Background Ocimum genus, known as Tulsi or Basil, is a prominent botanical class in Asian culture, especially in India. The leaves have immunomodulatory, antioxidant, stress-relieving, and adaptogenic roles in traditional and modern medicine, with prominent usage in herbal teas and nutraceuticals. Objective An high-performance liquid chromatography–photodiode array (HPLC-PDA) method was developed and validated for quantification of vicenin-2, orientin, cynaroside, betulinic acid, genistein with syringic acid, rosmarinic acid, eugenol, carnosic acid, oleanolic acid, ursolic acid, luteolin, and apigenin and was confirmed using a novel electrospray ionisation-mass spectrometry (ESI-MS/MS) method in the Ocimum genus samples. Method The methodology parameters were developed on an reverse phase (RP) C18 column with a gradient elution of 1 mL/min flow rate for 0.1% o-phosphoric acid and acetonitrile at 210 and 340 nm wavelengths. Results The validation data for 13 bioactive compounds showed good linearity (r2 > 0.99) with sensitive LOD (0.034–0.684 µg/mL) and LOQ (0.100–2.068 µg/mL) with recoveries (83.66–101.53%). The results of the quantification were found to be precise (RSD, <5.0%) and accurate (relative error (RE), -0.60–1.06). The method performance was verified by analyzing 10 samples of O. tenuiflorum from the 10 geographical states of India (RSD, <5.0%) and were found to be robust. This HPLC-PDA method with ESI-MS/MS confirmation was applicable to the 13 cultivars from O. thyrsiflorum, O. citriodorum, O. americanum, O. africanum, O. basilicum, O. gratissimum, and O. tenuiflorum species. Conclusions The validated HPLC-PDA and LC-ESI-MS/MS method was found to be selective and suitable for analyzing 13 compounds in O. tenuiflorum and 12 cultivars from the Ocimum genus as a quality control tool. This method can be used in routine analysis as an inexpensive alternative to advanced techniques. Highlights This work is the first to report for vicenin-2, orientin, cynaroside, betulinic acid, and genistein, with simultaneous analysis of eight bioactive compounds in the Ocimum genus.

中文翻译：

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用 LC-ESI-MS/MS 分析罗勒属中的 Vicenin-2、Orientin、Cynaroside、桦木酸、染料木黄酮和主要八种生物活性成分的 RP-HPLC 方法的开发和验证

背景 Ocimum 属，称为 Tulsi 或 Basil，是亚洲文化中的一个重要植物类别，尤其是在印度。叶子在传统和现代医学中具有免疫调节、抗氧化、缓解压力和适应原作用，在凉茶和保健品中具有突出的用途。目的 开发并验证了高效液相色谱-光电二极管阵列 (HPLC-PDA) 方法，用于定量检测vicin-2、荭草苷、紫苏苷、桦木酸、染料木黄酮与丁香酸、迷迭香酸、丁香酚、鼠尾草酸、齐墩果酸、熊果酸、木犀草素和芹菜素，并在罗勒属样品中使用新型电喷雾电离质谱 (ESI-MS/MS) 方法进行了确认。方法 方法学参数是在反相 (RP) C18 色谱柱上开发的，在 210 和 340 nm 波长下以 1 mL/min 的流速梯度洗脱 0.1% 邻磷酸和乙腈。结果 13 种生物活性化合物的验证数据显示出良好的线性 (r2 > 0.99)，具有灵敏的 LOD (0.034–0.684 µg/mL) 和 LOQ (0.100–2.068 µg/mL)，回收率 (83.66–101.53%)。发现定量结果是精确的 (RSD, <5.0%) 和准确的 (相对误差 (RE), -0.60–1.06)。通过分析来自印度 10 个地理州的 10 个 O.tenuiflorum 样品（RSD，<5.0%）验证了该方法的性能，发现该方法的性能是稳健的。该 HPLC-PDA 方法采用 ESI-MS/MS 确认，适用于 13 个栽培品种 O. thyrsiflorum、O. citriodorum、O. americanum、O. Africanum、O. basilicum、O. gratissimum 和 O. tenuiflorum 种。结论 经验证的 HPLC-PDA 和 LC-ESI-MS/MS 方法具有选择性，适合作为质量控制工具分析 O. tenuiflorum 和 12 个罗勒属品种中的 13 种化合物。该方法可用于常规分析，作为先进技术的廉价替代品。亮点 这项工作首次报道了vicin-2、orientin、cynaroside、桦木酸和染料木素，同时分析了罗勒属中的八种生物活性化合物。该方法可用于常规分析，作为先进技术的廉价替代品。亮点 这项工作首次报道了vicin-2、orientin、cynaroside、桦木酸和染料木素，同时分析了罗勒属中的八种生物活性化合物。该方法可用于常规分析，作为先进技术的廉价替代品。亮点 这项工作首次报道了vicin-2、orientin、cynaroside、桦木酸和染料木素，同时分析了罗勒属中的八种生物活性化合物。