Study on the pharmacokinetics, tissue distribution and excretion of laurolitsine from Litsea glutinosa in Sprague-Dawley rats

Figures & data

Figure 1. Chemical structures and MS/MS spectra of laurolitsine and nuciferine.

Figure 2. LC-MS/MS chromatograms oflaurolitsine and IS nuciferine in different matrix. (A) Blank biological matrix; (B) LLOQ sample of different biological matrix (laurolistine 1 ng/mL⁻¹); (C) rat plasma of 1 h after i.v. administration laurolistine, rat urine of 0–4 h after p.o. administration laurolistine, rat liver tissue of 2 h after p.o. administration laurolistine; (D) rat plasma of 1 h after p.o. administration laurolitsine.

Table 1. Standard calibration curves and LLOQ of laurolitsine in different matrix.

Matrix	Equation (1/x^2, Weighting Index)	Range (ng/mL)	LLOQ (ng/mL)
Plasma	y = 0.00741x + 0.00319 (r = 0.9979)	1–2000	1
Liver	y = 0.01438x − 0.00754 (r = 0.99885)	1–2000	1
Kidney	y = 0.01238x − 0.00283 (r = 0.99935)	1–2000	1
Spleen	y = 0.01566x + 0.00106 (r = 0.99968)	1–2000	1
Lung	y = 0.01359x − 0.00366 (r = 0.99938)	1–2000	1
Brain	y = 0.01704x − 0.00697 (r = 0.99804)	1–2000	1
Heart	y = 0.01385x + 0.00144 (r = 0.99583)	1–2000	1
Testis	y = 0.01286x − 0.00393 (r = 0.99929)	1–2000	1
Stomach	y = 0.01060x + 0.00292 (r = 0.99812)	5–10000	5
Muscle	y = 0.01717x − 0.00045 (r = 0.99838)	1–2000	1
Fat	y = 0.01771x − 0.00126 (r = 0.99944)	1–2000	1
Intestine	y = 0.01258x − 0.00023 (r = 0.99687)	1–2000	1
Urine	y = 0.0122x − 0.01301 (r = 0.9984)	1–2000	1
Faeces	y = 0.0153x + 0.0203 (r = 0.9994)	5–10000	5

Table 2. Intra-day and inter-day precision, accuracy of laurolitsine.

Matrix	Laurolitsine Spiked (ng/mL^−1)	Intra-day (n = 6)		Accuracy (%)	Inter-day (n = 18)		Accuracy (%)
		Measured (ng/mL)	RSD (%)		Measured (ng/mL)	RSD (%)
Plasma	3	3.30 ± 0.21	6.25	110.2	3.18 ± 0.24	7.45	105.88
	120	118.24 ± 5.51	4.66	98.54	118.49 ± 6.55	5.53	98.74
	1500	1513.38 ± 97.21	6.42	100.89	1495.96 ± 90.69	6.06	99.73
Liver	3	2.92 ± 0.25	8.62	97.30	2.97 ± 0.23	7.61	99.07
	150	161.03 ± 6.55	4.07	107.36	156.64 ± 8.42	5.38	104.43
	1500	1445.28 ± 84.33	5.83	96.35	1505.16 ± 111.74	7.42	100.34
Kidney	3	2.93 ± 0.28	9.72	97.65	3.02 ± 0.29	9.57	100.59
	150	153.88 ± 9.91	6.44	102.59	154.79 ± 8.86	5.73	103.20
	1500	1429.94 ± 72.77	5.09	95.33	1485.20 ± 96.66	6.51	99.01
Urine	3	3.21 ± 0.22	6.93	106.83	3.18 ± 0.23	7.10	105.98
	150	149.95 ± 12.17	8.11	99.97	151.80 ± 8.97	5.91	101.20
	1500	1508.78 ± 98.85	6.55	100.59	1527.17 ± 84.28	5.52	101.81
Faeces	12	11.78 ± 0.97	8.26	96.10	11.89 ± 0.94	7.92	99.11
	750	723.19 ± 54.12	7.48	90.95	728.39 ± 51.10	7.02	97.12
	7500	7465.22 ± 315.39	4.22	98.13	7470.99 ± 301.06	4.03	99.61

Table 3. Stability of laurolitsine in plasma, liver, kidney, urine, faeces (n = 6).

Matrix	Investigation conditions	Theoretical concentration (ng/mL)	Measured concentration (Mean ± SD) (ng/mL)	RSD (%)	Accuracy (%)
Plasma	Room temperature stability (25 °C, 6 h)	3.00	2.99 ± 0.22	7.47	99.67
		120.00	117.90 ± 6.87	5.83	98.25
		1500.00	1591.77 ± 90.47	5.68	106.12
	Short time stability (4 °C, 24 h)	3.00	3.04 ± 0.14	4.48	101.33
		120.00	119.31 ± 5.60	4.70	99.43
		1500.00	1542.88 ± 56.57	3.67	102.86
	Freeze thaw stability (12 h/12 h)	3.00	3.10 ± 0.24	7.63	103.33
		120.00	126.27 ± 5.32	4.21	105.23
		1500.00	1404.27 ± 62.12	4.42	93.62
	Long time freezing stability (−20 °C, 7 d)	3.00	3.07 ± 0.26	8.35	102.33
		120.00	121.55 ± 8.67	7.12	101.29
		1500.00	1456.63 ± 61.21	4.20	97.11
Liver	Room temperature stability (25 °C, 6 h)	3.00	3.06 ± 0.14	4.65	102.00
		150.00	150.43 ± 9.86	6.55	100.29
		1500.00	1526.32 ± 86.93	5.70	101.75
	Short time stability (4 °C, 24 h)	3.00	2.97 ± 0.29	9.65	99.00
		150.00	152.75 ± 10.95	7.17	101.83
		1500.00	1519.98 ± 110.13	7.25	101.33
	Freeze thaw stability (12h/12 h)	3.00	3.15 ± 0.24	7.69	105.00
		150.00	149.51 ± 15.07	10.08	99.67
		1500.00	1519.96 ± 151.86	9.99	101.33
	Long time freezing stability (−20 °C, 7 d)	3.00	3.03 ± 0.28	9.34	101.00
		150.00	147.36 ± 15.47	10.50	98.24
		1500.00	1554.67 ± 122.62	7.89	103.64
Kidney	Room temperature stability (25 °C, 6 h)	3.00	3.05 ± 0.13	4.39	101.67
		150.00	149.70 ± 7.28	4.87	99.80
		1500.00	1515.43 ± 5.84	5.84	101.03
	Short time stability (4 °C, 24 h)	3.00	2.97 ± 0.26	8.90	99.00
		150.00	147.70 ± 11.28	7.63	98.47
		1500.00	1476.49 ± 78.73	5.33	98.43
	Freeze thaw stability (12h/12 h)	3.00	2.97 ± 0.27	8.91	99.00
		150.00	151.00 ± 11.41	7.56	100.67
		1500.00	1475.38 ± 142.92	9.69	98.36
	Long time freezing stability (−20 °C, 7 d)	3.00	3.06 ± 0.28	7.65	102.00
		150.00	145.12 ± 16.62	11.45	96.75
		1500.00	1493.25 ± 145.80	9.76	99.55
Urine	Room temperature stability (25 °C, 6 h)	3.00	3.03 ± 0.10	3.35	101.00
		150.00	148.71 ± 6.97	4.69	99.14
		1500.00	1509.65 ± 77.34	5.12	100.64
	Short time stability (4 °C, 24 h)	3.00	3.02 ± 0.28	9.34	100.67
		150.00	149.58 ± 7.27	4.86	99.72
		1500.00	1472.03 ± 111.54	7.58	98.14
	Freeze thaw stability (12 h/12 h)	3.00	3.02 ± 0.32	10.62	100.67
		150.00	151.50 ± 13.45	8.88	101.00
		1500.00	1477.47 ± 114.99	7.78	98.50
	Long time freezing stability (−20 °C, 7 d)	3.00	3.08 ± 0.20	6.50	102.67
		150.00	148.02 ± 13.57	9.17	98.68
		1500.00	1522.16 ± 91.75	6.03	101.48
Faeces	Room temperature stability (25 °C, 6 h)	12.00	3.05 ± 0.13	6.14	25.42
		750.00	149.70 ± 7.28	5.24	19.96
		7500.00	1515.43 ± 5.84	3.36	20.21
	Short time stability (4 °C, 24 h)	12.00	2.97 ± 0.26	11.37	24.75
		750.00	147.70 ± 11.28	4.21	19.69
		7500.00	1476.49 ± 78.73	4.22	19.69
	Freeze thaw stability (12 h/12 h)	12.00	2.97 ± 0.27	9.18	24.75
		750.00	151.00 ± 11.41	6.23	20.13
		7500.00	1475.38 ± 142.92	3.76	19.67
	Long time freezing stability (−20 °C, 7 d)	12.00	3.06 ± 0.28	8.24	25.50
		750.00	145.12 ± 16.62	8.22	19.35
		7500.00	1493.25 ± 145.80	4.65	19.91

Table 4. Extraction recovery and matrix effect of laurolitsine (Mean ± SD, n = 6).

Matrix	Concentration (ng/mL)	Extraction recovery (%)	Matrix effect (%)
Plasma	3.00	89.00 ± 4.52	91.99 ± 11.10
	120.00	92.51 ± 6.52	93.67 ± 2.63
	1500.00	92.84 ± 10.41	95.94 ± 9.23
Liver	3.00	89.74 ± 4.01	97.87 ± 9.33
	150.00	94.39 ± 8.33	96.33 ± 4.81
	1500.00	91.22 ± 6.87	96.87 ± 7.95
Kidney	3.00	87.37 ± 5.78	101.14 ± 6.83
	150.00	90.24 ± 6.75	93.01 ± 5.90
	1500.00	95.21 ± 5.85	96.92 ± 7.80
Urine	3.00	91.68 ± 5.22	95.82 ± 8.09
	150.00	94.13 ± 6.34	96.92 ± 4.86
	1500.00	93.05 ± 6.53	101.69 ± 4.09
Faeces	12.00	93.25 ± 7.97	103.21 ± 9.71
	750.00	87.13 ± 8.32	98.97 ± 6.51
	7500.00	93.05 ± 5.75	97.54 ± 7.16

Figure 3. Plasma concentration-time curve of laurolitsine after i.g. (10 mg/kg) and i.v. (2 mg/kg) administration to SD rats (n = 5).

Table 5. Pharmacokinetic parameters of laurolitsine after i.g. and i.v. admininstration.

PK parameters	i.v. Admininstration (2 mg/kg)	i.g. Admininstration (10 mg/kg)
t1/2 (h)	1.67 ± 1.83	3.73 ± 2.48
Cmax (ng/mL)	100.05 ± 17.00	14.11 ± 3.30
Tmax(h)	0.083	0.47 ± 0.18
AUC0-t (h·ng/mL)	49.98 ± 7.39	40.62 ± 9.57
AUC0-∞ (h·ng/mL)	52.29 ± 7.98	47.77 ± 10.44
MRT (0-t)(h)	0.61 ± 0.22	2.90 ± 0.53
Vz(L/kg)	86.04 ± 86.75	1207.59 ± 799.48
CLz (L/h/kg)	38.96 ± 5.86	218.26 ± 51.89
F(%)	–	18.17

Figure 4. The mean tissue drug concentrations at 0.5, 1, 2, 4, and 6 h after the oral administration of 10 mg/kg laurolitsine.

Figure 5. The cumulative excretion of laurolitsine in the urine and faeces of rats after oral administration 10 mg/kg as a percentage of the dosage and excretion rate.