Journal of Analytical Methods in Chemistry

Research Article

Metabolite Profiling, Pharmacokinetics, and In Vitro Glucuronidation of Icaritin in Rats by Ultra-Performance Liquid Chromatography Coupled with Mass Spectrometry

Table 1

UPLC-MS analysis of icaritin and its observed metabolites in biosamples.


Number		UV	[M + H]⁺	Error	Formula	(+) ESI-MS/MS	(–) ESI-MS/MS	Sources	Characterization
Number	min	nm	[M + H]⁺	ppm	Formula	(+) ESI-MS/MS	(–) ESI-MS/MS	Sources	Characterization

M0^#	7.20	272	369.1336	–0.5	C₂₁H₂₀O₆	391.1152 369.1336 313.0714 215.0748	367.1180 352.0941 309.0417 297.0399 281.0443 253.0485	PUBFI	Icaritin
M1	2.48	270	721.1978	–0.3	C₃₃H₃₆O₁₈	743.1882 721.1978 545.1666 369.1345 313.0717	719.1825 543.1521 367.1194	PUBI	Icaritin-di-O-gluA
M2	2.62	269	563.1768	0.5	C₂₇H₃₀O₁₃	563.1768 387.1442 369.1332 313.0716	561.1608 385.1288	UBI	Hydrated icaritin-O-gluA
M3	2.89	345	531.1511	1.5	C₂₆H₂₆O₁₂	531.1511 355.1166 299.0576	529.1348 353.1027 119.0504	UBI	Desmethylicaritin-O-gluA
M4	3.15	341	561.1617	1.6	C₂₇H₂₈O₁₃	561.1617 385.1301 367.1186	559.1456 383.1175 367.1135 175.0263	UBI	Hydroxylated icaritin-O-gluA
M5	3.22	341	561.1613	0.9	C₂₇H₂₈O₁₃	561.1613 385.1296	559.1458 383.1172 175.0262	UBI	Hydroxylated icaritin-O-gluA
M6	3.36	345	531.1503	0.0	C₂₆H₂₆O₁₂	531.1503 355.1175 329.0741 314.0455	529.1345 353.1025 119.0506	UBI	Desmethylicaritin-O-gluA
M7	3.41	341	561.1605	–0.5	C₂₇H₂₈O₁₃	561.1605 385.1295 313.0715	559.1460 383.1171	BI	Hydroxylated icaritin-O-gluA
M8	3.46	270	721.1982	0.3	C₃₃H₃₆O₁₈	721.1982 545.1662 369.1340 313.0717	719.1823 543.1519 367.1196	UBI	Icaritin-di-O-gluA
M9	3.52	341	561.1608	0	C₂₇H₂₈O₁₃	561.1608 385.1283	559.1454 383.1170 175.0260	BI	Hydroxylated icaritin-O-gluA
M10	3.58	269	563.1761	–0.7	C₂₇H₃₀O₁₃	563.1761 387.1452 369.1350 313.0735	561.1605 385.1289	UBI	Hydrated icaritin-O-gluA
M11	3.74	341	561.1615	1.2	C₂₇H₂₈O₁₃	561.1615 385.1292 367.1186	559.1462 383.1175	BI	Hydroxylated icaritin-O-gluA
M12	4.29	n.a.	543.1501	–0.4	C₂₇H₂₆O₁₂	543.1501 367.1189	541.1343 365.1034 351.0765 175.0221 113.0253	I	Dehydrogenated icaritin-O-gluA
M13	4.34	271	545.1663	0.7	C₂₇H₂₈O₁₂	567.1486 545.1663 369.1335 313.0716	543.1501 367.1188 352.0948 309.0408 297.0398 281.0498	PUBFI	Icaritin-O-gluA
M14^#	4.40	270	823.3024	–0.1	C₃₉H₅₀O₁₉	823.3024 369.1324 313.0721	821.2863 659.2331 367.1168 351.0867	I	Epimedin C
M15	4.45	273	387.1440	–1.0	C₂₁H₂₂O₇	387.1440 369.1338 313.0722	385.1288	I	Hydrated icaritin
M16	4.49	270	823.3021	–0.5	C₃₉H₅₀O₁₉	823.3021 369.1336 313.0725	821.2860 659.2333 367.1165 351.0869	I	Epimedin C isomer
M17^#	4.53	270	531.1866	1.5	C₂₇H₃₀O₁₁	531.1866 369.1350 313.0718	529.1705 367.1182 352.0948 297.0395 253.0496	FI	Icariside I
M18	4.57	271	545.1661	0.4	C₂₇H₂₈O₁₂	545.1661 369.1346 313.0723	543.1505 367.1183 352.0945 297.0394	PUBFI	Icaritin-O-gluA
M19	4.59	271	647.2349	1.4	C₃₂H₃₈O₁₄	669.2215 647.2349 515.1930 369.1346 313.0724	645.2168 367.1162 351.0863 323.0945 295.0595	I	Icaritin-rha-xyl
M20	4.61	270	661.2503	1.1	C₃₃H₄₀O₁₄	683.2326 661.2503 369.1339 313.0719	659.2343 367.1165 351.0865 295.0602 217.0499	FI	Icaritin-rha-rha
M21^#	4.88	269	515.1927	1.9	C₂₇H₃₀O₁₀	537.1757 515.1927 369.1340 313.0726	513.1766 367.1157 352.0900 323.0910 295.0604 217.0498	FI	Icariside II
M22	4.93	300	543.1500	–0.6	C₂₇H₂₆O₁₂	565.1453 543.1500 367.1172	541.1345 365.1033 175.0222 113.0254	BI	Dehydrogenated icaritin-O-gluA
M23	4.97	273	387.1445	0.3	C₂₁H₂₂O₇	387.1445 369.1343 313.0715	385.1286	UFI	Hydrated icaritin
M24	5.02	300	543.1505	0.4	C₂₇H₂₆O₁₂	543.1505 367.1134 313.0720	541.1346 365.1032 175.0223 113.0255	BI	Dehydrogenated icaritin-O-gluA
M25^#	5.08	270	355.1180	–0.6	C₂₀H₁₈O₆	355.1180 299.0582	353.1036 309.0409 297.0373 281.0481	UFI	Desmethylicaritin
M26	5.13	271	545.1664	0.9	C₂₇H₂₈O₁₂	567.1540 545.1664 369.1343 313.0720 299.0585	543.1504 367.1180 352.0942 297.0391	BI	Icaritin-O-gluA
M27	5.37	268	385.1288	0.3	C₂₁H₂₀O₇	407.1136 385.1288 367.1186 313.0715	383.1175 365.1035 175.0226	UFI	Hydroxylated icaritin
M28	5.54	268	385.1285	–0.5	C₂₁H₂₀O₇	385.1285 367.1181 313.0719	383.1170 365.1046	UFI	Hydroxylated icaritin
M29	5.66	n.a.	385.1290	0.9	C₂₁H₂₀O₇	385.1290 367.1176 313.0712	383.1172 367.1043 175.0262	UFI	Hydroxylated icaritin
M30	6.36	272	367.1180	–0.5	C₂₁H₁₈O₆	367.1180 352.0948 313.0721 297.0395 253.0496	365.1047 351.0765 175.0223 113.0250	FI	Dehydrogenated icaritin

M0, parent drug; M1~M34, metabolites; n.a., not available; P, plasma; U, urine; B, bile; F, feces; I, intestine. gluA, glucuronide conjugates; glc, glucose; rha, rhamnose; xyl, xylose.
# means that the metabolites are exactly identified with reference standards.