Validation of an optimized HPLC/UV method for the quantification of flavonoids in lotus
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ARTICLE
Validation of an optimized HPLC/UV method for the quantification of flavonoids in lotus Ju Sung Lee1, Leo Adrianne Paje1, Won‑Hee Choi2, Eun Ju Cho3, Hyun Young Kim4, Sonia D. Jacinto5 and Sanghyun Lee1*
Abstract Flavonoids present in the leaves of lotus (Nelumbo nucifera) grown in different regions of South Korea (Yeongcheon, Haenam, and Seocheon) and at different harvest times (July to September) were determined. Flavonoid contents in lotus extracts were identified and analyzed using high-performance liquid chromatography (HPLC). The HPLC results revealed that the flavonoid contents of the lotus extracts varied at different harvesting times, with the highest content in July. Analysis of the flavonoid content in the leaves from the different regions showed the highest contents of isorhamnetin-3-O-glucoside, quercetin 3-O-glucuronide, and quercetin 3-O-glucoside in Yeongcheon, Korea, and highest contnts of rutin, myricetin, kaempferol 3-O-glucoside, and quercetin in Haenam, Korea. The HPLC method was validated and optimized to quantify quercetin 3-O-glucuronide; it showed good linearity (1000–62.5 µg/mL, r2 = 0.9999), accuracy (106%–108%), and precision (RSD ≤ 1.70%). Determination of flavonoid content in lotus is valu‑ able for producing medicinal crops and identifying the optimal sources to increase the quantity of clinically available medicines. Keywords: Flavonoid, HPLC, Method, Validation, Nelumbo nucifera Introduction Lotus (Nelumbo nucifera) has a wide distribution throughout North America, Australia, and East Asia [1]. The plant is used for various medicinal purposes and folk remedies in China [2] and is considered to be of cultural and historical importance in Egypt, India, and China [3–5] as it symbolizes perfection, purity, and beauty. In recent years, there have been reports on anti-viral [2], anti-obesity [6], anti-oxidant [7], and lipolytic [8] activities with simultaneous determinations of the components of lotus [9]. As oxygen free radicals, reactive oxygen species (ROS) contribute to aging and many related diseases [10]. The oxygen taken up via breathing is used by mitochondria within the body to generate energy, but they produce byproducts such as ROS which interfere with cell *Correspondence: [email protected] 1 Department of Plant Science and Technology, Chung-Ang University, Anseong 17546, Republic of Korea Full list of author information is available at the end of the article
function; their effects are long-lasting and can eventually lead to cell damage. An excess supply of oxygen produces free radicals that oxidize cells, destroying the cell structure, damaging DNA, and attacking the cell membrane. The resulting damaged cells grow and proliferate and eventually become cancer cells [11]. As ROS are highly unstable biochemical substances, they oxidize fat in the cells and blood to yield lipid peroxides, the main cause of aging. A potential solution to prevent cell damage caused by free radicals is the use of anti-oxidants such as flavonoids. Flavonoids are a large class of
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