Bioanalytical methods of hederagenin in rats.

Method	Mobile phase	Gradient program	Internal standard	Linearity	Samples	References
UFLC-MS/MS	Acetate with 5 mM aqueous ammonium (A) and acetonitrile (B)	0–0.3 min, 40–90% B; 0.3–1.9 min, 90% B; 1.9–4.0 min, 90–40% B	Amobar-bital	0.406–203 ng mL−1 (r2 > 0.99)	Plasma; cerebro-spinal fluid	47
HPLC-MS/MS	CH3CN (A) and water containing 0.1% HCOOH (B)	0–3.0 min, 15–40% A; 3.0–7.0 min, 40–65% A; 7.0–9.5 min, 65–88% A; 9.5–13.0 min, 88% A; 13.0–18.0 min, 88–15% A	Warfarin	0.25–500 ng mL−1 (r2 > 0.993)	Plasma	48
HPLC-MS/MS	Acetonitrile (A) and water (B)	Isometric (85 : 15)	Betulonic acid	0.32–64.0 ng mL−1 (r2 > 0.995)	Plasma	49
HPLC-MS/MS	Methanol (A) and 0.05% glacial acetic acid with 10 mM ammonium acetate and 30 μM sodium acetate (B)	0–1.6 min, 73% A; 1.6–1.7 min: 73–89% A; 1.7–5.6 min, 89% A; 5.6–5.8 min, 89–73% A	Glycyrr-hetic acid	2–300 ng mL−1 (r2 > 0.996)	Plasma	50
HPLC-MS/MS	Methanol (A) and 5 mmol L−1 ammonium acetate with 0.025% glacial acetic acid (B)	The first gradient program (applied to brain, kidney, testis, ovary, heart, liver, spleen, lung, fat and muscle): 0–1.9 min, 69% A; 1.9–2.0 min, 69–90% A; 2.0–3.6 min, 90% A; 3.6–3.7 min, 90–69% A; the second gradient program (applied to stomach, large intestine and small intestine): 0–3.7 min, 66% A; 3.7–3.8 min, 66–88.5% A; 3.8–7.8 min, 88.5% A; 7.8–7.9 min, 88.5–100% A; 7.9–11.3 min, 100% A; 11.3–11.4 min, 100–66% A	Glycyrr-hetic acid	2–150 ng mL−1 (r2 > 0.996) for the first gradient program; 3–300 ng mL−1 (r2 > 0.996) for the second gradient program	Tissues including stomach, large intestine, small intestine, brain, kidney, testis, ovary, heart, liver, spleen, lung, fat and muscle	51