Development of UPLC-MS/MS Method for Studying the Pharmacokinetic Interaction Between Dasatinib and Posaconazole in Rats

Figures & data

Figure 1 Mass spectra and structures of dasatinib (A) and posaconazole (B) in present study.

Table 1 Gradient Elution Programme of HPLC for Dasatinib and Posaconazole

Time	0.1% Aqueous Formic Acid (Composition, %)	Acetonitrile (Composition, %)	Flow Rate
0	90	10	0.3 mL/min
0.5	90	10
1.0	10	90
2.0	10	90
2.5	90	10
3.0	90	10

Figure 2 Representative liquid chromatograms of dasatinib, posaconazole, and IS in rat plasma sample. (A) blank plasma; (B) plasma standard of 10 ng/mL dasatinib and 50 ng/mL posaconazole; (C) plasma sample from combined treatment group 5 h after orally administration of dasatinib and posaconazole simultaneously.

Table 2 Accuracy, Precision, Matrix Effect and Recovery for the Analysis of Dasatinib and Posaconazole in Rat Plasma (n=6)

Analyte	Concentration (ng/mL)	Accuracy (RE, %)		Precision (RSD, %)		Matrix Effect (%)	Recovery (%)
		Intra-Day	Inter-Day	Intra-Day	Inter-Day
Dasatinib	1	3.17	2.97	7.99	9.17	-	-
	3	5.84	−2.53	7.65	7.45	88.71±2.54	89.87±4.32
	500	9.08	6.86	2.69	3.54	94.19±6.9	87.85±2.32
	750	5.23	7.06	5.36	3.85	92.47±7.24	88.93±3.27
Posaconazole	5	7.29	4.21	6.74	10.11	-	-
	15	6.36	7.39	7.44	6.93	91.92±8.41	86.20±8.02
	2500	6.31	8.12	6.48	3.53	100.96±5.86	78.50±9.09
	3750	2.51	4.34	8.53	3.88	98.57±7.74	77.06±8.40

Table 3 The stability for the Analysis of Dasatinib and Posaconazole in Rat Plasma (n=6)

Analytes	Concentration (ng/mL)	Stability (%)
		Ambient, 12 h	Autosampler, 6 h	Three Freeze-Thaw Cycles	−80 °C, 30 Days
Dasatinib	3	7.42	8.42	5.35	5.12
	750	6.03	3.92	4.68	5.88
Posaconazole	15	3.96	5.6	7.76	6.21
	3750	5.82	4.2	4.14	2.95

Table 4 The Main Pharmacokinetic Parameters of Dasatinib and Posaconazole in Three Groups (n = 8 Each Group)

Parameters	Group A	Group B	Group C
			Dasatinib	Posaconazole
AUC0-t (ng*h/mL)	1350.86 (878.02–2376.38)	7528.97 (4217.76–17,907.69)	1395.6 (1059.6–1905.2)	73,856.23 (25,260.08–97,420.83)**
AUC0-∞ (ng*h/mL)	1371.08 (878.2–2376.44)	7617.16 (4533.24–18,087.2)	1677.42 (1155.2–2185.62)	78,809.65 (26,028.33–121,006.9)**
t1/2z (h)	4.52 (3.04–14)	12.43 (9.63–23.75)	9.63 (3.06–19.92)	15.25 (10.72–24.33)
Tmax (h)	1.75 (1–5)	8 (4–12)	1.375 (1–3)	24 (12–24)**
CLz/F(L/h/kg)	7.36 (4.21–11.39)	5.46 (2.21–8.82)	5.97 (4.58–8.66)	0.51 (0.33–1.54)**
Cmax (ng/mL)	189.72 (108.48–399.19)	390.27 (204.63–892.45)	175.91 (131.62–403.32)	2329.47 (1119.42–2680.17)**

Note: **p < 0.01, significant in comparison with group A or group B.

Abbreviations: AUC_0-t, area under the curve from time zero to the last quantifiable concentration; AUC_0-∞, area under the curve from time zero to infinity; t_1/2z, half-life; T_max, time to maximum concentration; CL_z/F, clearance; C_max, maximal plasma concentration.

Figure 3 Mean concentration-time curves of dasatinib and posaconazole in dasatinib treatment alone group, posaconazole treatment alone group, and two drugs combined treatment group (A and B). Group A: 10 mg/kg dasatinib; group B: 40 mg/kg posaconazole; group C: 10 mg/kg dasatinib plus 40 mg/kg posaconazole. Each time-point represents the mean±SD.