Objective: To develop a supercritical fluid chromatography method for the simultaneous determination of active pharmaceutical ingredient (API)-calcitriol, as well as two antioxidants-butylated hydroxy toluene (BHT) and butylated hydroxyanisole (BHA), in calcitriol soft capsules. Methods: Calcitriol, BHT and BHA were separated and determined on an ACQUITY UPC2 Torus DIOL column (100 mm×3.0 mm,1.7 μm) maintained at 40 ℃ with mobile phase containing a mixture of CO2 and methanol at 1.0 mL·min-1 in gradient mode. The detection wavelength was set at 265 nm, the back pressure was set at 13.8 MPa and injection volume was 2 μL. Results: BHT, BHA and calcitriol were successfully separated within 5 min with satisfied resolutions. Good linear relationships were established by plotting the peak area versus concentration in the range of 17.0-85.1 μg·mL-1 for BHT, 17.1-85.3 μg·mL-1 for BHA and 0.34-1.71 μg·mL-1 for calcitriol, respectively (r=0.999 9, 0.999 7 and 0.999 6, respectively, n=5). The quantitation limits (LOQs) were 3.5 ng for BHT, 2.5 ng for BHA and 0.2 ng for calcitriol (S/N=10), respectively. The average recoveries (n=9) of BHT, BHA and calcitriol were 101.5%, 101.8% and 98.3% with RSDs of 1.8%, 2.1% and 1.2%, respectively. The average contents were 0.266, 0.264 and 0.272 μg per capsule for calcitriol,15.6, 15.6 and 15.8 μg per capsule for BHT, and 17.0, 16.9 and 17.0 μg per capsule for BHA in three batches of calcitriol soft capsules. Conclusion: The proposed environmentally friendly method demonstrates high efficiency, accuracy, repeatability, and stability. It can be employed for the quality control and stability research of calcitriol, BHT and BHA in calcitriol soft capsules, which might provide a reference for the quality control for other lipophilic drugs.
ZHAN Bing, JIN Wei, LI Dan, YANG Yong-jian, LE Jian
. Simultaneous determination of calcitriol and antioxidants in calcitriol soft capsules[J]. Chinese Journal of Pharmaceutical Analysis, 2023
, 43(7)
: 1255
-1262
.
DOI: 10.16155/j.0254-1793.2023.07.20
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