School of Pharmaceutical Sciences, Lovely Professional University, Jalandhar-Delhi G.T Road, Phagwara 144411, Punjab, India.
Department of Pharmacology, Umm Al-Qura College of Pharmacy, Umm Al-Qura University, Makkah 21955, Saudi Arabia.
Molecules. 2022 Oct 21;27(20):7117. doi: 10.3390/molecules27207117.
Xanthohumol (XH) a prenylated chalcone has diverse therapeutic effects against various diseases. In the present study, a bioanalytical method was developed for XH in rat plasma using reverse phase high performance liquid chromatography. The validation of the method was performed as per ICH M10 guidelines using curcumin as an internal standard. The Isocratic elution method was used with a run time of 10 min, wherein the mobile phase ratio 0.1% / OPA (A): Methanol (B) was 15:85 / at flow rate 0.8 mL/min and injection volume of 20 µL. The chromatograms of XH and curcumin was recorded at a wavelength of 370 nm. The retention time for XH and curcumin was 7.4 and 5.8 min, respectively. The spiked XH from plasma was extracted by the protein precipitation method. The developed method was linear with R value of 0.9996 over a concentration range of 50-250 ng/mL along with LLOQ. The results of all the validation parameters are found to be within the accepted limits with %RSD value less than 2 and the percentage recovery was found to be greater than 95%. Based on the %RSD and percentage recovery results it was confirmed that the method was precise and accurate among the study replicates. LOD and LOQ values in plasma samples were found to be 8.49 ng/mL and 25.73 ng/mL, respectively. The stability studies like freeze thaw, short term and long-term stability studies were also performed, %RSD and percentage recovery of the XH from plasma samples were within the acceptable limits. Therefore, the developed bioanalytical method can be used effectively for estimation of XH in plasma samples.
黄腐酚（XH）是一种具有不同治疗作用的类查耳酮，可用于多种疾病。在本研究中，建立了大鼠血浆中黄腐酚的反相高效液相色谱分析方法。该方法的验证符合 ICH M10 指南，以姜黄素为内标。采用等度洗脱法，运行时间为 10 分钟，其中流动相比例为 0.1%/OPA（A）：甲醇（B）为 15:85/流速为 0.8 mL/min，进样量为 20 µL。XH 和姜黄素的色谱图在 370nm 处记录。XH 和姜黄素的保留时间分别为 7.4 和 5.8 分钟。血浆中的 XH 经蛋白沉淀法提取。该方法在 50-250ng/mL 的浓度范围内具有线性关系，R 值为 0.9996，LLOQ 也具有线性关系。所有验证参数的结果均在可接受范围内，%RSD 值小于 2，回收率大于 95%。基于%RSD 和回收率结果，证实该方法在研究重复中具有较高的精密度和准确度。血浆样品中的 LOD 和 LOQ 值分别为 8.49ng/mL 和 25.73ng/mL。还进行了冻融、短期和长期稳定性研究等稳定性研究，血浆样品中 XH 的%RSD 和回收率均在可接受范围内。因此，所建立的生物分析方法可有效地用于血浆样品中黄腐酚的定量分析。
