Plasma metabolic profiling of normal and dysmenorrhea syndrome rats and the effects of Xiang–Fu–Si–Wu Decoction intervention

Figures & data

Table 1. Effects of XFSWD on the level of writhing scores, Ca²⁺, and nitric oxide (NO).

Group	Writhing scores	NO (μmol/gprot)	Ca^2+(mmol/gprot)
A	1.2 ± 0.8	25.79 ± 4.35	1.32 ± 0.65
B	49.6 ± 3.5***	15.86 ± 4.07**	2.29 ± 0.84*
C	2.2 ± 1.3	27.60 ± 8.13	1.25 ± 0.77
D	16.8 ± 3.7†††	21.38 ± 5.35††	1.88 ± 0.54†

n = 5. ***p < 0.001, **p < 0.01, *p < 0.05 versus group A. †††p < 0.001, ††p < 0.01, †p < 0.05 versus group B.

Figure 1. Representative UHPLC-Q-TOF-MS base peak intensity (BPI) chromatogram of a rat plasma sample analyzed in negative and positive ion modes.

Table 2. The variations of retention times and m/z values and the relative standard deviations of peaks areas from reference compound peaks in positive and negative ESI modes (n = 12).

	Positive mode			Negative mode
Reference compound	RT	m/z	Peak area (%)	RT	m/z	Peak area (%)
Caffeic acid	–	–	--	0.007	0.001	3.50
Ferulaic acid	–	–	--	0.012	0.001	2.24
Tetrahydropalmatine	0.008	0.013	1.69	–	–	–
Berberine	0.005	0.008	1.99	–	–	–

Figure 2. PCA-DA scores plots. (a) Negative ion mode, (b) positive ion mode, (▪) normal group without treatment, and (▴) dysmenorrhea syndrome model group without treatment.

Figure 3. PLS-DA scores plots. (a) Negative ion mode, (b) positive ion mode, (▪) normal group without treatment, and (▴) dysmenorrhea syndrome model group without treatment.

Figure 4. OPLS-DA S-plot. (a) Negative ion mode and (b) positive ion mode.

Table 3. Potential markers from rats with dysmenorrhea syndrome (p < 0.05) and their identification results on ESI⁺ and ESI⁻ scans.

No.	ESI ionization mode	tR (min)	Experimental m/z	Mass accuracy^a	Δppm	Formula	Fragment ion on MSE	Proposed compound	Trend (model versus normal)^b	p Value (t test)
1	ESI^−	4.15	514.2934	515.3012	0	C26H46NO7P	498, 467, 369, 271	LysoPC(18:4)^C	↓	8.64E−3
2	ESI^−	6.33	578.3483	533.3481	4.3	C27H52NO7P	504, 271	LysoPE(22:2/0:0)	↓	2.41E−4
3	ESI^−	2.85	417.1183	418.1264	0.7	C21H22O9	343, 325, 279	Unidentified	↓	1.66E−3
4	ESI^−	6.48	554.3475	509.3481	2.2	C25H52NO7P	480, 271	LysoPC(17:0)	↓	8.50E−5
5	ESI^−	4.61	333.2071	334.2144	1.5	C20H30O4	249, 217, 174	PGJ2	↑	2.37E−3
6	ESI^−	5.76	317.2131	318.2195	4.4	C20H30O3	255, 217, 157	Unidentified	↑	3.34E−2
7	ESI^−	4.73	349.2384	350.2457	1.4	C21H34O4	271, 255, 217	11-Deoxy- 11-methylene-PGD2	↑	1.86E−2
8	ESI^−	5.35	315.1942	316.2038	5.7	C20H28O3	271, 255	15-Deoxy-δ-12, 14-PGJ2	↑	3.40E−4
9	ESI^+	6.36	521.3148	520.3036	6.5	C29H44O8	502, 312, 184	Unidentified	↓	2.11E−4
10	ESI^+	6.83	510.3385	545.3481	7.2	C28H52NO7P	349, 305, 261	LysoPC(20:3)	↓	1.98E−4
11	ESI^+	6.23	392.2013	391.1968	8.4	C17H25N7O4	364, 228, 217	Unidentified	↑	5.04E−4
12	ESI^+	3.98	247.2406	246.2371	8.1	C18H30	283, 228, 219	Unidentified	↑	9.17E−4

^aMass accuracy (ppm) was calculated according to the exact mass on the Pubchem website.

^bCompared with normal rats. (↑) Up-regulated. (↓) Down-regulated.

^cConfirmed by standard samples.

Figure 5. Identification of a selected marker. (A) Peak of potential biomarker of m/z 315 in extracted ion chromatogram with negative mode. (B) Corresponding mass spectrum. (C) MS^E spectrum, the collision energy was 25 eV.

Figure 6. Mean peak area (mean±standard error) of markers (a–l) from normal and dysmenorrhea syndrome rats plasma samples after administration of XFSWD at 30 min, 2 h, 6 h, and 12 h (n = 5). (▪) Dysmenorrhea syndrome model group with treatment and (□) normal group with treatment.

Figure 7. Scheme illustrating sphingolipid metabolism and Xiang–Fu–Si–Wu Decoction intervention.