Hindawi Journal of Chemistry Volume 2020, Article ID 6749176, 17 pages https://doi.org/10.1155/2020/6749176 Research Article UPLC-ESI-MS/MS Profile and Antioxidant, Cytotoxic, Antidiabetic, and Antiobesity Activities of the Aqueous Extracts of Three Different Hibiscus Species Hanan M. Al-Yousef ,1 Wafaa H. B. Hassan,2 Sahar Abdelaziz,2 Musarat Amina ,1 Rasha Adel,2 and May A. El-Sayed2 1Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia 2Department of Pharmacognosy, Faculty of Pharmacy, Zagazig University, Zagazig, Egypt Correspondence should be addressed to Hanan M. Al-Yousef; [email protected] Received 8 February 2020; Revised 20 May 2020; Accepted 3 June 2020; Published 25 June 2020 Academic Editor: Casimiro Mantell Copyright © 2020 Hanan M. Al-Yousef et al. )is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. )e aqueous extracts of Hibiscus calyphyllus (HcA), Hibiscus micranthus (HmA), and Hibiscus deflersii (HdA) growing in Saudi Arabia did not receive enough attention in phytochemical and biological studies. )is inspired the authors to investigate the phytochemicals of these extracts for the first time using UPLC-ESI-MS/MS in negative and positive ionization modes. )e analysis afforded the tentative identification of 103 compounds including phenolic compounds, flavonoids, and anthocyanins. Moreover, in vitro evaluations of their cytotoxic, antioxidant, antidiabetic, and antiobesity activities were carried out. )e results showed that aqueous extract of Hibiscus calyphyllus had the highest activity as an antioxidant agent (SC50 = 111 ± 1.5 μg/mL) compared with ascorbic acid (SC50 = 14.2 ± 0.5 μg/mL). MTTassay was used to evaluate cytotoxic activity compared to cisplatin. Hibiscus deflersii showed the most potent cytotoxic effect against A-549 (human lung carcinoma) with IC50 = 50 ± 5.1 μg/mL, and Hibiscus micranthus showed a close effect with IC50 = 60.4 ± 1.7 μg/mL. Hibiscus micranthus showed the most potent effect on HCT-116 (human colon carcinoma) with IC50 = 56 ± 1.9 μg/mL compared with cisplatin (IC50 = 7.53 ± 3.8 μg/mL). HcA and HdA extracts showed weak cytotoxic activity against A-549 and HCT-116 cell lines compared to the other extracts. Eventually, Hibiscus deflersii showed astonishing antidiabetic (IC50 = 56 ± 1.9 μg/mL) and antiobesity (IC50 = 95.45 ± 1.9 μg/mL) activities using in vitro α-amylase inhibitory assay (compared with acarbose (IC50 = 34.71 ± 0.7 μg/mL)) and pancreatic lipase inhibitory assay (compared with orlistat (IC50 = 23.8 ± 0.7 μg/mL)), respectively. In conclusion, these findings are regarded as the first vision of the phy- tochemical constituents and biological activities of different Hibiscus aqueous extracts. Hibiscus deflersii aqueous extract might be a hopeful origin of functional constituents with anticancer (on A-549 cell line), antidiabetic, and antiobesity activities. It might be a natural alternative remedy and nutritional policy for diabetes and obesity treatment without negative side effects. Isolation of the bioactive phytochemicals from the aqueous extracts of aerial parts of Hibiscus calyphyllus, Hibiscus micranthus, and Hibiscus deflersii and estimation of their biological effects are recommended in further studies. 1. Introduction Many Hibiscus species are valued as ornamental plants and are cultivated in gardens [2]. Fruits of some Hibiscus (Malvaceae) consists of approximately 200 species species are used as food; a soft drink is provided from widely distributed in tropical and subtropical regions of the flowers of some species (H. sabdariffa L.) and also used in world [1]. Hibiscus is a genus of herbs, shrubs, and trees [2]. food industry, for example, in cakes, wines, syrups, Phytochemical investigation of Hibiscus has been reported jellies, puddings, and cold or hot beverages and as a to contain many classes of secondary metabolites including colorant for herbal teas [1]. Since ancient times, Hibiscus anthocyanins, flavonoids, steroids, terpenoids, alkaloids, has been used in traditional folk medicine for different quinones, and sesquiterpene [2]. disorders. 2 Journal of Chemistry Various pharmacological effects have also been shown Prof. Dr. Mohamed Yousef from the Pharmacognosy De- for Hibiscus and its components such as antihypertensive, partment, College of Pharmacy of King Saud University, and antiatherosclerotic, antioxidant, antihypercholesterolemic, voucher specimens (H. calyphyllus no. HA-234, H. deflersii hypolipidemic, antinociceptive, anti-inflammatory, antipy- no. HA-567, and H. micranthus no. HA-16240) were kept at retic, analgesic, antifungal, antibacterial, antifertility, anti- the herbarium of the department. diabetic, anticancer, antimutagenic, chemopreventive, anthelminthic, and anticonvulsant activities [1, 3, 4]. Hibiscus deflersii (HdA), which is native to Ethiopia and 2.2. Preparation of Extracts. Air-dried powder aerial parts of grown as ornamental plant worldwide, is 1 m high erect selected plant samples (600 g) were individually extracted ° perennial or annual leafy untidy shrub of bright green with distilled water at 100 C with continuously shaking for narrow dentate leaves surrounding bright crimson-red 3 hrs. )e marc of each plant material was extracted thrice flowers. Many interesting pharmacological activities of HdA under similar conditions by repeating the above-mentioned had been reported; its leaves extract is used to treat cardiac procedure. )e aqueous extracts were then filtered by disorders and diabetes. However, its flower infusion and centrifugation, and the filtrates were pooled. )e obtained extract are used as demulcent and emollient, while its de- filtrates were freed from the solvent by freeze-drying to get coction is used for the treatment of bronchial catarrh [5]. dark brown solid masses. )e weight of resulted residues was Hibiscus micranthus (HmA), which is commonly dis- 34.02, 26.25, and 36.41 g for H. calyphyllus (HcA), H. deflersii tributed from south to western part of Saudi Arabia, is a (HdA), and H. micranthus (HmA), respectively. 45 cm shrub with heavy leaves, white flowers, and short pedicels with very distinctive capsules of pea-size fruits and 2.3. Chemicals and Reagents. Sigma Aldrich (Hamburg, is distributed vastly in Saudi Arabia, India, tropical Africa, Germany) provided all the chemical and reagents used and Ceylon. Its flowers and fruits exhibited antidiabetic and throughout the experiments, including HPLC grade meth- laxative activities when used orally, and when applied anol (≥99.9%), acetonitrile, water for ESI-MS analyses, re- topically it is used as antidandruff agent. )e plant also agent grade formic acid (≥95%), 2,2-diphenyl-1- showed anti-inflammatory, antipyretic, antitumor, antimi- picrylhydrazyl (DPPH) (freshly dissolved in methanol at a crobial, and antiviral activities. Literature detects a wide concentration of 0.004%), ascorbic acid (99%), Dulbecco’s range of phytochemicals in HmA as flavonoids, phenolic Modified Eagle’s Medium (DMEM), L-glutamine, dini- acids, fatty alcohols, fatty acids, β-sitosterol, and alkanes [5]. trosalicylic acid (DNS, colour reagent), p-nitrophenyl bu- Hibiscus calyphyllus (HcA) is 1 m high leafy shrub tyrate (NPB, ≥98%), and potassium phosphate buffer (pH characterized by bright yellow flower with dark red center 6.0). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium surrounded by simple wide serrate leaves. It is commonly bromide (MTT), cisplatin, DMSO, porcine pancreatic lipase found in Jazan, south-western region of Saudi Arabia. )e and α-amylase enzyme, acarbose (≥95%), and orlistat ethyl acetate fraction of this plant showed potent antioxidant (≥98%) were used as control. Human lung carcinoma (A- activity [5]. 549) and human colon carcinoma (HCT-116) cell lines were Hibiscus aerial parts are considered as food crops con- obtained from the American Type Culture Collection sumed as hot or cold beverages (aqueous extract). Numerous (ATCC, Rockville, MD). scientific papers have been published discussing the chemical contents of different fractions of HcA, HdA, and HmA, which showed their high biological effectiveness, 2.4. UPLC-ESI-MS/MS. Ultra-performance liquid chroma- having antioxidant, antidiabetic, antiobesity, and cytotoxic tography with electrospray ionization quadrupole-linear ion activities. For our knowledge, the previous literature on trap-tandem mass spectrometry analysis, performed on ESI- Hibiscus species suffers from insufficiency of detailed in- MS positive and negative ion acquisition mode, was carried formation on the phytoconstituents of aqueous extracts of out on a XEVO TQD triple quadruple instrument. Method Saudi HdA, HcA, and HmA and their biological activities. in a multiple-reaction monitoring (MRM) mode was )erefore, the aims of the present research were (i) to employed for the quantitative determination of phyto- perform direct analysis of aqueous extracts, which relies on chemicals. )e crude Hibiscus extracts were analyzed by UPLC coupled with ESI-MS/MS detection, (ii) to detect the UPLC, in order to obtain chromatographic profiles of the antioxidant, antidiabetic, antiobesity, and anticancer activ- more polar portions of the extracts, which contain phenolic ities of aqueous extracts of HdA, HcA, and HmA; and (iii) to and flavonoid compounds. )e samples were dissolved in anticipate the components responsible for antioxidant, HPLC grade methanol, filtered through 0.2 μm membrane