Optimization of Ultrasound-Assisted Extraction Conditions Followed by Solid Phase Extraction Fractionation from Orthosiphon stamineus Benth (Lamiace) Leaves for Antiproliferative Effect on Prostate Cancer Cells.

Siti Hasyimah Suhaimi,Hassan Fahmi Ismail,Mohamad Khairul Hafiz Idris,Rosnani Hasham,Fadzilah Adibah Abdul Majid,Nor Hazwani Mohd Ariffin

doi:10.3390/molecules24224183

Abstract

Primarily, optimization of ultrasonic-assisted extraction (UAE) conditions of Orthospihon stamineus was evaluated and verified using a central composite design (CCD) based on three factors including extraction time (minutes), ultrasound amplitude (A), and solvent concentration (%). The response surface methodology (RSM) was performed to develop an extraction method with maximum yield and high rosmarinic acid content. The optimal UAE conditions were as follows: extraction time 21 min, ultrasound amplitudes 62 A, and solvent composition 70% ethanol in water. The crude extract was further fractionated using solid-phase extraction (SPE), where six sequential fractions that varied in polarity (0–100% Acetonitrile in water) were obtained. Next, the six fractions were evaluated for their antioxidant and anti-cancer properties. This study found that Fraction 2 (F2) contained the highest rosmarinic acid content and showed the strongest antioxidant activity. Additionally, F2 showed an anti-proliferative effect against prostate cancer (DU145) with no harmful effect on normal cells.

Highlights

Prostate cancer is a lethal disease occuring in men
Extraction time is a crucial parameter in solvent extraction as it can reduce time and save cost
The yield of rosmarinic acid increased with the increased extraction time from 5 min to 20 min

Summary

Introduction

Prostate cancer is a lethal disease occuring in men. The development of this disease is influenced by various factors such as androgen level, obesity, chronic inflammation, genetics, age, and ethnicity [1].Clinical treatments of cancer are available, which include hormonal therapy, chemotherapy, radiation therapy, and chemotherapie drugs [2]. Prostate cancer is a lethal disease occuring in men. The development of this disease is influenced by various factors such as androgen level, obesity, chronic inflammation, genetics, age, and ethnicity [1]. Clinical treatments of cancer are available, which include hormonal therapy, chemotherapy, radiation therapy, and chemotherapie drugs [2]. Natural chemotherapy drugs from herbal plants, which may induce apoptosis in cancer cells, can serve as chemotherapeutic agents [3]. Natural compounds that have high antioxidants have been demonstrated to have a protective effect against cancer development [4]. Numerous scientific studies have been conducted to explore the antiproliferative effect of OS. Sahib et al [6] discovered that the methanolic extract of OS could

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