RP-HPLC-PDA method has been developed and validated for the quantification of Steviol glycoside: Rebaudioside A in aqueous extract of Stevia rebaudiana plant leaf. Rebaudioside A was separated on a Phenomenex Luna C18 analytical column with column temperature 40 °C using an isocratic solvent system consisting (70:30 v/v) mixture of 10 mmol/L sodium phosphate buffer and acetonitrile having pH-2.6 at 1.0 mL/min flow rate with detection at 210 nm wavelength. The method has been validated for linearity, precision, accuracy, detection limit, quantitation limit and robustness. The proposed method linearity for Rebaudioside A with a regression coefficient of 0.9999 was obtained within the range of 0.0005–1.5 mg/mL. The relative standard deviation was less than 2.0% in intra-day and inter-day precision. The proposed method’s accuracy was determined by a three different level recovery study. The recovery average was between 95.20 and 96.77 percent. Rebaudioside A levels of LOD and LOQ were 0.01 and 0.05 mg/mL respectively. The leaves from eight different Indian territories were extracted by sonication extraction method. Rebaudioside A content obtained in the dried leaves was within the range of 2.39–7.53 percent w/w. The method proposed is simple, rapid and reproducible; therefore, it is appropriate for routine analysis of Rebaudioside A in plant material as well as finished products which are used commercially as high-intensity sweeteners across the globe to replace sugar.

中文翻译：

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一种用于定量甜菊糖苷的经过验证的 RP-HPLC 方法：甜菊叶提取物中的莱鲍迪苷 A

已开发并验证了 RP-HPLC-PDA 方法，用于定量甜菊糖苷：甜叶菊植物叶水提取物中的莱鲍迪苷 A。在 Phenomenex Luna C18 分析柱上分离莱鲍迪苷 A，柱温为 40 °C，使用等度溶剂系统（70:30 v/v）由 10 mmol/L 磷酸钠缓冲液和 pH-2.6 的乙腈混合物组成，1.0 mL/在 210 nm 波长下检测时的最小流速。该方法已经过线性、精密度、准确度、检测限、定量限和稳健性的验证。在 0.0005–1.5 mg/mL 范围内获得了回归系数为 0.9999 的莱鲍迪苷 A 的建议方法线性。日内和日间精密度的相对标准偏差小于2.0%。建议方法的准确性由三个不同级别的回收研究确定。平均回收率在 95.20% 和 96.77% 之间。莱鲍迪苷 A 的 LOD 和 LOQ 水平分别为 0.01 和 0.05 mg/mL。通过超声提取法提取了来自八个不同印度领土的叶子。在干燥的叶子中获得的莱鲍迪苷 A 含量在 2.39-7.53% w/w 的范围内。所提出的方法简单、快速、重现性好；因此，它适用于植物材料和成品中莱鲍迪苷 A 的常规分析，这些产品在全球范围内用作高强度甜味剂以替代糖。通过超声提取法提取了来自八个不同印度领土的叶子。在干燥的叶子中获得的莱鲍迪苷 A 含量在 2.39-7.53% w/w 的范围内。所提出的方法简单、快速、重现性好；因此，它适用于植物材料和成品中莱鲍迪苷 A 的常规分析，这些产品在全球范围内用作高强度甜味剂以替代糖。通过超声提取法提取了来自八个不同印度领土的叶子。在干燥的叶子中获得的莱鲍迪苷 A 含量在 2.39-7.53% w/w 的范围内。所提出的方法简单、快速、重现性好；因此，它适用于植物材料和成品中莱鲍迪苷 A 的常规分析，这些产品在全球范围内用作高强度甜味剂以替代糖。