Spinacia oleracea (S. oleracea), also known as spinach or Indian spinach, is a rich source of carotenoids
and flavonoids with a wide range of health benefits. Besides S. oleracea, many other plants are also known as spinach
or its common versions, which could be misleading in the identification
of S. oleracea in any nutraceuticals and dietary
supplements. Thus, the present study aims to differentiate S. oleracea from other conventionally known spinach plants
by its macroscopic, microscopic, and phytochemical features. Although
these plants were morphologically distinct, analytical investigations
were useful in differentiating S. oleracea from other
spinach plants in dried or powdered forms. In the present study, HPTLC
fingerprinting could efficiently differentiate between these plants
based on the isoflavones present in the leaves of S. oleracea. Followed by a robust and simple HPLC-PDA method that was validated
for 20β- hydroxyecdysone and isoflavones as authenticating markers
with kaempferol from S. oleracea leaves for quality
control. These selected bioactive markers specific to S. oleracea were present in the range of 3.101–1232.820 mg/kg and were
found absent in spinach samples. Essentially, the described set of
methods could be used for species authentication and standardization
of S. oleracea differing from other plants known
as spinach.