1H NMR chemical shifts of metabolites identified in the PM samples.
No. |
Metabolites |
Chemical shift |
Correlated NMR spectra |
p-value |
1 |
Leucine |
0.97(t),3.742(m) |
HSQC,J-res |
|
2 |
Isoleucine |
0.95(t),1.02(d) |
COSY,HSQC,J-res |
|
3 |
Valine* |
1.00(d),1.05(d),2.29(m),3.63(d) |
COSY,HSQC,J-res,HMBC |
<0.05 |
4 |
Ethanol ** |
1.22(t), 3.68 |
J-res, COSY, HSQC, HMBC |
<0.001 |
5 |
Threonine* |
1.36(d),3.62(d),4.38(m) |
COSY,HSQC,J-res, HMBC |
<0.05 |
6 |
Lactate* |
1.37(d),4.10(q) |
COSY,HSQC,J-res |
<0.001 |
7 |
Alanine** |
1.52(d), 3.82 |
J-res, COSY |
<0.001 |
8 |
arginine |
1.94(m), 3.22, 3.59 |
COSY, J-res |
|
9 |
Acetate** |
1.96(s) |
HSQC,J-res, HMBC |
<0.001 |
10 |
Proline* |
2.00(m), 2.10(m), 2.36(m),4.16(dd) |
COSY,HSQC,J-res. HMBC |
<0.05 |
11 |
Succinate** |
2.45(s) |
J-res, HSQC, HMBC |
<0.001 |
12 |
Malate |
2.36(dd), 2.71(dd), 4.36(dd) |
COSY,HSQC, HMBC,TOCSY |
|
13 |
γ-aminobutyrate * |
3.05(t), 1.92, 2.32 |
COSY,HSQC,J-res,HMBC |
<0.001 |
14 |
α-ketoglutaric acid |
3.05(t), 2.40 |
COSY, HSQC,J-res |
|
15 |
Malonate |
3.16(s) |
HSQC, HMBC, J-res |
|
16 |
Choline* |
3.18(s) |
J-res, HSQC, HMBC |
<0.05 |
17 |
Sucrose** |
3.51(t),3.60(dd),3.72(s),4.09(t),4.25(d),5.45(d) |
COSY,HSQC,TOCSY,HMBC |
<0.001 |
18 |
Glycine* |
3.60(S) |
J-res |
<0.05 |
19 |
Fructose |
4.07(dd) |
COSY,HSQC,J-res |
|
20 |
TSG** |
4.40(d),6.34(d),6.56(d),6.71(d),6.78(d),7.23(d),7.30(d),3.8(m) |
COSY,HSQC,HMBC,TOCSY |
<0.001 |
21 |
β -glucoside** |
4.69(d),3.29(dd) |
COSY,HSQC,HMBC,J-res |
<0.001 |
22 |
α -glucoside |
5.28(d),3.84(m) |
COSY,HSQC,HMBC,J-res |
|
23 |
Gallic acid** |
7.08(s) |
HSQC,J-res |
<0.001 |
24 |
Formic acid |
8.50(s) |
HSQC.J-res |
|
p- values are the result of T test
*(p<0.05) ** (p<0.001).
