Phytochemical and Safety Evaluations of Volatile Terpenoids from Zingiber cassumunar Roxb. on Mature Carp Peripheral Blood Mononuclear Cells and Embryonic Zebrafish

Abstract

Pharmaceutical products of essential oil from Zingiber cassumunar Roxb. are extensively being developed, while the research on their safety is seldom documented. The aim of the present study was to evaluate the phytochemical profile and the effect of cassumunar ginger oil on cell-based assay and the zebrafish model. The essential oil was isolated from fresh rhizomes of Z. cassumunar using simultaneous steam-distillation. Chemical composition was analyzed using gas chromatograph coupled to a mass spectrometer (GC-MS). Effect of cassumunar ginger oil on adult carp fish peripheral blood mononuclear cells (PBMCs) was investigated using MTT assay. The embryotoxic and teratogenic effects of cassumunar ginger oil were studied in zebrafish embryos. GC-MS results showed that the essential oil was composed of sabinene (43.54%) and terpinen-4-ol (29.52%) as the major phytoconstituents. No fish PBMC cytotoxic effect was observed with the concentration less than 50 µg/mL of cassumunar ginger oil. Our results showed for the first time the embryotoxic and teratogenic effects of cassumunar ginger oil in zebrafish embryos. The result indicated that the cassumunar ginger oil induced zebrafish embryotoxicity in a concentration-dependent manner. At 500 µg/mL of cassumunar ginger oil demonstrated significantly moderated embryotoxicity within 24 h (p < 0.05). The survival rate of 100 µg/mL of cassumunar ginger group was markedly declined to zero at 96-h post-fertilization (log-rank test, p = 0.001). However, survival rates of zebrafish embryo in the 1 and 10 µg/mL cassumunar ginger groups were more than 90% throughout the trial period. Moreover, very low teratogenicity to the zebrafish embryo was also observed in 1 and 10 µg/mL of cassumunar ginger groups. Our findings suggest that there is hardly any cytotoxicity, embryotoxicity and teratogenicity at concentrations less than 10 µg/mL of cassumunar ginger oil. However, the toxicity assessment of its pharmaceutical product should prove for further consumer protection.

Phytochemicals and Bioactivities of Zingiber cassumunar Roxb

“…Pharmaceutical or cosmeceutical products using essential oil from the rhizomes of Z. cassumunar have been developed; thus, the chemical composition of essential oil has been analyzed using usually gas chromatography-mass spectrometry (GC–MS). Monoterpenoids, sabinene ( 58 ), and terpinene-4-ol ( 59 ) were identified as major constituents [ 39 ] in rhizome essential oil, and α-terpinene ( 60 ), and γ-terpinene ( 61 ) were also discovered [ 22 , 40 ]. In addition to these components, the essential oil produced by steam contained approximately 25% phenylbutenoids, and oil extracted with light petroleum contained approximately 46% phenylbutenoids [ 41 ].…”

“…The essential oils included sabinene and terpinen-4-ol as major volatile terpenes [ 22 , 23 , 24 , 25 ]. The extracts and constituents of Z. cassumunar have diverse bioactivities, including antioxidant [ 21 , 26 , 27 , 28 ], anti-inflammatory [ 7 , 9 , 12 , 19 , 23 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 ], anticancer [ 8 , 13 , 15 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 ], neuroprotective/neurotrophic [ 14 , 19 , 20 ], cosmeceutical [ 17 , 21 ], and antifungal/antibacterial [ 22 , 23 , 24 ] activities. Although there has been a review of the clinical effects of various formulations using Plai ( Z. cassumunar ) on were pain relief, acne treatment, and antihistamine [ 1 ], there has been no previous report summarizing the accumulated studies in the literature on the phytochemicals and in vitro and in vivo biological properties of Z. cassumunar , including our previous studies that have contributed to discovering the chemical diversity and biological activities of this plant […”

Phytochemical and Safety Evaluations of Zingiber ottensii Valeton Essential Oil in Zebrafish Embryos and Rats

“…Water was poured out slowly from one corner of the spawning tank, and the fertilized eggs were collected with a plastic pipette and transferred to clean Petri dishes containing embryo water prepared with minor modifications of the previously reported protocol [ 19 ]. The fertilized embryos were carefully washed with sea salt egg water (60 mg/L sea salt and 2 mg/L methylene blue) to remove debris [ 20 ], while unhealthy and dead embryos were removed by aspiration using a plastic pipette. Fertilized embryos were kept at 26.5 °C and allowed to develop for 6 h postfertilization (hpf).…”

“…Dead embryos were examined under a stereomicroscope, and the time–kill analysis was conducted at 24, 48, 72, 96, and 120 hpf. Survival rates and the median survival time were also evaluated [ 20 ].…”

“…Examples of teratogenicity identified included deformities in somite otolith and eyes, failure of tail detachment, absence of heartbeat or blood circulation, yolk sac or pericardial edema, yolk sac malabsorption, and skeletal malformation and delayed hatching [ 23 ]. The malformed embryos and larvae were captured with an Olympus digital camera (OM-D E-M10 Mark III), and the teratogenicity was evaluated by the percentage of embryos or larvae with abnormalities to the number remaining of normal embryos, as previously described [ 20 ].…”