Liquid chromatography with tandem mass spectrometry is used widely used for the quantitative analysis of phytoconstituents present in medicinal plants to assess the quality of extract used for different investigations. A sensitive, precise, and accurate liquid chromatographic method with tandem mass spectrometric detection was developed for simultaneous quantification of lupeol, betulinic acid, and β-sitosterol in methanolic extract of Madhuca longifolia bark. The three compounds were eluted with stationary phase Gemini C₁₈ column (50mm x 2.0mm, 3μm) and maintained the temperature of the column by column oven at 40 ± 0.3 °C; the composition of mobile phase A (Water and 0.1% formic acid) and mobile phase B (Acetonitrile: Methanol (50:50 v/v) and 0.1% formic acid) in a gradient mode and the flow rate was 0.4 mL/min. With these conditions, the retention time for betulinic acid, lupeol, and β – sitosterol was found to be 1.25, 3.08, and 3.53, respectively. The total run time is 5.0 minutes. Detection and quantitation of all three phytoconstituents were carried out by the mass spectrometer, a triple quadrupole equipped with atmospheric pressure chemical ionization, and multiple reaction monitoring using the predominantly positive ion mode and obtained much higher and more stable response nebulizer gas flow at 3.0 l/min. The linear responses were exhibited for all three phytoconstituents with a dynamic linear range of 10 –100 μg/mL with the values of the regression coefficient more than 0.995 for betulinic acid, lupeol, and β-sitosterol. The values of percentage (%) RSD for intraday and interday precision were found to be within the accepted limits for analytical methods (<15%). Selectivity, linearity, LOD, LOQ, accuracy, and precision were evaluated for all three phytoconstituents as per ICH guidelines. The proposed method is accurate and sensitive and can be used for the routine quantification of betulinic acid, lupeol, and β-sitosterol from the herbal extract and its poly-herbal formulations.

中文翻译：

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的桦木酸同时定量，Lupeolandβ-Sitosterolin长叶通过液相色谱-串联质谱法树皮MethanolicExtract

液相色谱-串联质谱被广泛用于对药用植物中植物成分的定量分析，以评估用于不同研究的提取物的质量。建立了一种灵敏，准确，准确的液相色谱-串联质谱检测方法，用于同时定量长叶马杜鹃树皮甲醇提取物中的羽扇豆酚，桦木酸和β-谷甾醇。用固定相Gemini C ₁₈洗脱这三种化合物色谱柱（50mm x 2.0mm，3μm）并通过柱温箱将色谱柱温度保持在40±0.3°C; 流动相A（水和0.1％的甲酸）和流动相B（乙腈：甲醇（50:50 v / v）和0.1％甲酸）的组成以梯度模式组成，流速为0.4 mL / min。在这些条件下，发现桦木酸，羽扇豆酚和β-谷甾醇的保留时间分别为1.25、3.08和3.53。总运行时间为5.0分钟。通过质谱仪，配备了大气压化学电离的三重四极杆和主要使用正离子模式的多反应监测，对所有三种植物成分进行检测和定量，并在3.0 l / l的条件下获得了更高，更稳定的响应雾化器气流分钟 所有三种植物成分均表现出线性响应，其动态线性范围为10 –100μg/ mL，其中桦木酸，羽扇豆酚和β-谷甾醇的回归系数均大于0.995。发现日内和日间精度的RSD百分比（％）值在分析方法的可接受范围内（<15％）。根据ICH指南评估了所有三种植物成分的选择性，线性，LOD，LOQ，准确性和精密度。所提出的方法准确，灵敏，可用于从草药提取物及其多草药配方中对桦木酸，羽扇豆醇和β-谷甾醇进行常规定量分析。发现日内和日间精度的RSD百分比（％）值在分析方法的可接受范围内（<15％）。根据ICH指南评估了所有三种植物成分的选择性，线性，LOD，LOQ，准确性和精密度。所提出的方法准确，灵敏，可用于从草药提取物及其多草药配方中对桦木酸，羽扇豆醇和β-谷甾醇进行常规定量分析。发现日内和日间精度的RSD百分比（％）值在分析方法的可接受范围内（<15％）。根据ICH指南评估了所有三种植物成分的选择性，线性，LOD，LOQ，准确性和精密度。所提出的方法准确，灵敏，可用于从草药提取物及其多草药配方中对桦木酸，羽扇豆醇和β-谷甾醇进行常规定量分析。