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CAS:64-77-7
Appearance:White to off-white powder
Storage:2-8℃
Purity:HPLC≥98%
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LC-MS/MS
The chromatographic separation was carried out using an C18, 2.1×100 mm , 3.5-μm column, on a LC-30A UFLC system. The column oven temperature was set at 35 C. A mobile phase consisting of 0.025 % formic acid in water (A) and methanol (B) was delivered at a flow rate of 0.2 mL/min using the gradient program. The mass operation parameters were: ion source gas 1 was set at 55 psi, ion source gas 2 at 60 psi, curtain gas at 30 psi, and collision gas at medium.The gradient program using was: 15 % (B) from 0 to 2 min,15-55 % (B) from 2 to 6 min, 55-75 % (B) from 6 to 16 min, 75 % (B) from 16 to 17 min, 75-15 % (B) from 17 to 17.5 min and 15 % (B) from 17.5 to 22 min. A negative Turbo Spray ion source was used in the MS analysis. Temperature was set at 600 C and IonSpray voltage at -4500V. In product ion scan mode, product ions of m/z 663.4, 839.4, 825.4 were searched respectively. Scan range was set at 100-700 Da for m/z 663.4, and 100-850 Da for m/z 839.4 and 825.4. In precursor ion scan mode, precursor ions of m/z 663.4 and 501.3 were searched respectively. Scan range was set at 300-1100, 670-1100 and 750-1100 Da. In both product ion scan mode and precursor ion scan mode, scan rate was set at 200 Da/s, declustering potential (DP) start and DP stop at -100 and -150V, and collision energy (CE) at -36V. In multiple reaction monitoring (MRM) mode, ilexsaponin A1 and its two potential metabolites were monitored simultaneously. The precursor-to-product ion pairs for MRM transitions, DP and CE were optimized at m/z 663.4→ 501.3, -150V and -45V for ilexsaponin A1, m/z 839.4 → 501.3, -40 V and -70 for M1 (a glucuronic acid conjugation product), and m/z 825.4→ 501.3, -70 V and -60V for M2 (a glucose conjugation product), respectively. MRM mode was also used in both metabolic stability study and metabolic enzyme identification, including the precursor-to-product ion pairs for MRM transitions, DP and CE set for pedunculoside (IS) were at m/z 695.4 → 487.3, -120V and -36 V, respectively.
References：
Wu L, Kang A, Jin X, Bao Y, Miao P, Lv T, Zhou Z. Ilexsaponin A1: In vitro metabolites identification and evaluation of inhibitory drug-drug interactions. Drug Metab Pharmacokinet. 2021 Oct;40:100415. doi: 10.1016/j.dmpk.2021.100415. Epub 2021 Aug 2. PMID: 34461570.
