GC-FID Methods Research Articles

Spermacoce alata Aubl. Essential Oil: Chemical Composition, In Vitro Antioxidant Activity, and Inhibitory Effects of Acetylcholinesterase, α-Glucosidase and β-Lactamase.

Spermacoce alata Aubl. is widely available in the market as traditional Chinese medicine and animal feed, due to its properties of clearing heat and treating malaria and its high-protein and crude fiber content. In this study, the essential oil of S. alata was obtained through hydrodistillation. GC-MS and GC-FID methods were used to identify the chemical components and their relative abundance. Furthermore, the antioxidant capacity was measured using DPPH, ABTS, and FRAP assays, and the inhibitory effects of acetylcholinesterase, α-glucosidase, and β-lactamase were also evaluated. A total of 67 compounds were identified, with the major constituents being palmitic acid (30.74%), linoleic acid (16.13%), and phenylheptatriyne (8.07%). The essential oil exhibited moderate antioxidant activity against DPPH (IC50 > 10 mg/mL), while the IC50 value for the ABTS assay was 3.84 ± 2.12 mg/mL and the FRAP assay value was 87.22 ± 12.22 µM/g. Additionally, the essential oil showed moderate anti-acetylcholinesterase activity (IC50 = 286.0 ± 79.04 μg/mL), significant anti-α-glucosidase activity (IC50 = 174.7 ± 13.12 μg/mL), and potent anti-β-lactamase activity (IC50 = 37.56 ± 3.48 μg/mL). The results suggest that S. alata has the potential for application in pharmacology, warranting further exploration and investigation.

Fermentation Kinetics, Potability, and Sensory Compounds of Joubidwi: An Ethnic Alcoholic Beverage

Background: Fermentation is a metabolic pathway of the breakdown of carbohydrate molecules like glucose, fructose to ethanol and various volatile and non-volatile congeners. The production of alcoholic beverages from foodstuffs like rice, cereals, fruits, maize etc., is an age-old traditional protocol. In this study, we adapted the traditional protocol of preparation of the ethnic beverage Joubidwi, with a fermentation time of 6 days, i.e., aliquots from the fermented mash (cooked rice) were collected on a daily basis from day 1 to day 6. Objectives: This study aimed to get insight into fermentation kinetics, ethanol content, carbohydrate content, antioxidant activities, bitterness, protein content, pH, colour, and turbidity of the ethnic alcoholic beverage Joubidwi and to identify the sensory compounds and polyphenol molecules. Method: UV-Visible spectroscopic methods, GC-FID and GC-MS analytics methods were used for the estimation and identification of carbohydrates, proteins, ethanol contents, and sensory compounds, respectively. Reverse phase HPLC was adapted for the identification of polyphenol molecules in Joubidwi. Results: A pseudo-first-order fermentation kinetics was observed from day 1 to day 6 fermentation of Joubidwi. Apart from ethanol content (60.48 ± 1.36 mg/mL after 72 h fermentation to 78.80 ± 1.48 mg/mL after 144 h fermentation), the beverage had nutritional values and good antioxidant properties with 22.38 to 35.11% Radical Scavenging Effect (RSE) from 1 to 6th day of fermented Joubidwi. The beverage also contained 12 sensory compounds. Gallic acid was identified as polyphenol in Joubidwi samples. Conclusion: Joubidwi is an alcoholic beverage prepared by Bodos using rice and certain herbs through a traditional fermentation process. The fermentation process follows pseudo-first-order kinetics. The beverage is potable.

A validated comparative study of RP-HPLC, GC-FID and UV spectrophotometric methods for the quantification of eugenol isolated from Syzygium Aromaticum L

A validated comparative study of RP-HPLC, GC-FID and UV spectrophotometric methods for the quantification of eugenol isolated from Syzygium Aromaticum L

Greening of the method for simultaneous determining the enisamium iodide and tilorone dihydrochloride using GC-FID assay

Pharmaceutical companies in Ukraine aspire to develop their innovative medicinal products and successfully introduce them to the global market. However, along with the prospects of increased usage of these pharmaceuticals, there arises a challenge of heightened waste production, making them a part of the over twenty million tons of PPCPs produced annually. Consequently, one of the tasks in producing new pharmaceuticals is the development of methodologies and approaches not only for quality control but also for their determination in the environment matrices.
 The aim. Develop and validate GC-FID chromatographic method for the simultaneous determination of Enisamium iodide and Tilorone dihydrochloride, evaluate their applicability, and compare their "greenness" with the previously developed HPLC method.
 Materials and methods. The determination of the Tilorone dihydrochloride and Enisamium iodide was carried out by gas chromatography with a flame ionization detector using the Rxi-5 ms (30 m long, 0.25 mm outer diameter and 0.25 μm liquid stationary phase thickness)
 Results. Chromatographic GC-FID methods have been developed for the simultaneous determination of Enisamium iodide and Tilorone dihydrochloride. Optimal sample preparation conditions were established, and a validation process was conducted. A comparison with the previously developed HPLC method was made regarding "greenness."
 Conclusions. The developed GC-FID methodology is accurate and more environmentally friendly compared to the previously established methods. It can be recommended to determine Enisamium iodide and Tilorone dihydrochloride in the environmental matrices. It is considered environmentally friendly based on the overall GREENness (AGREE) scale, scoring 0.73 (>0.70), which demonstrates the environmentally favourable nature of the proposed analytical approach

Development of a Pure Certified Reference Material of D-Mannitol.

A new certified reference material (CRM) of D-mannitol (GBW(E) 100681) has been developed in this study. We describe the preparation, structure determination, characterization, homogeneity study, stability study, as well as uncertainty estimation. The main component was 99.91% ± 0.01%. The moisture content of the candidate CRM was 0.036% ± 0.002%, as measured by Karl Fischer titration. The nonvolatile and volatile impurities in the candidate CRM were all much less than 0.01%, which was determined by the ICP-MS and headspace GC-FID methods, respectively. The purity of the D-mannitol CRM was 99.9% ± 1.1% (k = 2), as measured by the two independent approaches involving the mass balance method (MB) and quantitative nuclear magnetic resonance technique (qNMR). The D-mannitol CRM was stable during the monitoring period for each temperature. It is stable for up to 48 months at room temperature and 28 days at 50 °C. The uncertainty was evaluated by combining the contributions from characterization, homogeneity, and stability. The developed D-mannitol CRM would effectively support method validation and proficiency testing, as well as effectively guarantee the accuracy, reliability, and comparability of results.

GC-FID and GC-MS detection of Ethyl Isobutyryl Acetate as regulatory starting material

We developed GC-FID and GC-MS methods to determine the assay and chromatographic purity of Ethyl isobutyryl acetate (EIBA). The symmetric peak shape was on a DB-Wax stationary phase with the dimensions of 30 m column length, 0.25 mm as internal diameter, 0.50 microns film thickness with the mobile phase as helium gas in temperature ramping mode at a flow rate of 1.5 mL/min using flame ionization detector and mass spectrometer. The USP resolution between EIBA and its unknown impurities was more than 2.0, indicating a significant separation. Regression analysis shows a correlation coefficient greater than 0.999 for EIBA assay and chromatographic purity techniques. The detection and quantitation limits of EIBA are 0.01% and 0.03% for the chromatographic purity procedure. This method indicates that the recovery at different levels is 90 to 110% accurate. The test solution was stable in the diluent for 48 h and subjected to stress conditions. The mass balance was close to 99.5%.

Chemical composition, nutritional values, elemental analysis and biological properties of Prunus mahaleb L.: From waste to new potential sources for food, cosmetic and drug industry

Prunus mahaleb L. seeds are widely used in the baking industry as spice for flavouring. While achieving the mahaleb powder for commercial use in local companies, the seeds are sometimes mixed unintentionally with their kernels (stony shell) during sieving procedure. As the chemical and biological properties of the kernel of seeds are not well known, the kernel, seeds and the market sample obtained from a local supplier are decided to investigate individually. The antioxidant capacity and the trace element analyses of the seeds and kernels were performed by TEAC and CUPRAC assays and ICP-MS. The market sample was analysed for nutrient and oil composition. Fatty acid composition of the oil was determined with GC-FID and GC-MS methods. The chemical composition of the methanol extract of the seeds and kernels was studied by LC-MS/MS. Trace elements Mn, Co, Ni, Cu, Zn and Se were determined in the methanol extracts, ethyl acetate and water fractions. Mushroom tyrosinase enzyme inhibitions of 33 different extracts and fractions of various waste parts of the plant (kernel, fruit, twigs, flos, leaves) were investigated. The nutritional values showed that the market sample had high nutritious properties. The fatty acid content of the oil was found to be valuable in remedying dysfunctions of the body. Kernel ethylacetate fraction showed the highest tyrosinase enzyme inhibition. The role of copper element on enzyme activity was determined. The kernels are accepted as a waste material may also be evaluated as a good promising choice for food, cosmetic and drug industry.

Proposals for the improving of the existing GC-FID methods for determination of methanol and volatile compounds in alcoholic beverages

Proposals for the improving of the existing GC-FID methods for determination of methanol and volatile compounds in alcoholic beverages

Determination of Major, Minor and Chiral Components as Quality and Authenticity Markers of Rosa damascena Oil by GC-FID.

Rose oil is traditionally produced by the water distillation of Rosa damascena and is of high economic value due to the low essential oil yield. It is therefore a common target for adulteration, which can cause harm to consumers. Current standards for authenticity control only consider the analysis of major components and overlook minor quality markers as well as the enantiomeric ratio of terpenes, which have proven useful in originality determination. The aim of this study was the development of two analytical GC-FID methods for the analysis of 21 and 29 rose oil analytes including major, minor and chiral components on a DB-wax and BGB 178 30% CD (chiral) capillary column, respectively. The total run time for both methods was within 60 min. For all target analytes, the % bias at the lower and upper calibration range varied from -7.8 to 13.2% and -13.1 to 5.2% analysed on the DB-wax column and 0.5 to 13.3% and -6.9 to 7.0% analysed on the chiral column. The chiral analysis successfully separated the enantiomers (+/-)-camphene, (+/-)-rose oxide, (+/-)-linalool, (+/-)-citronellol and (+/-)-citronellyl acetate, as well as the diastereomers of citral and β-damascenone. Both methods were applied to the analysis of 10 authentic rose oil samples and the enantiomeric/diastereomeric ratios, as well as the content of major and minor components, were determined. The identity of the analysed components in the authentic samples was further confirmed by GC-MS.

Chemical Composition, Antitumor Potential, and Impact on Redox Homeostasis of the Essential Oils of Orlaya grandiflora from Two Climate Localities

It is well known that abiotic components can affect biosynthetic pathways in the production of certain volatile compounds. The aim of this study was to compare the chemical composition of essential oils obtained from Orlaya grandiflora (L.) Hoffm. collected from two localities in Serbia (continental climate, OG1) and Montenegro (Mediterranean climate, OG2) and to assess their antitumor potential on the human colon cancer HCT-116 and breast cancer MDA-MB-231 cell lines. EOs obtained by hydrodistillation were analyzed using GC-MS and GC-FID methods. The results indicate considerable differences in the chemical compositions of the two samples. Although in both samples the main class of volatiles observed was sesquiterpenes (47.5% for OG1 and 70.1% for OG2), the OG1 sample was characterized by a high amount of monoterpene hydrocarbons (29.3%), and sesquiterpene germacrene D (29.5%) as the most abundant compound. On the other hand, the OG2 sample contained a high quantity of oxygenated sesquiterpenes (20.6%), and β-elemene (22.7%) was the major constituent. The possible antitumor mechanisms of these EOs in the HCT-116 and MDA-MB-231 cell lines were examined by means of cell viability, apoptosis, redox potential, and migratory capacity. The antiviability potential appeared to be dose dependent, since the results showed that both EOs decreased the viability of the tested cells. Stronger antitumor effects were shown in MDA-MB-231 cells after short-term treatment, especially at the highest applied concentration, where the percentage of viability was reduced by over 40%. All tested concentrations of EOs exhibited proapoptotic activity and elevated activity of caspase-3 in a dose- and time-dependent manner. The results also showed decreased concentrations of superoxide anion radical in the treated cells, which indicates their significant antioxidative role. Long-term treatments showed mild recovery effects on cell viability in both cell lines, probably caused by the balancing of redox homeostasis. Elevated levels of nitrites indicate high levels of nitric oxide (NO) production and suggest its higher bioavailability due to the antioxidative environment. The tested EOs also induced a drop in migratory capacity, especially after short-time treatments. Taken together, these results suggest considerable antitumor activity of both EOs, which could have potential therapeutic applications.

Introducing a Modern Chemotherapeutic Drug Formulated by Etlingera velutina Essential Oil for the Treatment of Human Gastric Cancer

The essential oils from the stem of Etlingera velutina was obtained by a Clevenger-apparatus and the chemical composition of the volatile oil was identified using GC-FID and GC-MS methods. the oil consists of sesquiterpene hydrocarbon. α-Cubebene (29.98%) and 2-undecanone (20.04%) were the main components for the rhizome oil. In the cellular and molecular part of this study, the MTT assay was run to treat of the cell lines by EO for evaluation of the cytotoxicity and anti-human gastric cancer properties of EO. The results revealed a dose-dependently on the viability of malignant gastric cell line. The IC50 of EO were 376 and 371 μg/mL against NCI-N87 and MKN45 cell lines, respectively. For antioxidant analysis, the IC50 of 192 μg/mL was obtained for radical scavenging activity of EO against DPPH.

Harmful volatile substances in recorded and unrecorded fruit spirits

The alcohol-attributable mortality is an important issue in South-Eastern Europe, whereby consumption of homemade spirits is stated to be one of the leading causes. Presence of harmful volatiles in spirits in Serbia was investigated in 26 recorded and 127 unrecorded fruit spirits, collected in 2020 and analysed using GC-FID methods. Statistical analysis confirmed higher content of ethanol, ethyl acetate, n-propanol and iso-butanol in unrecorded spirits. Regarding concentration limits proposed by the Alcohol Measures for Public Health Research Alliance, the one for methanol was exceeded in 4% of recorded and 17% of unrecorded spirits, for acetaldehyde in 4% of unrecorded and for higher alcohols only in one recorded spirit. Risk assessment, conducted using a margin of exposure approach, showed that spirit consumption, even at average level, posed a risk due to their high ethanol content. Regarding acetaldehyde, for males the risk was indicated starting from the regular drinkers only scenario (mean MOE 9937 for recorded and 9123 for unrecorded spirits) and for females who were chronic heavy drinkers of recorded (2324) or unrecorded spirits (1644). Chronic heavy drinkers of both sexes were in the risk of methanol and higher alcohols in case of exclusive consumption of unrecorded spirits.

The Occurrence of Crude Oil-Degrading Bacteria in Some Sponges Collected at the Persian Gulf: Ecological Importance and Biotechnological Application

The Persian Gulf is the most critical aquatic environment in the world. The relationship between marine animals and degrading bacteria not well understand. The purpose of this investigation is the description of symbiont crude oil-degrading bacteria in some sponges in the Persian Gulf. Marine sponges were collected from four stations at the Persian Gulf. Symbiont crude oil-degrading bacteria were isolated from marine sponges by enrichment in ONR7a medium after four subcultures. The isolated bacteria were identified by biochemical and molecular methods. The degradation of crude oil was determined by spectrophotometric, gravimetric, and GC-FID methods for each strain. In this study, 38 crude oil-degrading bacteria were isolated from 8 sponge samples. The efficient strains that had the highest crude oil degradation belonged to the genus Cellulosimicrobium, Shewanella, Alcanivorax and Cobetia. The GC-FID analysis confirmed that the strains IAUK3532, IAUK3537 and IAUK3519 exhibit maximum degradation of crude oil (respectively, 94.5%, 88.13% and 85%). It is observed that when the concentration of crude oil increased, the degradation decreased. It can be concluded from these results that the microbes that coexist in the sponges located at the Persian Gulf can degrade crude oil. These symbionts crude oil-degrading bacteria can be applied to decrease the oil pollution in the Persian Gulf. Also, the symbiotic relationship between sponges and crude oil-degrading bacteria increases sponge survival and contributes to the development of their colonies in the Persian Gulf.

Gloeothece sp.-Exploiting a New Source of Antioxidant, Anti-Inflammatory, and Antitumor Agents.

Bioactive lipidic compounds of microalgae, such as polyunsaturated fatty acids (PUFA) and carotenoids, can avoid or treat oxidation-associated conditions and diseases like inflammation or cancer. This study aimed to assess the bioactive potential of lipidic extracts obtained from Gloeothece sp.–using Generally Recognized as Safe (GRAS) solvents like ethanol, acetone, hexane:isopropanol (3:2) (HI) and ethyl lactate. The bioactive potential of extracts was assessed in terms of antioxidant (ABTS•+, DPPH•, •NO and O2•assays), anti-inflammatory (HRBC membrane stabilization and Cox-2 screening assay), and antitumor capacity (death by TUNEL, and anti-proliferative by BrdU incorporation assay in AGS cancer cells); while its composition was characterized in terms of carotenoids and fatty acids, by HPLC-DAD and GC-FID methods, respectively. Results revealed a chemopreventive potential of the HI extract owing to its ability to: (I) scavenge -NO• radical (IC50, 1258 ± 0.353 µg·mL−1); (II) inhibit 50% of COX-2 expression at 130.2 ± 7.4 µg·mL−1; (III) protect 61.6 ± 9.2% of lysosomes from heat damage, and (IV) induce AGS cell death by 4.2-fold and avoid its proliferation up to 40% in a concentration of 23.2 ± 1.9 µg·mL−1. Hence, Gloeothece sp. extracts, namely HI, were revealed to have the potential to be used for nutraceutical purposes.

Optimization of the Essential Oil Extraction Process from Dong Van Marjoram (E. winitiana var. dongvanensis Phuong.) by Using Microwave Assisted Hydrodistillation, and the Bioactivities of the Oil Against Some Cancer Cell Lines and Bacteria

This study reports on the optimization of a microwave-assisted distillation process to obtain Dong Van marjoram essential oil, and the determination of its composition, content of constituents, and cytotoxic and antimicrobial activities. Using the response surface method (RSM), the optimal essential oil distillation conditions were determined as material size 0.74 (cm), water to material ratio 4.14:1 (mL/g), microwave power 302.4 (W), and distillation time 2.1 hours. At optimal conditions, the mass of Dong Van marjoram essential oil obtained was 0.887 ± 0.007 g, corresponding to a content of 0.6% essential oil in the material. GC-MS and GC-FID methods showed that the main chemical constituents of Dong Van marjoram essential oils obtained were rosefuran epoxide (44.9%), caryophyllene (10.8%), germacrene D (2.6%), and α-humulene (1.3%). The essential oil exhibited moderate inhibition against both tested cancer cell lines, with IC50 values of 23.9 µg/mL (for PC3) and 56.2 µg/mL (for A549). However, the oil exhibited strong effectiveness against three bacterial strains, Escherichia coli, Bacillus subtilis, and Staphylococcus aureus, and a yeast strain, Saccharomyces cerevisiae, with minimal inhibitory concentration (MIC) values ranged from 50 to 100 µg/mL.

A Review: Effects of air, water and land dumpsite on human health and analytical methods for determination of pollutants

Environment pollutants are found here and there in developing countries and these contaminations affect the environment adversely. A few remediation of pollution which includes incineration which is the waste treatment process that involves the combustion of organic substances contained in waste material, the waste handling practices, the recycling resource recovery, the avoidance and reduction methods, the adsorption based on nanotechnology and the bioremediation technology which appears as a cost-effective and environmental friendly approach for cleanup. Recently researches shows that various chemicals (VOCs, BTEX, heavy metals) that might be delivered into the air or water can cause unfriendly health effects which was analyzed based on sample treatments (solid phase extraction: SPE, the liquid-liquid microextraction: LLME, the magnetic solid phase extraction: MSPE) and instruments such as ET-AAS, F-AAS and GC-FID methods. The related weight of disease can be substantial, and interest in research on health effects and intervention in explicit populations and openness circumstances is significant for the development of control systems. Pollution control and determination is thusly a significant segment of disease control, and health experts and analytical chemistry specialists need to foster associations with different areas to recognize and carry out need interventions.

Acylserotonins – a new class of plant lipids with antioxidant activity and potential pharmacological applications

During analysis of components of baobab (Adansonia digitata) seed oil, several new fluorescent compounds were detected in HPLC chromatograms that were not found previously in any seed oils investigated so far. After preparative isolation of these compounds, structural analysis by NMR spectroscopy, UHPLC-HR-MS, GC-FID and spectroscopic methods were applied and allowed identification of these substances as series of N-acylserotonins containing saturated C22 to C26 fatty acids with minor contribution of C27 to C30 homologues. The main component was N-lignocerylserotonin and the content of odd carbon-atom-number fatty acids was unusually high among the homologues. The suggested structure of the investigated compounds was additionally confirmed by their chemical synthesis. Synthetic N-acylserotonins showed pronounced inhibition of membrane lipid peroxidation of liposomes prepared from chloroplast lipids, especially when the peroxidation was initiated by a water-soluble azo-initiator, AIPH. Comparative studies of the reaction rate constants of the N-acylserotonins and tocopherols with a stable radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) in solvents of different polarity revealed that N-acylserotonins showed similar activity to δ-tocopherol in this respect. The described compounds have been not reported before either in plants or in animals. This indicates that we have identified a new class of plant lipids with antioxidant properties that could have promising pharmacological activities.

Determination of Allyl Isothiocyanate in Mustard and Horseradish Extracts by Single Reference GC and HPLC Based on Relative Molar Sensitivities

The main component of the Mustard and Horseradish extracts, which are used as natural food additives in Japan, is allyl isothiocyanate (AITC). The determination of AITC using GC-FID is the official method employed in the quality control assessments for these products. In this method, a commercially available AITC reagent is used as a calibrant. However, 1H-quantitative NMR (qNMR) analysis revealed that the AITC reagents contain impurity. Therefore, we examined the GC-FID and HPLC-refractive index detector (LC-RID) method based on relative molar sensitivities (RMSs) to high-purity single reference (SR). The RMSs of AITC/SR under the GC-FID and LC-RID conditions were accurately determined using qNMR. The AITC in two types of food additives was quantified using qNMR, SR GC-FID, and SR LC-RID methods. Both SR GC-FID and SR LC-RID showed good agreement within 2% with the AITC content determined by direct qNMR.

High-Sensitivity Micro-Gas Chromatograph-Photoionization Detector for Trace Vapor Detection.

Rapid in situ detection and analysis of trace vapor concentrations at a sub-parts per billion to parts per trillion level remains a challenge for many applications such as indoor air-quality analysis and detection of explosives and narcotics. Micro-gas chromatography (μGC) together with a micro-photoionization detector (μPID) is a prominent method for portable analysis of complex vapor mixtures, but current μPID technology demonstrates poor detection performance compared to benchtop flame ionization detectors (FIDs). This work demonstrates the development of a significantly improved μPID with a sub-picogram detection limit (as low as ∼0.2 pg) comparable to or exceeding that of a benchtop FID, with a large linear dynamic range (>4 orders of magnitude) and robustness (high stability over 200 h of plasma activation). Based on this μPID, a complete μGC-PID system was built and tested on standard sample chromatograms in a laboratory setting to show the system's analytical capabilities and the detection limit down to sub-parts per trillion concentrations (as low as 0.14 ppt). Practical in-field chromatograms on breath and car exhaust were also generated to demonstrate applicability for in situ experimentation. This work shows that μGC-PID systems can be competitive with traditional GC-FID methods and thus opens a door to rapid trace vapor analysis in the field.

Differentiation between drug-type, fibre-type and intermediate-type in cannabis samples: HPLC-UV versus GC-FID

Cannabis sativa is known to be the most abused illegal drug worldwide. To date it is not only used as a medicine but has been established as a lifestyle product. The most relevant phytocannabinoids represent the ingredients delta-9-tetrahydrocannabinol (9-THC) and cannabidiol (CBD), whereby only 9-THC shows a psychoactive effect. Since 2017, the so-called CBD-hemp containing CBD as main ingredient is distributed in many countries as a legal alternative. In these products, 9-THC must not exceed a certain percentage. It is hardly possible to differentiate between THC-hemp and CBD-hemp presenting a major challenge for authorities. Therefore, there is the need to develop fast and efficient analysis methods to distinguish between fibre-type, drug-type and intermediate-type cannabis products. The aim of this study was to compare two simple and inexpensive HPLC-UV and GC-FID methods for their ability to quantify phytocannabinoids in dried cannabis plant material. For this purpose, a set of 37 fresh and dried cannabis samples randomly chosen from seizures of Austrian police was subject to complementary quantification of 9-THC and CBD. After having taken into account decomposition of certain phytocannabinoids, the result of this quantitative study showed good correlation between HPLC-UV and GC-FID regardless of quantifying cannabis leaves or buds.