Simultaneous measurement of tadehaginoside and its principal metabolite in rats by HPLC–MS/MS and its application in pharmacokinetics and tissue distribution study

Figures & data

Figure 1. Structural formula of tadehaginoside (A), p-hydroxycinnamic acid (B) and quercetin (C).

Figure 2. Typical chromatograms of different samples (A1) Tadehaginoside in blank plasma; (A2) Tadehaginoside in blank matrix (spleen); (A3) Plasma sample at 5 min after intravenous administration of tadehaginoside; (A4) Kidney sample at 30 min after intravenous administration of tadehaginoside; (B1) Blank plasma spiked with IS; (B2) Blank matrix (spleen) spiked with IS; (B3) Plasma sample at 5 min after intravenous administration spiked with tadehaginoside and IS; (C1) HYD in Blank plasma; (C2) Plasma sample at 5 min after intravenous administration spiked with HYD; (C3) Plasma sample at 5 min after intragastric administration 5 min spiked with HYD.

Table 1. Parameters of standard curves of tadehaginoside and HYD in the rats as determined by LC-MS/MS during method validation.

Sample	Matrix	Run	Slope (10^–4)	Intercept (10^–3)	r
Tadehaginoside	Plasma	1	8.78	2.51	0.9967
		2	8.65	5.25	0.9985
		3	8.8	4.36	0.9984
	Liver	1	7.68	−1.22	0.9984
		2	7.37	7.03	0.997
		3	7.58	4.36	0.9988
HYD	Kidney	1	7.11	4.18	0.9985
		2	7.17	1.42	0.9974
		3	7.06	6.36	0.9962
	Plasma	1	16.1	1.69	0.9983
		2	17.1	2.85	0.9978
		3	17.9	2.99	0.9992

Table 2. Accuracy, precision, matrix effect and recovery of the LC-MS/MS method to determined tadehaginoside and HYD in rat plasma and various tissues (n = 6).

Sample	Matrix	concentration (ng/mL)	Batch	Within-run		Between-run		Matrix effect		Recovery
				Precision (RSD%)	Accuracy (% nominal)	Precision (RSD%)	Accuracy (% nominal)	Mean (%)	RSD (%)	Mean (%)	RSD (%)
Tadehaginoside	Plasma	1500	1	7.7	98	7.1	95.5	104.4	4.	79.9	7.8
			2	7.3	93.5
			3	6.7	94.9
		120	1	5.7	97.1	5.9	96.9	107.1	7.5	81.6	9.4
			2	6.9	97.3
			3	6.1	96.3
		3	1	8.2	96	9.1	98	108.4	10.6	80.2	4.3
			2	9.7	97.4
			3	10.1	100.7
	Liver	1500	1	7.4	97	8.8	99.8	95.3	1.2	116.0	3.9
			2	9.8	98.2
			3	8.7	104.3
		120	1	8.3	100.6	7.7	100.1	93.8	4.5	112.4	3.6
			2	8.7	98.2
			3	7.2	101.5
		12	1	9.5	109.2	8.5	107.2	104.7	7.4	104.8	8.6
			2	7.4	107.8
			3	9.6	104.7
	Kidney	1500	1	9.5	98.3	9.6	95.4	109.1	10.2	109.6	14.9
			2	9.1	91.6
			3	10.3	96.5
		120	1	6.4	98.2	6.6	98.2	100.8	13.1	106.7	5.6
			2	7.5	100.8
			3	6	95.6
		12	1	7.6	105.9	7.4	105.1	107.7	6.5	104.2	3.8
			2	8.6	103.5
			3	7.3	105.7
HYD	Plasma	1500	1	10.7	98.4	10.6	99.4	105.3	11.6	88.1	2.6
			2	11.0	100.6
			3	11.8	99.1
		120	1	7.4	98.6	8.1	98.94	103.4	5.7	84.6	9.6
			2	9.4	99.0
			3	8.9	99.2
		30	1	10.5	95.2	9.1	98.1	104.9	9.5	82.5	5.8
			2	9.0	98.9
			3	8.7	100.1

Table 3. Stability of the tadehaginoside and HYD in rat plasma and various tissues under different storage conditions.

Sample	Matrix	Concentration (ng/mL)	Room temperature				Auto-sampler 6 h		Stored at 4 °C 24 h		Three freeze-thraw cycles		Storage at −20 °C 7 days
			4 h		2h
			Recovery (%)	RSD (%)	Recovery (%)	RSD (%)	Recovery (%)	RSD (%)	Recovery (%)	RSD (%)	Recovery (%)	RSD (%)	Recovery (%)	RSD (%)
Tadehaginoside	Plasma	1500	95.1	4.2	113.6	2.5	104.4	3.8	115.2	1.7	105.6	4.5	95.7	11.5
		120	107.0	9.4	99.2	6.9	96.1	6.6	100.8	9.1	97.2	6.5	94.4	5.4
		3	80.5	0.2	96.6	4.0	104.0	7.5	94.0	4.0	94.1	4.4	82.3	7.4
	Kidney	1500	101.5	4.8	106.0	4.6	105.3	7.3	103.1	6.4	103.6	7.1	96.6	5.7
		120	100.7	5.8	96.4	4.1	95.0	2.6	95.8	3.8	97.2	10.6	92.8	11.5
		12	85.7	9.9	81.2	10.3	89.8	12.7	87.8	5.5	92.0	10.2	87.7	13.3
	Liver	1500	102.4	5.2	104.9	4.4	104.3	9.4	110.4	5.2	103.4	4.3	99.8	3.9
		120	98.7	5.5	93.3	3.6	96.4	6.9	97.8	9.9	102.8	13.0	95.0	9.4
		12	89.9	14.2	88.7	15.3	90.7	3.2	89.1	11.0	90.2	10.0	88.6	3.6
HYD	Plasma	1500	106.0	1.9	102.2	5.3	104.4	7.8	107.2	6.4	102.5	3.9	107.2	2.7
		120	99.7	6.0	94.2	9.2	97.5	7.0	100.3	4.1	98.6	13.9	96.5	7.1
		30	86.0	6.2	85.6	6.0	84.1	1.5	84.6	5.1	83.3	4.0	80.9	2.6

Figure 3. Mean plasma concentration–time curves of tadehaginoside and p-hydroxycinnamic acid after (A), (C) intragastric administration (25 mg/kg); (B), (D) intravenous administration (5 mg/kg) to rats.

Figure 4. Concentration of tadehaginoside in rat tissues determined by HPLC-MS/MS.

Table 4. Pharmacokinetic parameters of tadehaginoside and HYD after intragastric and intravenous administrations.

PK parameters	i.g. (25 mg/kg)		i.v. (5 mg/kg)
	Tadehaginoside	HYD	Tadehaginoside	HYD
t1/2 (h)	2.51 ± 2.21	1.24 ± 0.038	1.27 ± 1.19	0.86 ± 0.58
Cmax (ng/mL)	6.01 ± 2.15	837.75 ± 446.66	109.77 ± 4.29	1536.45 ± 193.93
Tmax (h)	0.25 ± 0.08	1.25 ± 0.43	0.08 ± 0.00	0.08 ± 0.00
AUC0–t (h ng/mL)	7.92 ± 5.43	2027.58 ± 1091.20	52.85 ± 5.11	597.24 ± 103.90
AUC0–∞ (h ng/mL)	14.12 ± 4.60	2099.87 ± 1103.33	59.22 ± 10.79	608.43 ± 98.42
MRT (0–t) (h)	0.52 ± 0.44	1.84 ± 0.11	0.54 ± 0.07	0.39 ± 0.07
Vz (L/kg)	6089.77 ± 4011.02	30.63 ± 28.50	143.27 ± 102.37	10.96 ± 8.95
CLz (L/h/kg)	1937.91 ± 770.51	15.19 ± 9.74	86.44 ± 13.88	8.38 ± 1.28

Figure 5. The mass spectrum of tadehaginoside [(A), Q1-scan; (B) Full-scan], p-hydroxycinnamic acid [(C), Q1-scan; (D) Full-scan] and quercetin [(E), Q1-scan; (F) Full-scan].