Determination of Serum Crocetin After Oral Administration of Krocina Tablets Using HPLC and LC–MS Method

Background: Saffron, derived from Crocus sativus, has long been used in traditional medicine because of its antioxidant, anti-inflammatory, and neuroprotective properties, which are primarily attributed to crocin, its major bioactive pigment. Crocin is extensively hydrolyzed to crocetin in the gastrointestinal tract, where it exhibits improved systemic absorption and acts as the principal circulating metabolite. However, pharmacokinetic data regarding commercially available crocin formulations remain limited. Objectives: Crocin is the main active pigment in saffron and is responsible for its many therapeutic effects. A pharmaceutical product called Krocina is available on the Iran market, and this study aims to investigate the pharmacokinetics and absorption of Krocina ۱۵-mg tablets. Methods: A stock solution of crocetin at a concentration of ۵ µg/mL in methanol was prepared. Serum and urine standards were then prepared from this stock in the concentration range of ۰.۰۲۵ to ۲.۵ µg/mL. Each standard was analyzed with high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) with multiple reaction monitoring (MRM). A mobile phase composed of methanol, water, and acetic acid (۸۵%, ۱۴.۵%, ۰.۵%), at a detection wavelength at ۴۲۳ nm with a flow rate of ۰.۸ mL/min was employed. LC-MS measurements were performed in positive electrospray ionization mode (ESI+) using a Supelco analytical column (۱۵۰×۴.۶ mm, ۳ µm Discovery HS C۱۸, USA). Results: The intraday coefficient of variation (CV) measured by HPLC <۱.۲۸%, and the interday CV<۳.۴۴%. For the LC-MS/MRM method, intraday CV<۱.۰۵% and interday CV was below ۲.۸۷%. The average peak serum concentration of crocetin after Krocina tablet administration occurred at ۲۴۰ minutes, measured as ۱۷۷.۱۱ ng/mL by HPLC and ۱۸۴.۳۳ ng/mL by LC-MS/MRM. Conclusion: Both HPLC and LC-MS methods demonstrated adequate capability for blood analysis and determination of pharmacokinetic parameters, and the obtained data were considered reliable. The clinical effects of Krocina tablets should be related to absorbed crocetin.

کلیدواژه ها:

Krocina ، Tablet ، Pharmacokinetic ، High-performance liquid chromatography (HPLC) ، Liquid chromatography-mass spectrometry (LC-MS)