Comparative pharmacokinetic analysis of raw and steamed Panax notoginseng roots in rats by UPLC-MS/MS for simultaneously quantifying seven saponins

Figures & data

Figure 1. Chemical structures and product ion mass spectra of GRg₁ (A), GRe (B), GRb₁ (C), GRd (D), GRg₅ (E), GRk₁ (F) and NGR₁ (G).

Figure 2. Representative MRM chromatograms of GRb₁ (1), GRe (2), GRd (3), NGR₁ (4), GRg₁ (5), GRg₅ (6), GRk₁ (7) and theophylline (I.S., 8) in rat plasma: (A) blank rat plasma; (B) blank rat plasma spiked with standard compounds and IS; (C) plasma samples after oral administration of Panax notoginseng aqueous extract.

Table 1. Accuracy and precision of seven compounds from QC samples in rat plasma (n = 3 d, 6 replicates per day).

Compounds	Spiked concentration (ng/mL)	Intra-day data		Inter-day data
		Accuracy (RE%)	Precision (RSD%)	Accuracy (RE%)	Precision (RSD%)
Rg1	2.39	−11.2	7.62	−3.26	5.24
	47.9	3.82	8.49	−0.37	5.14
	95.6	1.29	2.39	−1.53	3.35
Re	1.20	−4.83	13.4	−0.67	12.1
	23.9	0.22	11.8	4.37	8.32
	47.9	−3.42	5.00	1.29	3.99
NGR1	2.39	−0.59	15.1	10.0	7.56
	47.9	4.94	4.95	−5.30	3.78
	95.6	−7.70	1.75	−2.28	2.37
Rd	9.56	8.33	11.5	2.64	10.5
	95.6	7.01	5.74	5.65	5.25
	191.3	−2.96	3.28	−1.53	4.06
Rb1	7.65	1.23	11.1	−4.71	7.30
	765.1	7.87	5.88	1.72	5.08
	1530.2	−3.91	6.40	−6.03	8.05
Rg5	19.1	9.42	6.08	10.9	3.74
	95.6	0.31	2.76	4.02	2.02
	191.3	−0.90	8.37	−4.20	4.50
Rk1	19.1	−8.90	7.31	−3.35	7.70
	95.6	−10.8	4.36	5.69	3.54
	191.3	6.43	2.65	8.71	5.21

Table 2. Matrix effects and extraction recoveries of seven compounds in rat plasma (n = 5).

Compounds	Spiked concentration (ng/mL)	Matrix effect (%)		Extraction recovery (%)
		Mean (%)	RSD (%)	Mean (%)	RSD (%)
Rg1	2.39	89.3	10.3	98.8	8.64
	95.6	105.2	9.04	94.8	4.49
Re	1.20	97.3	10.6	88.0	7.44
	47.9	110.0	7.09	100.3	3.31
NGR1	2.39	96.9	9.06	86.8	2.80
	95.6	110.3	8.53	100.4	3.25
Rd	9.56	90.4	12.2	100.8	8.36
	191.3	103.4	11.3	89.3	5.53
Rb1	7.65	94.7	10.3	101.2	5.72
	1530.2	100.7	8.33	103.2	6.58
Rg5	19.1	89.3	5.31	76.8	4.83
	191.3	85.1	7.52	75.0	3.47
Rk1	19.1	87.7	9.19	77.9	4.42
	191.3	85.6	6.57	82.0	3.36

Table 3. Stability data of seven compounds and IS in rat plasma under various storage conditions (n = 3).

Compounds	Spiked concentration (ng/mL)	Ambient temperature stability		Long-term stability		Three freeze/thaw cycles		Post-preparative stability
		Accuracy (RE%)	Precision (RSD%)	Accuracy (RE%)	Precision (RSD%)	Accuracy (RE%)	Precision (RSD%)	Accuracy (RE%)	Precision (RSD%)
Rg1	2.39	−5.07	6.9	3.52	9.6	−0.13	6.50	5.72	10.5
	95.6	2.86	2.07	−2.33	5.5	2.26	1.38	5.9	4.4
Re	1.20	−0.83	14.6	7.21	15.0.	−6.71	3.0	6.2	12.1
	47.9	−0.53	5.40	5.83	8.00	1.33	5. 5	5.44	3.6
NGR1	2.39	−4.35	3.4	2.05	5.93	−7.43	11.1	4.64	5.1
	95.6	−0.77	3.13	−9.83	9.24	6.54	6.43	2.3	3.4
Rd	9.56	5.26	8.6	6.45	10.7	−3.96	10.9	−4.23	13.8
	191.3	2.75	7.7	1.30	0.74	7.37	5.25	2.51	4.03
Rb1	7.65	−1.63	6.94	−4.70	11.23	0.54	5.81	1.85	3.45
	1530.2	−4.93	8.28	4.35	10.94	−3.46	7.14	−1.42	5.52
Rg5	19.1	2.92	6.42	7.46	8.12	4.62	9.53	10.6	3.36
	191.3	3.48	6.24	6.28	5.99	5.23	10.2	0.25	1.29
Rk1	19.1	5.03	5.18	3.25	8.36	7.25	8.66	3.09	8.43
	191.3	1.35	3.95	7.38	1.52	0.95	8.33	5.15	2.86
IS	2.39	6.78	1.18	−9.62	2.17	1.99	2.02	−8.93	6.58

Figure 3. Saponin content in steaming Panax notoginseng root. The label on the left was for GRb₁, GRe, GRd, NGR₁ and GRg₁ and the label on the right was for GRg₅ and GRk₁.

Figure 4. Mean plasma concentration–time profiles of NGR₁ (A), GRb₁ (B), GRg₁ (C), GRd (D) and GRe (E) in the Sprague–Dawley rat after oral administration the extracts of Panax notoginseng at 5 mL/kg. Each point represents the means ± SD.

Table 4. Pharmacokinetic parameters for GRg₁, GRe, NGR₁, GRd and GRb₁ in male SD rats following oral gavage of raw and steam-processed Panax notoginseng aqueous extracts (n = 6, mean ± SD).

Analyte	Groups	Cmax (ng/mL)	Tmax (h)	AUC0–t (ng·h/mL)	AUC0–∞ (ng·h/mL)	t1/2 (h)
Rg1	Raw	66.67 ± 32.81	9.33 ± 2.07	808.42 ± 382.14	860.90 ± 354.81	7.92 ± 6.73
	Steam-processed	17.41 ± 5.43**	8.67 ± 1.63	176.63 ± 42.49**	194.85 ± 46.90**	7.46 ± 2.47
NGR1	Raw	94.31 ± 55.29	1.12 ± 0.33	578.05 ± 273.25	627.83 ± 303.61	7.63 ± 0.38
	Steam-processed	23.97 ± 16.77*	1.25 ± 0.00	135.95 ± 54.32**	156.52 ± 57.94**	9.27 ± 2.01
Re	Raw	28.15 ± 13.10	1.24 ± 0.49	87.71 ± 20.68	91.69 ± 19.68	3.16 ± 1.73
	Steam-processed	21.53 ± 10.02	1.63 ± 0.41	102.08 ± 24.69	106.50 ± 24.22	4.14 ± 1.98
Rb1	Raw	997.00 ± 143.69	8.67 ± 1.63	15110.08 ± 2383.47	15242.07 ± 2435.24	9.58 ± 1.11
	Steam-processed	361.48 ± 165.57***	8.33 ± 1.97	5094.06 ± 1453.14***	5446.36 ± 1661.83***	9.91 ± 0.83
Rd	Raw	164.37 ± 28.80	8.67 ± 2.73	3633.64 ± 568.69	4172.87 ± 733.16	13.95 ± 1.71
	Steam-processed	62.47 ± 33.65***	8.33 ± 1.97	1396.89 ± 595.14***	1538.71 ± 594.79***	12.01 ± 3.06

C_max: maximum concentration; T_max: time to maximum concentration; AUC_0–t: area under the concentration–time curve from zero to last sampling time; AUC_0–∞, area under the concentration–time curve from zero to infinity; t_1/2: elimination half-life.

*Indicated p < 0.05, **indicated p < 0.01 and ***indicated p < 0.001, versus oral administration of raw Panax notoginseng aqueous extract.