Validated high‐performance thin‐layer chromatographic method for the quantification of thymoquinone in <i>Nigella Sativa</i> extracts and formulations

Velho‐Pereira, Reelma; Barhate, Chandrashekhar R.; Kulkarni, Sudheendra; Jagtap, Aarti

doi:10.1002/pca.1289

Cited by 29 publications

(26 citation statements)

References 13 publications

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“…However in 2014, Salmani et al reported that the UV-vis spectra of thymoquinone can be characterized by a prominent peak around 254-257 nm. Thymoquinone, thymol, thymohydroquinone, dithymoquinone and many others are among the active compounds isolated from the seed of N. sativa which responsible in providing the therapeutic properties (Zribi et al, 2014). Hydroquinone is described to give another peak around 290 nm but no distinguished peak is observed at 294 nm, although the same wavelength was used in other study to detect thymoquinone (Salmani et al, 2014).…”

Section: Introductionmentioning

confidence: 94%

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Method Development and Validation using UV Spectrophotometry for Nigella sativa Oil Microparticles Quantification

Ismail¹,

Doolaanea²,

Mohamed³

et al. 2015

J App Pharm Sci

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Nigella sativa oil (NSO) has been exploited for medical purposes for many generations. The fabrication of microparticles containing NSO intended for sustained release was done to be used in treating osteomyelitis. Method in quantifying NSO using UV-spectroscopy was developed and validated. Linearity shown a good correlation coefficient with the values higher than 0.995, both for actual and different analysts. The LOD and LOQ values were recorded to be 2.89 µg/mL and 8.75 µg/mL respectively. In addition, the highest %RSD values for the intermediate and repeatability studies were 0.970% and 0.445% which suggested the method was precise. The percentage recovery for 4 known concentrations gave the range between 98.16% to 99.39%, indicating the high accuracy of the method. The parameters analyzed in this study were in accordance with ICH Q2 (R1) guidelines.

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Section: Introductionmentioning

confidence: 94%

“…HPLC and GC-MS in quantifying the N. sativa seeds extract (Velho et al, 2011;Avula et al, 2010;Lupidi et al, 2010). However, these two common quantifying methods are relatively expensive and time consuming.…”

Section: Introductionmentioning

confidence: 99%

Method Development and Validation using UV Spectrophotometry for Nigella sativa Oil Microparticles Quantification

Ismail¹,

Doolaanea²,

Mohamed³

et al. 2015

J App Pharm Sci

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“…It produces capsule-like fruit full of numerous white trigonal seeds which turn black upon exposure to air after the fruit capsule matures and opens [41]. The black seeds also called black cumin, or black caraway seed are the richest source of its bioactive compounds including thymoquinone (TQ; 2-isopropyl- 5-methyl-1, 4-benzoquinone) [31, 36, 42]. Thymoquinone (TQ) is the most abundant bioactive component in the volatile oil from Nigella sativa black seeds [43, 44]; ranging from 18.4–24.0% [44–47] to as high as 27.8 – 57% TQ [31, 36, 37, 42, 48, 49] depending on where the plant is cultivated and how the oil is extracted [39, 50].…”

Section: Introductionmentioning

confidence: 99%

“…The oil and TQ have also shown potent anti-inflammatory effects on several inflammation-based models including experimental encephalomyelitis, colitis, peritonitis, asthma, and arthritis through suppression of pro-inflammatory mediators [31, 37, 46, 50, 54, 58, 59]. Additionally, studies show that TQ possess neuroprotective [46, 47, 60–62], antimicrobial [42, 50], antidiabetic [42, 50, 63], anticancer [34] and beneficial immunomodulatory properties [36, 44, 54]. …”

Section: Introductionmentioning

confidence: 99%

The Antioxidant Effects of Thymoquinone in Activated BV-2 Murine Microglial Cells

et al. 2016

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Both neuroinflammation and microglial activation are pathological markers of a number of central nervous system (CNS) diseases. During chronic activation of the microglial cells, the induced release of excessive amounts of reactive oxygen species (ROS) and pro-inflammatory cytokines have been implicated in several neurodegenerative diseases such as Alzheimer’s disease. Thymoquinone (TQ), a major bioactive compound of the natural product Nigella sativa seed, has been shown to be effective against numerous oxidative stress-induced and inflammatory disorders as well as possess neuroprotective properties. In this study, we investigated the antioxidant effects of TQ on LPS/IFNγ or H2O2-activated BV-2 microglia by assessing the levels of specific oxidative stress markers, the activities of selected antioxidant enzymes, as well as profiling 84 key genes related to oxidative stress via real-time reverse transcription (RT2) PCR array. Our results showed that in the LPS/IFNγ-activated microglia TQ significantly decreased the cellular production of both superoxide and nitric oxide 4-fold (p<0.0001) and 6 fold (p<0.0001), respectfully. In the H2O2-activated microglia, TQ also significantly decreased the cellular production of superoxide 3-fold (p<0.0001) and significantly decreased hydrogen peroxide levels ~20% (p<0.05). Moreover, TQ treatment significantly decreased the levels oxidative stress in the activated BV-2 as evidenced by the assessed levels of lipid hydroperoxides and glutathione. TQ significantly decreased the levels of lipid hydroperoxides 2-fold (p<0.0001) and significantly increased the levels of antioxidant glutathione 2.5-fold (p<0.0001) in the LPS/IFNγ-activated BV-2 cells. In the H2O2-activated microglia, TQ significantly decreased lipid hydroperoxides 8-fold (p<0.0001) and significantly increased glutathione 15% (p<0.05). Activities of antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), in the TQ-treated microglial cells also reflected a reduced oxidative stress status in the cellular environment. SOD and CAT activities were 6 fold (p<0.0001) and 5 fold (p<0.0001) lower, respectfully, for the LPS/INFγ-activated microglia treated with TQ in comparison to those that were not. For the H2O2-activated microglia treated with TQ, SOD and CAT activities were 5 fold (p<0.0001) and 3 fold (p<0.01) lower, respectfully, compared to the untreated. Furthermore, RT2 PCR array profiling of the selected 84 genes related to oxidative stress confirmed that TQ treatment in the LPS/IFNγ-activated microglia downregulates specific pro-oxidant genes, upregulates specific anti-oxidant genes, and enhances the up- or downregulation of specific genes related to the cells’ natural antioxidant defense against LPS/IFNγ activation. These findings suggest that TQ may be utilized as an effective therapeutic agent for delaying the onset and/or slowing/preventing the progression of microglia-derived neurodegeneration propagated by excessive oxidative stress in the CNS.

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“…The presence of volatile components was confirmed by GC/MS spectroscopy whereas UV/Vis spectral analysis was performed for presence of TQ. The total phenolic content in MNS was quantified by Folin-Ciocalteu procedure[18], the percentage saponins (mg/100 mg) was estimated by anisealdehyde-sulphuric acid method[19] while the quantification of TQ was done by reported HPTLC method[20]. The IC 50 values obtained for MNS extracts for inhibition of free radicals was determined by free radical (2,2-diphenyl-1-picrylhydrazyl (DPPH)) scavenging activity[21] whereas the IC 50 value for inhibition of superoxide radical was performed by standard reported methods[22].…”

Section: Methodsmentioning

confidence: 99%

Radioprotection by macerated extract of Nigella sativa in normal tissues of fibrosarcoma bearing mice

et al. 2012

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The current study was undertaken to study the effect of a macerated extract of Nigella sativa seeds in normal as well as in tumour bearing mice against gamma radiation-induced cellular damage to normal tissues. This was done to mimic the clinical setting where in, normal tissues of cancer patients undergoing radiotherapy are exposed to the deleterious effects of radiation. The protection of cellular DNA was analysed in peripheral blood leucocytes of whole body irradiated mice following pretreatment with macerated extract of Nigella sativa seeds (100 mg/kg), using alkaline comet assay, and also estimating biochemical and blood parameters such as levels of antioxidant enzymes superoxide dismutase and catalase, thiobarbituric acid reactive substances and protein oxidation in organs such as spleen, liver, brain and intestine haemoglobin and total leucocyte count, respectively. The results showed that the macerated extract of Nigella sativa seeds protected the liver, spleen, brain and intestines both in normal as well as tumour bearing mice. This study concludes that macerated extract of Nigella sativa seeds has protective effects against radiation-induced damage and biochemical alterations which could be attributed to the ability to scavenge free radicals and its antioxidant properties. Hence macerated extract of Nigella sativa seeds, could be used in combination with radiation to protect against oxidative stress in normal tissues and improving the quality of life of cancer patients by mitigating unwanted side effects of radiation in normal tissues.

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Validated high‐performance thin‐layer chromatographic method for the quantification of thymoquinone in Nigella Sativa extracts and formulations

Abstract: The HPTLC method developed was found to be relatively simple, rapid and accurate for the routine analysis of TQ in extracts, marketed oils, polyherbal and in-house formulations.

Cited by 29 publications

References 13 publications

Method Development and Validation using UV Spectrophotometry for Nigella sativa Oil Microparticles Quantification

Method Development and Validation using UV Spectrophotometry for Nigella sativa Oil Microparticles Quantification

The Antioxidant Effects of Thymoquinone in Activated BV-2 Murine Microglial Cells

Radioprotection by macerated extract of Nigella sativa in normal tissues of fibrosarcoma bearing mice

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