Metabolomic evaluation of Euphorbia pekinensis induced nephrotoxicity in rats

Figures & data

Figure 1. Clinical chemistry: the value of creatinine (CREA) and blood urea nitrogen (BUN) (n = 8). #Compared with HCG (p < 0.01).

Figure 2. Histopathological photomicrographs of rat kidney sections (40×).

Figure 3. PCA score plot (A) and corresponding loading plot (B) of kidney samples in the time-point of 10th week post-dose for rats administrated with PE section of EPR. (□) HCG and (△) PEG.

Figure 4. The comparison of biomarker intensity between HCG and PEG. (A) Hypoxanthine, (B) LPC C 20:4, (C) niacinamide, (D) phenylalanine, (E) C₁₆ dihydrosphingosine, (F) C₁₈ phytosphingosine, (G) N,N-dimethylglycine, (H) unknown, (I) unknown and (J) unknown. #Compared with HCG (p < 0.05) and *compared with HCG (p < 0.01).

Table 1. Potential kidney biomarkers of nephrotoxicity induced by EPR.

Retention time (min)	m/z (Da)	Scan mode	Quasi-molecular ion	Metabolite	Deviation ofnormal plasma
0.67	137	+	[M + H]^+	Hypoxanthine	↓
6.61	544	+	[M + H]^+	LPC C 20:4	↓
0.67	123	+	[M + H]^+	Niacinamide	↓
0.54	120	+	[M + H–HCOOH]^+	Phenylalanine	↑
0.67	274	+	[M + H]^+	C16 dihydrosphingosine	↑
0.67	318	+	[M + H]^+	C18 phytosphingosine	↑
0.55	104	+	[M + H]^+	N,N-Dimethylglycine	↑
0.63	118	+	Unknown	Unknown	↑
5.01	366	+	Unknown	Unknown	↓
5.46	368	+	Unknown	Unknown	↓

Figure 5. A set of chromatograms of (A) petroleum ether extract of EPR in positive ion mode, (B) petroleum ether extract of EPR in negative ion mode, (C) blank rat serum in positive ion mode, (D) blank rat serum in negative ion mode, (E) 1 h rat serum sample after oral administration of petroleum ether extract of EPR in positive ion mode and (F) 1 h rat serum sample after oral administration of petroleum ether section of EPR in negative ion mode.

Figure 6. Product ion scan spectra in negative ion mode under different collision energies (10, 20 and 25 eV) and possible MS fragmentation mechanism of (–)-(1S)-15-hydroxy-18-carboxycembrene.

Figure 7. Chemical structures of compounds identified from PE section.

Table 2. MS data (m/z) of the compounds of PE section of EPR and in rat serum after oral administration of PE section of EPR.

Peak no.	tR (min)	Quasi-molecularion	Formula	Measuredmass (m/z)	Calculatedmass (m/z)	mDa	Compound
1	10.8	[M + Na]^+	C27H22Na1O18	657.0709	657.0698	–1.04	Corilagin
2	11.3	[M + H]^+	C13H9O8	293.0292	293.0292	–0.01	Brevifolin carboxylic acid
		[M–H]^–	C13H7O8	291.0204	291.0146	–5.74
3	14.1	[M + H]^+	C12H9O6	249.0393	249.0394	0.07	Brevifolin
		[M–H]^–	C12H7O6	247.0273	247.0248	–2.52
4^a	15.2	[M + H]^+	C14H7O8	303.0162	303.0135	–2.68	Ellagic acid
		[M–H]^–	C14H5O8	301.0008	300.9990	–1.81
5^a	15.5	[M + H]^+	C22H21O13	493.0920	493.0977	5.66	3,3′-Di-O-methyl ellagic acid-4′-O-β-d-glucopyranoside
		[M–H]^–	C22H19O13	491.0861	491.0831	–2.99
6^a	18.1	[M + H]^+	C21H19O12	463.0903	463.0871	–3.16	3,3′-Di-O-methyl ellagic acid-4′-O-β-d-xylopyranoside
		[M–H]^–	C21H17O12	461.0730	461.0725	–0.47
7	19.8	[M + H]^+	C16H23O4	279.1581	279.1591	0.95	Dibutyl phthalate
8^a	22.9	[M + H]^+	C15H11O7	303.0500	303.0499	–0.10	Quercetin
		[M–H]^–	C15H9O7	301.0340	301.0354	1.39
9^a	23.4	[M + H]^+	C16H11O8	331.0448	331.0448	0.04	3,3′-Di-O-methyl ellagic acid
		[M–H]^–	C16H9O8	329.0304	329.0303	–0.08
10	27.7	[M + H]^+	C20H29O3	317.2091	317.2111	2.02	ent-(5β,8α,9β,10α,11α,12α)-11-Hydroxyatis-16-ene-3,14-dione
11	27.9	[M + H]^+	C20H27O3	315.1936	315.1955	1.86	Helioscopinolide E
12^a	28.0	[M–H]^–	C20H29O3	317.2124	317.2122	–0.16	(–)-(1S)-15-Hydroxy-18-carboxycembrene
13^a	28.5	[M + H]^+	C20H29O3	317.2078	317.2111	3.33	Yuexiandajisu C
14	29.2	[M–H]^–	C20H25O4	329.1784	329.1758	–2.59	Helioscopinolide C
15	29.7	[M + Na]^+	C20H30Na1O2	325.2127	325.2138	1.13	Pekinenal
16	29.7	[M + H]^+	C20H29O3	317.2069	317.2111	4.22	Helioscopinolide B
17	29.8	[M + H]^+	C20H29O3	317.2063	317.2111	4.83	Helioscopinolide
18^a	30.2	[M + Na]^+	C20H30Na1O2	325.2120	325.2138	1.84	Pekinenins A
19^a	31.7	[M + H]^+	C20H31O2	303.2274	303.2319	4.47	Euphorpekone A
20^a	31.7	[M + H]^+	C20H31O2	303.2274	303.2319	4.47	Euphorpekone B
21	34.5	[M + H]^+	C30H49O2	441.3643	441.3727	8.44	27-Hydroxy-3-oxolup-12-ene
22	34.7	[M + H]^+	C30H49O2	441.3642	441.3727	8.55	28-Hydrozylup-20(29)-en-3-one
23	35.9	[M + H–H2O]^+	C30H49O	425.3657	425.3778	12.05	3β,25-Dihydroxylanosta-8,23-diene
24	36.3	[M + H–H2O]^+	C30H49O	425.3659	425.3778	11.91	(24S)-9,19-Cycloart-25-ene-3β,24-diolh
25	36.3	[M + H–H2O]^+	C30H49O	425.3659	425.3778	11.91	(24R)-9,19-Cycloart-25-ene-3β,24-diolh

^a Compounds in rat serum after oral administration of PE section of EPR.