Targeted Ultra‐High Performance Liquid Chromatography–Electrospray Ionization Triple Quadrupole Mass Spectrometry‐Based Quantification of Diosgenin in <i>Hellenia speciosa</i> for Its Wild Populations Screening: A Quality Control Assessment

ABSTRACT Hellenia speciosa rhizome is a natural source of diosgenin for steroidal drug synthesis. However, there is very scanty information available on the quantification of natural diosgenin using targeted ultra‐high performance liquid chromatography–electrospray ionization triple quadrupole–mass spectrometry (UHPLC–ESI–QqQ–MS/MS) method. Therefore, the present study aimed to develop and validate the targeted UHPLC–ESI–QqQ–MS/MS method to quantify diosgenin for the quality control of the wild H. speciosa populations of Telangana. The developed UHPLC–ESI–QqQ–MS/MS in positive mode showed good linearity (0.02–2.0 µg/mL) for diosgenin in triplicate analysis with good correlation coefficient ( y = 1549.4 x − 41.982, R 2 = 0.998). The limit of detection (LOD) and limit of quantification (LOQ) for diosgenin were found to be 0.06 and 0.18 µg/mL, respectively. The method was accurate and mean recovery percentage for diosgenin ranged from 101.17% to 101.99% with %RSD from 0.78 to 1.86. The intra‐day and inter‐day variations were found to be 0.95% and 1.12%, which confirmed high precision with repeatability and reproducibility. The developed method was robust as there is no impact of change in pH of mobile phase and column oven temperature. The quality assessment of wild H. speciosa rhizomes 14 populations showed significant ( p &lt; 0.05) variation in the total ash (7.7%–19.9%), acid insoluble ash (0.3%–5%), alcohol soluble extractive (4.1%–8.6%), water‐soluble extractive (0.9%–6.9%), total alkaloids (0.85%–4.7%), total flavonoids (3.0%–7.6%), total terpenoid (1.3%–5.3%), total saponin (2.4%–8.2%), methanol extract (8.6%–15.3%), and diosgenin (0.008%–0.792%). A Pearson correlation revealed significant correlation of total saponin ( p &lt; 0.01) with extract yield (EY) ( r = 0.62) and diosgenin content ( r = 0.77). Principal component analysis confirmed H. speciosa 06 as a promising population with higher saponin, terpenoid, EY, and diosgenin contents. Overall, the developed UHPLC–ESI–QqQ–MS/MS provides rapid and easy method for the quality control of H. speciosa .