Pharmacokinetic analysis and tissue distribution of andrographolide in rat by a validated LC-MS/MS method

Figures & data

Figure 1. (a) Q1 and product ion scan data of AG in LC-MS/MS and (b) Q1 and product ion scan data of diclofenac in LC-MS/MS.

Table 1. MS parameters of AG and diclofenac.

Compound	Q1	Q3	DP	EP	FP	CE	CXP	CAD	IS
AG	349.20	287.20	−40	−10	100	−20	−12	5	−4500
Diclofenac	294.10	213.90	−50	−10	100	−30	−12	5	−4500

Figure 2. (a) Representative chromatogram of blank plasma, (b) representative chromatogram of blank kidney and (c) representative chromatogram of unknown plasma sample with analyte and IS.

Table 2. Intra-day and inter-day accuracy and precision of AG.

	Intra-day accuracy and precision				Inter-day accuracy and precision
	Quality	Concentration of AG (ng/ml)
Matrix	control	Mean	% CV	% Accuracy	Mean	% CV	% Accuracy
Plasma	LLOQ	4.05	6.67	103.58	3.89	4.11	99.49
	LQC	11.00	4.59	91.67	10.72	3.92	89.33
	MQC	197.18	4.23	98.59	212.25	3.10	106.13
	HQC	794.19	1.86	99.27	815.42	2.07	101.93
Whole blood	LLOQ	3.95	4.81	101.02	4.15	5.06	106.14
	LQC	11.25	2.84	93.75	10.65	2.75	93.83
	MQC	195.72	1.86	97.86	200.48	5.28	100.24
	HQC	799.56	1.28	99.95	826.48	2.01	103.31
Liver	LLOQ	3.86	3.89	98.72	4.12	4.61	105.37
	LQC	12.03	2.99	100.25	11.67	1.80	97.25
	MQC	210.25	4.92	105.13	208.51	6.96	104.26
	HQC	825.52	2.57	103.19	812.52	2.04	101.57
Lung	LLOQ	4.02	2.99	102.81	3.85	2.08	98.47
	LQC	11.86	1.60	98.83	12.85	8.72	107.08
	MQC	198.62	4.36	99.31	188.64	6.65	94.32
	HQC	805.23	3.56	100.65	812.57	2.65	101.57
Kidney	LLOQ	3.95	3.54	101.02	4.24	2.59	108.44
	LQC	12.16	3.50	101.33	11.32	4.77	94.33
	MQC	195.58	6.15	97.79	200.89	5.81	100.45
	HQC	821.25	4.29	102.66	835.69	4.26	104.46
Brain	LLOQ	4.12	5.34	105.37	3.88	4.38	99.23
	LQC	11.56	1.99	96.33	12.06	1.74	100.50
	MQC	185.58	4.66	92.79	196.59	6.39	98.30
	HQC	789.25	3.24	98.66	809.58	2.91	101.20
Testes	LLOQ	3.86	2.07	98.72	4.08	6.37	104.35
	LQC	12.12	2.15	101.00	11.34	7.85	94.50
	MQC	198.52	2.87	99.26	191.28	5.51	93.18
	HQC	789.59	3.25	98.70	763.58	4.70	95.45
Heart	LLOQ	4.22	1.42	107.93	3.52	3.98	90.03
	LQC	11.08	5.05	92.33	11.55	4.24	96.25
	MQC	215.56	4.76	107.78	206.38	4.69	103.19
	HQC	826.25	3.23	103.28	800.29	4.56	100.04
Spleen	LLOQ	3.82	3.93	97.70	4.08	7.11	104.35
	LQC	11.25	4.80	93.75	12.34	3.81	102.83
	MQC	212.34	7.38	106.17	208.49	7.04	104.25
	HQC	815.14	4.36	101.89	809.18	4.01	101.15

%CV, Coefficient of variation (SD × 100/mean). Accuracy, calculated concentration × 100/nominal concentration.

Table 3. Stability study data of AG.

	Stability test data of AG
			Measured concentration (ng/ml)
Matrix	QC	Stability	Mean	% CV	% Accuracy
Plasma	LQC	0 h	12.35	7.45	102.92
		24 h autosampler	10.96	5.11	91.33
		8 h bench-top	11.95	5.19	99.58
		Third freeze-thaw	11.02	8.71	91.83
		30 days at −20 °C	10.56	7.77	88.00
	HQC	0 h	805.8	3.10	100.73
		24 h autosampler	793.28	3.17	99.16
		8 h bench-top	787.58	3.19	98.45
		Third freeze-thaw	784.53	3.53	98.07
		30 days at −20 °C	723.78	3.61	90.47
Whole blood	LQC	0 h	12.74	3.53	106.17
		24 h autosampler	12.05	5.15	100.42
		8 h bench-top	10.98	3.92	91.50
		Third freeze-thaw	11.96	3.09	99.67
		30 days at −20 °C	10.87	5.34	90.58
	HQC	0 h	806.58	0.00	100.82
		24 h autosampler	789.91	3.87	98.74
		8 h bench-top	812.47	3.96	101.56
		Third freeze-thaw	782.43	3.65	97.80
		30 days at −20 °C	791.14	5.13	98.89
Liver	LQC	0 h	12.54	0.96	104.50
		24 h autosampler	11.45	2.97	95.42
		8 h bench-top	12.06	1.74	100.50
		Third freeze-thaw	11.14	5.75	92.83
		30 days at −20 °C	10.82	9.70	90.17
	HQC	0 h	766.36	5.94	95.80
		24 h autosampler	799.51	5.26	99.94
		8 h bench-top	800.64	7.94	100.08
		Third freeze-thaw	786.39	3.54	98.30
		30 days at −20 °C	802.24	7.05	100.28
Lung	LQC	0 h	12.03	2.66	100.25
		24 h autosampler	12.19	3.94	101.58
		8 h bench-top	11.92	6.29	99.33
		Third freeze-thaw	11.68	5.57	97.33
		30 days at −20 °C	11.76	4.00	98.00
	HQC	0 h	789.69	5.89	98.71
		24 h autosampler	785.29	3.83	98.16
		8 h bench-top	768.48	4.64	96.06
		Third freeze-thaw	813.34	3.15	101.67
		30 days at −20 °C	800.49	8.05	100.06
Kidney	LQC	0 h	12.08	2.65	100.67
		24 h autosampler	11.54	1.56	96.17
		8 h bench-top	13.57	1.62	113.08
		Third freeze-thaw	10.84	4.43	90.33
		30 days at −20 °C	11.08	3.25	92.33
	HQC	0 h	800.48	5.65	100.06
		24 h autosampler	810.25	1.52	101.28
		8 h bench-top	821.31	7.58	102.66
		Third freeze-thaw	792.34	5.34	99.04
		30 days at −20 °C	786.56	4.48	98.32
Brain	LQC	0 h	12.54	3.59	104.50
		24 h autosampler	12.03	2.00	100.25
		8 h bench-top	11.64	4.55	97.00
		Third freeze-thaw	10.76	2.42	89.67
		30 days at −20 °C	12.48	1.20	104.00
	HQC	0 h	756.65	2.81	94.58
		24 h autosampler	762.36	3.29	95.30
		8 h bench-top	797.48	6.84	99.69
		Third freeze-thaw	825.35	5.91	103.17
		30 days at −20 °C	863.61	4.54	107.95
Testes	LQC	0 h	12.24	2.61	102.00
		24 h autosampler	12.65	1.90	105.42
		8 h bench-top	11.43	4.20	95.25
		Third freeze-thaw	11.08	2.62	92.33
		30 days at −20 °C	10.62	3.20	88.50
	HQC	0 h	879.89	3.57	109.99
		24 h autosampler	854.32	5.34	106.79
		8 h bench-top	812.25	3.33	101.53
		Third freeze-thaw	795.64	4.02	99.46
		30 days at −20 °C	790.06	7.91	98.76
Heart	LQC	0 h	12.30	2.76	102.50
		24 h autosampler	12.14	3.46	101.17
		8 h bench-top	11.52	7.73	96.00
		Third freeze-thaw	10.78	5.66	89.83
		30 days at −20 °C	11.06	1.08	92.17
	HQC	0 h	841.08	2.81	105.14
		24 h autosampler	822.27	1.89	102.78
		8 h bench-top	811.12	1.51	101.39
		Third freeze-thaw	800.26	8.03	100.03
		30 days at −20 °C	831.48	2.80	103.94
Spleen	LQC	0 h	12.18	4.43	101.50
		24 h autosampler	12.07	1.99	100.58
		8 h bench-top	12.06	4.15	100.50
		Third freeze-thaw	11.09	2.07	92.42
		30 days at −20 °C	10.89	5.88	90.75
	HQC	0 h	862.35	2.84	107.79
		24 h autosampler	823.36	2.58	102.92
		8 h bench-top	798.89	7.93	99.86
		Third freeze-thaw	768.48	6.32	96.06
		30 days at −20 °C	800.12	5.30	100.02

Table 4. Extraction recovery and ME of AG.

	Spiked concentration (ng/ml)
	12		200		800
Matrix	Recovery (%)	ME (%)	Recovery (%)	ME (%)	Recovery (%)	ME (%)
Plasma	86.94 ± 5.65	3.81 ± 0.29	87.12 ± 3.91	3.02 ± 0.19	83.21 ± 2.03	3.21 ± 0.22
Whole blood	83.24 ± 2.98	3.81 ± 0.32	88.55 ± 4.22	2.78 ± 0.41	85.44 ± 4.52	2.98 ± 0.15
Liver	85.45 ± 3.53	2.44 ± 0.14	87.14 ± 2.98	3.07 ± 0.32	84.16 ± 3.44	3.14 ± 0.23
Lung	86.37 ± 2.56	2.78 ± 0.46	85.14 ± 4.02	4.37 ± 0.42	88.26 ± 4.24	2.27 ± 0.25
Kidney	82.47 ± 3.48	3.08 ± 0.24	84.74 ± 3.32	3.24 ± 0.31	86.45 ± 3.88	3.67 ± 0.42
Brain	89.35 ± 4.25	2.96 ± 0.32	88.54 ± 4.14	2.24 ± 0.21	87.55 ± 2.82	3.23 ± 0.34
Testes	88.51 ± 3.75	1.82 ± 0.27	90.15 ± 5.58	2.24 ± 0.11	88.41 ± 2.17	3.43 ± 0.14
Heart	87.51 ± 2.54	2.82 ± 0.31	88.56 ± 2.16	2.78 ± 0.28	89.41 ± 2.45	2.32 ± 0.15
Spleen	90.04 ± 5.63	3.12 ± 0.17	88.75 ± 2.45	2.44 ± 0.27	89.96 ± 3.65	2.75 ± 0.30

Figure 3. Mean (±SD) plasma exposure profile of AG after oral administration at 30 mg/kg in rats.

Figure 4. (a) Concentration of AG in whole blood, plasma and different tissues after multiple oral administration of AG and (b) concentration of AG in whole blood, plasma and different tissues after multiple oral administration of APE.