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Assessment of the Antimicrobial and Free Radical Scavenging Activities Of Oriental Pharmacy and Experimental Medicine https://doi.org/10.1007/s13596-018-0316-z RESEARCH ARTICLE Assessment of the antimicrobial and free radical scavenging activities of Moluccella spinosa, Helichrysum sanguineum, and Styrax ofcinalis folkloric medicinal plants from Palestine Nidal Jaradat1 · Motasem Al‑Masri2 · Abdel Naser Zaid1 · Fatima Hussein1 · Khalid Ahmad Shadid3 · Fuad Al‑Rimawi4 · Khaled Shayeb1 · Afnan Sbeih1 · Alia Eid1 Received: 18 July 2017 / Accepted: 20 April 2018 © Institute of Korean Medicine, Kyung Hee University and Springer Science+Business Media B.V., part of Springer Nature 2018 Abstract The emergence of pathogenic microbes with increased resistance to established antibiotics provides a major incentive for the discovery of new antimicrobial agents. Herbals may provide valuable solutions for this global problem. In addition, the replacement of harmful synthetic antioxidants with natural ones may prevent various serious diseases. The present investigation describes for the frst time the antioxidant and antimicrobial activities of the aqueous and organic extracts of Helichrysum sanguineum, Moluccella spinosa and Styrax ofcinalis plants aerial parts. The free radical scavenging activ- ity was estimated using the 2,2-diphenyl-1-picrylhydrazyl method, while the antimicrobial activity was evaluated against selected microbial strains from American Type Culture Collection and clinical isolates such as Shigella sonnie, Staphylococ- cus aureus, Enterococcuss feacium, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Epidermophyton foc- cosum and Methicillin Resistant Staphylococcus aureus (MRSA) using minimum inhibitory concentration assay. A mixture of phytochemical compounds was found in all of the studied plants extracts which also showed remarkable potentials of antioxidant and antimicrobial activities. The current study provides initial data that justify the use and importance of these plants in the Palestinian traditional medicine. In addition, it provides evidence that the aqueous and organic extracts of H. sanguineum, M. spinosa and S. ofcinalis exhibited interesting antioxidant activity comparing with Trolox. Furthermore, the organic extract of H. sanguineum strongly exhibited bacterial growth of S. aureus, E. faecium and MRSA which suggested to be used as antibiotic alternative or as sufcient natural food preservative. Keywords Helichrysum sanguineum · Moluccella spinosa · Styrax ofcinalis · Antibacterial · Antifungal · Antioxidant Introduction The use of plants as natural free radical scavenging agents in * medicine and in the processed foods have become of increas- Nidal Jaradat ing importance in the pharmaceutical and food industries [email protected] as natural and safe alternatives to synthetic antioxidants. 1 Department of Pharmacy, Faculty of Medicine and Health However, these natural products are claimed for their protec- Sciences, An-Najah National University, P. O. Box 7, tive efects against cardiovascular diseases, certain forms of Nablus, Palestine cancer and photosensitivity reactions (Grifths et al. 2016; 2 Department of Biology and Biotechnology, Faculty Zamora and Hidalgo 2016). In addition, antibiotic resistance of Science, An-Najah National University, P. O. Box 7, has become a serious global concern, and the discovery of Nablus, Palestine 3 novel antimicrobial natural products may provide a valu- Department of Chemistry, Faculty of Science, able solution to overcome this problem (Laxminarayan et al. Islamic University in Madinah, P. O. Box 41433, Almadinah Almonawarah, Saudi Arabia 2013). Herbals can be used to solve these problems, and the tra- 4 Department of Chemistry, Faculty of Science and Technology, Al-Quds University, P. O. Box 20002, ditional medicine is rich with herbal remedies which treated East Jerusalem, Palestine Vol.:(0123456789)1 3 N. Jaradat et al. various infectious diseases and cure the oxidative damages rock technology, BT85, China), sterile syringe flter hydro- caused by free radicals in human body. phobic, 0.45 µm 25 mm (PTFE, China), weighing scale Moluccella spinosa L., Helichrysum sanguineum (L.) (Adam Equipments, USA), incubator with CO2 (Tuttnauer, Kostel, and Styrax ofcinalis L. are folkloric medicinal Jerusalem), balance (Rad wag, AS220/c/2, Poland), multi- plants from Palestine and used in the treatment of some channel micro-pipet (Eppendorf, Germany) and multichan- urological infectious diseases and oxidative stress (Viegas nel micro-pipet (Thermo Fisher Scientifc, India). et al. 2014; Jaradat 2005). However, H. sanguineum (Compositae family) is com- Chemicals monly known as Red Everlasting, and Blood of the Mes- siah. It is used in the folk medicine for treatment of vari- Ethanol, sodium hydroxide, n-hexane, and acetic acid were ous urological infectious diseases and wildly distributed in obtained from Lobachemie, India, and Iron (III) chloride, Palestine, Jordan, Lebanon, Turkey and Syria and (Aslana Millon’s reagent, Benedict’s reagent, Dimethyl sulfoxide et al. 2006; Erolu et al. 2010; Mericli et al. 1984). The (DMSO) were purchased from Riedeldehan, Germany. leaves, stems and fowering heads of H. sanguineum contain While Molish’s reagent, sulfric acid, and iodine solution kaempferol, 5,7-dihydroxy-3,8-dimethoxy favone, 3,5,7-tri- were obtained from Alfa-Aesar, England. Also, chloroform hydroxy-6,8-dimethoxy favone, kaempferol 3-glucoside, and HCl were purchased from Sigma-Aldrich, Germany. In quercetin 4′-glucoside favonoids, pelargonidin 3-glucoside addition, Magnesium ribbon and Ninhydrin solution were and peonidin 3-glucoside anthocyanins (Mericli et al. 1984). obtained from Alfa Agar, England. Moreover, the nutrient While, M. spinosa plant belongs to the Lamiaceae family broth and MacConkey agar were brought from Himedia and commonly known as Spiny molucca. It is wildly distrib- (India), while BBL Mueller–Hinton II broth, Difco Sab- uted in Palestine and other Mediterranean regions, especially ouraud Dextrose Agar, and Bacto tryptic soy broth were in the roadsides and fallow felds areas (Ronel et al. 2010). obtained from Dickinson and company sparks (USA). The main chemical composition of M. spinosa aerial parts essential oil collected in Sicily were α-pinene, caryophyllene Plants materials oxide and β-caryophyllene. This oil showed slight antibac- terial and a moderate antifungal activities (Casiglia et al. The aerial parts of H. sanguineum, M. spinosa and S. ofc- 2015). inalis plants were collected in May 2016 from Tulkarem In addition, the S. ofcinalis (family Styracaceae) used in region of the West-Bank Area/Palestine. The taxanomical the traditional medicine for treatment of urinary tract infec- identifcations were established by the pharmacognosist Dr. tious diseases, respiratory system infections, and dermatitis. Nidal Jaradat in the Pharmacognosy Laboratory, Faculty It is native to southern regions of Europe and wildly distrib- of Medicine and Health Sciences, An-Najah National Uni- uted in the forests of Palestine (Jaradat 2005; Danin 2004; versity. While, the voucher specimen codes were Pharm- Ali-Shtayeh et al. 2013). However, the leaves major chemi- PCT-1170, Pharm-PCT-1598 and Pharm-PCT-2365, cal composition of essential oil were (E)-2-hexenal, octanol respectively. and geraniol (Tayoub et al. 2006). The used plants parts were washed well and dried in the The current investigation aimed to characterize the phy- shade at room temperature (25 ± 2 °C), and humidity (55 ± 5 tochemical components also to evaluate the antioxidant and RH). After drying, the used plant materials were grounded antimicrobial efects of H. sanguineum, M. spinosa and S. well using a mechanical grinder and transferred into airtight ofcinalis plants aqueous and organic extracts. These plants containers with proper labeling for future use. were chosen due to their importance in the Palestinian tra- ditional medicine. In addition, to the best of author’s knowl- Preparation of plants extracts edge, the literature survey could not ascertain any antimicro- bial and antioxidant studies carried out on the aerial parts A total of 25 g of the powdered plant material was weighed of these plants. and then extracted by adding 100 ml of n-hexane and 150 ml of 50% ethanol in triple distilled water. The mixture was then shaken for 48 h at room temperature using shaker device Materials and methods that was set at 200 rpm. Afterwards, the mixture was fltered using suction fask and Buchner funnel. The obtained fl- Instrumentation trate was separated individually by separatory funnel into 2 phases; a lower aqueous phase representing the frst aqueous Shaker incubator (Memmert, Germany), rotary evaporator extract and the upper organic phase representing the organic (Heidolph, Germany), stir-mixer (Tuttnauer, Jerusalem), extract. The remaining solid materials were re-extracted sep- grinder (Moulinex model, Uno, China), lyophilizers (Mill arately by 150 ml of 50% ethanol in triple distilled-water and 1 3 Assessment of the antimicrobial and free radical scavenging activities of Moluccella spinosa,… the re-extraction process was carried out as described above. and the Trolox standard were calculated using the following The aqueous extract was dried using freeze dryer for 48 h. formula: Meanwhile, organic extracts were placed in hood at 25 °C Percentage of inhibition of DPPH activity (%) = (B − A)∕B × 100% to evaporate leftover organic solvents and completely dried B organic extracts till use (Jaradat et al. 2014). where = Absorbance value of the control (prepared
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