Department of Pharmaceutical Sciences, College of Clinical Pharmacy, King Faisal University, Al-Ahsa 31982, Saudi Arabia.
Department of Pharmacognosy, Institute of Pharmacy, Nirma University, Ahmedabad 382481, Gujarat, India.
Molecules. 2020 Oct 26;25(21):4947. doi: 10.3390/molecules25214947.
Sinigrin, a precursor of allyl isothiocyanate, present in the exhibits diverse biological activities, and has an immense role against cancer proliferation. Therefore, the objective of this study was to quantify the sinigrin in the roots using developed and validated RP-HPLC method and further evaluated its' anticancer activity. To achieve the objective, the roots of were lyophilized to obtain a stable powder, which were extracted and passed through an ion-exchange column to obtain sinigrin-rich fraction. The RP-HPLC method using C18 analytical column was used for chromatographic separation and quantification of sinigrin in the prepared fraction, which was attained using the mobile phase consisting of 20 mM tetrabutylammonium: acetonitrile (80:20%, / at pH 7.0) at a flow rate of 0.5 mL/min. The chromatographic peak for sinigrin was showed at 3.592 min for pure sinigrin, where a good linearity was achieved within the concentration range of 50 to 800 µg/mL ( > 0.99), with an excellent accuracy (-1.37% and -1.29%) and precision (1.43% and 0.94%), for intra and inter-day, respectively. Finally, the MTT assay was performed for the sinigrin-rich fraction using three different human cancer cell lines, viz. prostate cancer (DU-145), colon adenocarcinoma (HCT-15), and melanoma (A-375). The cell-based assays were extended to conduct apoptotic and caspase-3 activities, to determine the mechanism of action of sinigrin in the treatment of cancer. MTT assay showed values of 15.88, 21.42, and 24.58 µg/mL for DU-145, HCT-15, and A-375 cell lines, respectively. Increased cellular apoptosis and caspase-3 expression were observed with sinigrin-rich fraction, indicating significant increase in overexpression of caspase-3 in DU-145 cells. In conclusion, a simple, sensitive, fast, and accurate RP-HPLC method was developed for the estimation of sinigrin in the prepared fraction. The data observed here indicate that sinigrin can be beneficial in treating prostate cancer possibly by inducing apoptosis.
白芥子苷，异硫氰酸烯丙酯的前体，存在于 中，具有多种生物活性，对癌症增殖具有重要作用。因此，本研究的目的是使用已建立和验证的反相高效液相色谱法（RP-HPLC）定量 根中的白芥子苷，并进一步评估其抗癌活性。为了实现这一目标，将 的根冻干以获得稳定的粉末，然后用离子交换柱提取并通过该柱以获得富含白芥子苷的部分。使用 C18 分析柱的 RP-HPLC 方法用于分离和定量制备部分中的白芥子苷，该方法使用包含 20 mM 四丁基铵：乙腈（80:20%，/在 pH 7.0 下）的流动相以 0.5 mL/min 的流速进行。白芥子苷的色谱峰在纯白芥子苷的 3.592 分钟处显示，在 50 至 800 µg/mL 的浓度范围内（> 0.99）具有良好的线性关系，日内和日间精度分别为-1.37%和-1.29%和 1.43%和 0.94%。最后，使用三种不同的人癌细胞系（前列腺癌（DU-145）、结肠腺癌（HCT-15）和黑色素瘤（A-375））对富含白芥子苷的部分进行 MTT 测定。将细胞测定扩展到进行细胞凋亡和 caspase-3 活性，以确定白芥子苷在癌症治疗中的作用机制。MTT 测定法显示 DU-145、HCT-15 和 A-375 细胞系的 值分别为 15.88、21.42 和 24.58 µg/mL。用富含白芥子苷的部分观察到细胞凋亡增加和 caspase-3 表达增加，表明 DU-145 细胞中 caspase-3 的过表达显著增加。总之，开发了一种简单、灵敏、快速和准确的 RP-HPLC 方法来估计制备部分中的白芥子苷。这里观察到的数据表明，白芥子苷通过诱导细胞凋亡可能对治疗前列腺癌有益。
