Natural Product Chemistry and Process Development Division and AcSIR, CSIR-Institute of Himalayan Bioresource Technology, Palampur, Himachal Pradesh, India.
Phytochem Anal. 2020 Nov;31(6):861-873. doi: 10.1002/pca.2951. Epub 2020 Jun 3.
Trillium govanianum (Nag Chhatri and Teen Patra) is traditionally used for curing joint pains, wounds, and sexual disorders. Steroidal saponins are the main active components of this species. However, only a small amount of information is available about steroidal saponins of this plant.
To develop an ultra-high-performance liquid chromatography-quadrupole time of flight tandem mass spectrometry (UHPLC-QTOF-MS/MS) and ultra-high-performance liquid chromatography-evaporative light scattering detector (UHPLC-ELSD) methods for the qualitative and quantitative determination of steroidal saponins in T. govanianum.
The dried rhizomes of T. govanianum (100 mg) were extracted with ethanol-water (80:20, 10 mL) by ultrasonic treatment for 30 min at 40°C. The prepared sample was analysed by UHPLC-QTOF-MS/MS and UHPLC-ELSD for the qualitative and quantitative determination of steroidal saponins.
A total of 24 saponins were identified using UHPLC-QTOF-MS/MS; seven of them were characterised by comparing with standards. Furthermore, five saponins [govanoside B (2), protodioscin (6), pennogenin tetraglycosides (11), borassoside E (21) and borassoside D (24)] were quantified using UHPLC-ELSD method in different extracts and fractions of T. govanianum. The method showed good linearity (R ≥ 0.993), limit of detection (0.92-4.09 μg/mL), limit of quantification (3.1-13.5 μg/mL), precision [intra-day relative standard deviations (RSDs) < 4.3% and inter-day RSDs < 5.5%], and accuracy (84.0-110.3%). This is the first report on the quantification of 2, 6, 11, 21 and 24 in T. govanianum.
The present study provides an efficient analytical method for the identification and quantification of steroidal saponins and will be helpful for the quality evaluation of T. govanianum.
延龄草（Nag Chhatri 和 Teen Patra）传统上用于治疗关节疼痛、伤口和性障碍。甾体皂苷是该物种的主要活性成分。然而，关于这种植物的甾体皂苷，只有少量信息。
建立一种超高效液相色谱-四极杆飞行时间串联质谱（UHPLC-QTOF-MS/MS）和超高效液相色谱-蒸发光散射检测器（UHPLC-ELSD）方法，用于定性和定量测定延龄草中的甾体皂苷。
将延龄草根茎（100 mg）用 80:20（v/v）乙醇-水（10 mL）在 40°C 下超声处理 30 min 提取。用 UHPLC-QTOF-MS/MS 和 UHPLC-ELSD 分析制备的样品，以定性和定量测定甾体皂苷。
利用 UHPLC-QTOF-MS/MS 共鉴定出 24 种皂苷；其中 7 种通过与标准品比较进行了鉴定。此外，还利用 UHPLC-ELSD 法对延龄草不同提取物和馏分中的 5 种皂苷[govanoside B（2）、薯蓣皂苷元（6）、原薯蓣皂苷（11）、波拉萨苷 E（21）和波拉萨苷 D（24）]进行了定量。该方法具有良好的线性（R≥0.993）、检测限（0.92-4.09μg/mL）、定量限（3.1-13.5μg/mL）、精密度[日内相对标准偏差（RSD）<4.3%和日间 RSD<5.5%]和准确度（84.0-110.3%）。这是首次报道延龄草中 2、6、11、21 和 24 的定量分析。
本研究为鉴定和定量测定甾体皂苷提供了一种高效的分析方法，将有助于延龄草的质量评价。
