Fish Extracts Research Articles

Penambahan Bubuk Ekstrak Ikan Haruan (Channa Striata) terhadap Kekuatan Tekan dan Perubahan Warna Glass Ionomer Cement (GIC) Tipe II Secara In Vitro

This study aims to prove the effect of adding haruan fish extract powder (Channa striata) on increasing the strength of type II Glass Ionomer Cement (GIC) filling materials. The method used is a laboratory experimental method with a post-test-only control group design approach. There are 6 treatment groups, namely the addition of 10mg, 20mg, 30mg, 40mg, 50mg, and without addition (pure). This research was conducted at the Dental Materials Laboratory, Faculty of Medicine, Dentistry Study Program, Mulawarman University and PT. Sucofindo Samarinda. The study was conducted from August 2022 to March 2023. The results showed that the greatest compressive strength of glass ionomer cement (GIC) was found in pure glass ionomer cement (GIC) without the addition of haruan fish extract which showed a compressive strength value of 37.0 MPa. The more amount of haruan fish extract powder added, the lower the strength of the glass ionomer cement will be. There is a difference in compressive strength between pure glass ionomer cement (GIC) without haruan fish extract powder and glass ionomer cement (GIC) with haruan fish extract. In conclusion, Glass Ionomer Cement (GIC) type II without the addition of fish extract powder must have the greatest compressive strength compared to the addition of 10 mg, 20 mg, 30 mg, 40 mg, and 50 mg. The addition of haruan fish extract powder decreased the strength of the glass ionomer cement and no color change occurred. 
 
 Keywords: Haruan Fish Extract Powder, Glass Ionomer Cement, Compressive Strength

Effect of pasak bumi (Eurycoma longifolia Jack), DHA, and seluang fish (Rasbora spp.) on neuroinflammation and neurotransmitter alterations in malnourished rats

Background: Malnutrition has detrimental effects on brain development, leading to neuroinflammatory and neurotransmitter disorders. A nutrient-rich diet is advocated to mitigate these effects. In South Kalimantan, natural resources such as pasak bumi (Eurycoma longifolia Jack.) and seluang fish (Rasbora spp.) are recognized for their potential antioxidant and anti-inflammatory properties. Objective: This study aimed to evaluate the effects of pasak bumi in comparison with seluang fish and DHA on neuroinflammation and neurotransmitter imbalances in malnourished rats. Methods: The study involved dividing malnourished rats into six experimental groups: (1) untreated control, (2) treated with pasak bumi extract, (3) treated with DHA, (4) treated with a combination of DHA and pasak bumi extract, (5) treated with seluang fish, (6) treated with a combination of seluang fish and pasak bumi extract; and a control group of normal rats receiving standard feed and placebo. The primary outcomes measured were levels of IL-6, TNF-α, and serotonin. Results: The study revealed that malnutrition in rats significantly elevated IL-6 and TNF-α levels. Treatments involving pasak bumi extract, both alone and in combination with DHA or seluang fish, reduced IL-6 levels. Similarly, combination of pasak bumi extract and DHA or seluang fish lowered TNF-α levels than single treatment of pasak bumi extract (p > 0.05). All treatments did not reduce the serotonin level. Conclusion: The findings of this study underscore the potent anti-inflammatory capabilities of pasak bumi extract, particularly when combined with DHA or seluang fish, in mitigating the inflammatory response in malnourished rats.

Tuna Ethanol Extract reduces CRP and Edema in Animal Models of Adjuvant Arthritis

Rheumatic arthritis is an inflammatory joint disease caused by an autoimmune process. Tuna contains many nutrients that act as antioxidants and anti-inflammatories in the form of vitamins, minerals, and essential fatty acids. This study aims to determine the effect of Tuna fish extract on inflammation in the legs of animal models of adjuvant arthritis. Twenty-four male Rattus norvegicus which were partitioned into P1 groups of no treatment rats, P2 groups of adjuvant arthritis (AA) rats, were nduced with Complete Freund's Adjuvant (CFA), P3 groups are rats with adjuvant arthritis who were given Thunnus obesus fish ethanol extricate orally for seven consecutive days from the primary day of the emergence of symptoms of AA, and given with the dosage of 12 grams/kg of body weight, beginning on the 22nd day. At the last day of the study, the CRP and swelling feet joints of all groups of rats were measured (on day 29). Tuna ethanol extracts significantly decrease the mean volume of edema (p = 0.034) and CRP levels (p=0.022) in adjuvant arthritis rats. The content of active ingredients in Tuna has been shown to inhibit the inflammatory process through the inhibition of cox-2, resulting in a reduction of the volume of animal leg joint edema and CRP levels in adjuvant arthritis trials. Administration of Tuna fish extract at a dose of 12g/kg BW can reduce leg edema of adjuvant arthritis experimental animals and serum CRP levels of adjuvant arthritis experimental animals.

Biodiesel production from by extraction of carp fish (Cyprinidae) oil and transesterification using CaO derived from limestone and eggshells as heterogeneous catalysts

The fish market produces a huge amount of fish waste and worsens the ecosystem’s condition when dumping into the environment while the treatment to create ecologically and economically acceptable biodiesel is one possible answer. So, this study evaluates the efficiency of three extraction techniques: Wet rendering, Soxhlet, and enzyme hydrolysis in extracting oil from carp fish. The highest yields of oil extraction were 38.61, 36.25, and 22.38% in the Soxhlet extraction, enzyme hydrolysis extraction, and wet rendering respectively. Raw fish oil’s physical and chemical characteristics were investigated, and GC-MS was used to determine its free fatty acid profile. This investigation also inquired about the preparation of calcium oxide (CaO) that is efficient in producing biodiesel from waste eggshells and limestone which were calcined at 950 °C for 2.5 hr, producing stable and high-purity CaO. XRD and FTIR were used to characterize the prepared catalyst while FESEM analysis of the catalyst’s surface structure and EDX spectra analysis of the catalyst’s elemental composition were both performed. The highest yields were 91.74 and 89.21% obtained using CaO derived from limestone and eggshells respectively. The optimum transesterification reaction conditions were a methanol to oil molar ratio of 9:1, 3.5 wt% CaO catalyst, 60 °C, and 90 min.

3-Epimers of Caribbean ciguatoxins in fish and algae

Ciguatera poisoning (CP) is endemic to several subtropical and tropical regions and is caused by the consumption of fish contaminated with ciguatoxins (CTXs). The recent discovery of Caribbean CTXs (C-CTXs) in Gambierdiscus spp. isolated from the Caribbean resulted in the identification of a precursor analogue, C-CTX5, that is reduced into C-CTX1. C-CTX5 has two reducible sites, a ketone at C-3 and hemiketal at C-56. Chemical reductions of C-CTX5 into C-CTX3/4 resulted in two peaks in the LC−HRMS chromatograms with a ratio that differed markedly from that observed in fish extracts and the reduction of C-CTX1 isolated from fish. Reduction of C-CTX5 should have produced four diastereoisomers of C-CTX3/4, prompting a more detailed study of the reduction products. LC–HRMS with a slow gradient was used to separate and detect the four stereoisomers of C-CTX3/4, and to determine the distribution of these analogues in naturally contaminated fish tissues and following chemical reduction of isolated analogues. The results showed that in naturally contaminated fish tissues C-CTX1/2 is a mixture of two diastereoisomers at C-3 and that C-CTX3/4 is a mixture of two pairs of diastereoisomers at C-3 and C-56. The data suggests that there is variability in the enzymatic reduction at C-3 and C-56 of C-CTXs in reef fish, leading to variations in the ratios of the four stereoisomers. Based on these findings, a naming convention for C-CTXs is proposed which aligns with that used for Pacific CTX congeners and will aid in the identification of the structure and stereochemistry of the different CTX analogues.

Snakehead Fish Extract in Increasing Skin Collagen & Fibroblasts

This study aim is to gain a deeper understanding of the mechanisms by which snakehead fish extract affects the skin, opening doors for the development of more effective and natural skincare products. The study employs a systematic literature review method using PRISMA strategy, identifying 156 relevant articles through platforms such as PubMed, Google Scholar, Scopus, SINTA, spanning from 2018 to 2023. Findings encompass increased VEGF production and NO levels with snakehead fish extract dosages, wounds healing potential of honey and snakehead fish skin combination in hydrogel formulations, increased fibroblast counts, reduced skin erythema, and decreased wound crust formation. Additionally, marine collagen exhibits promise in skin healing and bone repair. Collagen supplements, whether oral or topical, also prove beneficial in delaying skin aging. The study reveals crucial insights into the potential of snakehead fish extract in skincare and wound healing, supporting the development of more effective and natural skincare products.

Selective determination of 2-aminobenzothiazole in environmental water and organic extracts from fish and dust samples

In the present study, a homemade mixed-mode ion-exchange sorbent based on silica with embedded graphene microparticles is applied for the selective extraction of 2-aminobenzothiazole (NH2BT) followed by determination through liquid chromatography coupled to high-resolution mass spectrometry. The sorbent was evaluated for the solid-phase extraction of NH2BT from environmental water samples (river, effluent wastewater, and influent wastewater), and NH2BT was strongly retained through the selective cation-exchange interactions. Therefore, the inclusion of a clean-up step of 7 mL of methanol provided good selectivity for the extraction of NH2BT. The apparent recoveries obtained for environmental water samples ranged from 62 to 69% and the matrix effect from −1 to −14%. The sorbent was also evaluated in the clean-up step of the organic extract for the extraction of NH2BT from organic extracts of indoor dust samples (10 mL of ethyl acetate from pressurized liquid extraction) and fish (10 mL of acetonitrile from QuEChERS extraction). The organic extracts were acidified (adding a 0.1% of formic acid) to promote the cation-exchange interactions between the sorbent and the analyte. The apparent recoveries for fish samples ranged from 22 to 36% depending on the species. In the case of indoor dust samples, the recovery was 41%. It should be highlighted the low matrix effect encountered in such complex samples, with values ranging from −7 to 5% for fish and dust samples. Finally, various samples were analyzed. The concentration in river samples ranged from 31 to 136 ng/L; in effluent wastewater samples, from 55 to 191 ng/L; in influent wastewater samples, from 131 to 549 ng/L; in fish samples, from 14 to 57 ng/g dried weight; and in indoor dust samples, from <MQL to 114 ng/g.

An Effect of Giving Snakehead Fish Extract ( Channa striata ) on Albumin, Neutrophil, and Lymphocyte Levels in Hypoalbuminemia Patients

Background : Hypoalbuminemia is a condition where the albumin level in the blood falls below normal. Hypoalbuminemia is known as a marker of protein energy malnutrition, inflammation, infection, liver disease, kidney disease and postoperative stress. In Indonesia, malnutrition hospital data shows 40-50% of patients have hypoalbuminemia or are at risk of hypoalbuminemia, 12% of them are severe hypoalbuminemia, and the hospitalization period for patients with hospital malnutrition shows 90% longer than patients with good nutrition. Decreased albumin levels can be associated with a decrease in the immune system and slow healing of tissues/wounds due to infection. Neutrophils and lymphocytes whose numbers increase when inflammation occurs. Oral administration of high protein with snakehead fish extract ( Channa striata ) is an alternative treatment for hypoalbuminemia. Objective: The purpose of this study was to determine the effect of snakehead fish extract (Channa striata) on albumin, neutrophil, and lymphocyte levels in hypoalbuminemic patients. Method: Articles are compiled using a systematic method literature review , involving national and international journals by taking journals through Google scholar, Pubmed, Research Gate, Scient direct. Results: There was an effect of giving snakehead fish extract on albumin levels, neutrophil values, and lymphocyte values in hypoalbuinemic patients and experimental animals. Conclusion: The content of amino acids, albumin, Zn, Fe, Cu and unsaturated fatty acids in snakehead fish ( Ophiocephalus striatus) is quite effective as an effort to increase albumin levels, decrease neutrophil and lymphocyte values when inflammation occurs in hypoalbuminemic patients.&#x0D; &#x0D; Keywords: Channa striata extract , Albumin, Neutrophils, Lymphocytes, Hypoalbuminemia

Optimized protocol for the extraction of fish DNA from freshwater sediments

Optimized protocol for the extraction of fish <scp>DNA</scp> from freshwater sediments

Molecular mechanisms and potential applications of chondroitin sulphate in managing post-traumatic osteoarthritis.

Post-traumatic osteoarthritis (PTOA), a disorder of the synovium, subchondral bone, and cartilage that affects the entire joint, constitutes approximately 12% of all cases of symptomatic osteoarthritis. This review summarizes the pathogenetic mechanisms that underlie the positive influence of chondroitin sulphates (CSs) on PTOA as means of preventive and therapeutic treatment. Mechanisms of PTOA development involve chondrocytes undergoing various forms of cell death (apoptosis, pyroptosis, necroptosis, ferroptosis and/or necrosis). Chondroitin sulphates are a class of glycosaminoglycans that improve the structure and function of cartilage and subchondral bone, which is associated with their ability to decrease the activation of NF-κB and p38 MAPK, and up-regulate Nrf2. Standardized small fish extract (SSFE) is an example of the drugs that can attenuate NF-κB-mediated systemic inflammation, potentially helping to reduce joint inflammation and cartilage degradation, improve joint function, and alleviate pain and disability in patients with these conditions.

Pengukuran omega-3 pada ikan penja (Awaous sp.) asal Polewali Mandar Provinsi Sulawesi Barat

Omega-3 is an essential fatty acid necessary for human health. It has anti-inflammatory properties, reduces the risk of cardiovascular disease, and is needed for brain development. The three main omega-3 fatty acids are alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). Penja fish is a typical marine biota of Polewali Mandar, West Sulawesi, which is believed to be rich in omega-3. The research aimed to determine the omega-3 content in penja fish (Awaous sp.) from Polewali Mandar, West Sulawesi Province. Penja fish was extracted using the Soxhlet method, and then the fatty acid content was analyzed by GC-MS. The results of this study show that penja fish extract (Awaous sp.) contains omega-3 consisting of 2.64% linolenic acid, 0.94% eicosapentaenoic acid (EPA), and 1.17% docosahexaenoic acid (DHA). The total omega-3 content in penja fish (Awaous sp.) is 4.75%.

The effect of stabilizer type on the stability of snakehead fish (Channa striata) extract dispersion

Snakehead fish (Channa striata) extract in the form of dispersion is one of the innovations in the development of snakehead fish products. Snakehead fish extract dispersion is a product in the form of a mixture of syrup (dispersion) made from snakehead fish extract raw materials, honey, and other additives. However, the syrup/dispersion products produced in previous studies have not been completely stable. This is indicated by the presence of precipitation during dispersion storage. Therefore, it is necessary to choose the right type of stabilizer to overcome this. The aims of this study were to obtain the best stabilizer for producing a stable snakehead fish extract dispersion. The method in this study was to make a dispersion of snakehead fish extract with three different stabilizers, namely xanthan gum (0.3%), carrageenan (0.1%), and carboxymethyl cellulose (CMC) (0.6%). Then we analyzed the viscosity, phase separation ratio, and redispersibility. The results showed that xanthan gum has a better level of stability than carrageenan and CMC, with a viscosity value after storage of 1, 3, 5, and 7 days, respectively 53.97 mPa.s, 51.67 mPa.s, 51.50 mPa.s, and 51.17 mPa.s. While the viscosity value of carrageenan during storage is 1, 3, 5, 7 days, respectively 5.27 mPa.s, 5.20 mPa.s, 4.90 mPa.s, and 4.50 mPa.s, and storage of CMC 1, 3, 5, and 7 days, respectively 642.67 mPa.s, 505.33 mPa.s, 452.00 mPa.s, and 297.33 mPa.s. The results of the phase separation ratio analysis showed that the stabilizers xanthan gum and CMC did not experience phase separation during storage for 1, 3, 5, and 7 days. Meanwhile, the carrageenan stabilizer at storage 1, 3, 5, and 7 days has a successive phase separation ratio of 43.81%, 40.34%, 39.47%, and 39.47%. The results of the redispersibility analysis showed that the CMC stabilizer and xanthan gum were not redispersible during storage 1, 3, 5, and 7 days. While the carrageenan stabilizer has the same redispersibility value during storage 1, 3, 5, and 7 days is one time shuffling.

Chemical, Physical, and Hedonic Quality Characteristics of Fish Cereal from Toman Fish Albumin Extract (Channa micropeltes) as Functional Food

&lt;p&gt;&lt;em&gt;The nutritional value of fish cereal was determined by adding toman fish extract. Fish cereal is expected to be a useful food source rich in nutrients such as carbohydrates, vitamins, and minerals but rich in protein and albumin. Toman fish cereal is expected to be popular with people of all ages as a breakfast menu and can be a source of nutrition and increase endurance. The research aimed to obtain the formulation of toman fish albumin extract (Channa micropeltes) in fish cereal products as a functional food. This research was carried out in four stages: (1) preparation of fish albumin extract, (2) preparation of fish cereal; (3) Physical characteristics (hedonic test), fracture strength, and chemical characteristics (moisture content, ash content, protein content, fat content, and carbohydrate content), crude fibre content, albumin content, (4) determination of the best treatment using the Bayes method. The treatment with the addition of toman fish albumin consisted of 5 treatments with two replications. The treatment was carried out namely A1 0%, A2 30%, A3 35%, A4 40%, and A5 45%. The results showed that the highest protein content was around 4.57–13.33% and the albumin content of fish cereal was around 4.86–6.61%. This showed that fish cereal can be a functional food in increasing the immune system. Based on the Bayes test, the selected toman fish cereal formula was formula A2, namely 30% albumin extract.&lt;/em&gt;&lt;/p&gt;

Non-inferiority trial of Channa striata extract on endothelial glycocalyx layer protection in septic patients

Background: Sepsis causes significant damage to the endothelial glycocalyx layer, which albumin can protect against. Unfortunately, human albumin is quite expensive, so cheaper substitutes must be found without sacrificing effectiveness. Albumin extract called snakehead fish extract (SHFE) is one that is commonly used in Indonesia It is also called Channa striata extract. We conducted a non-inferiority study between human albumin and SHFE in protecting the glycocalyx layer through syndecan-1 levels.&#x0D; Methodology: It was a prospective cohort study of adult patients with early sepsis, treated at ICU of Dr Moewardi Hospital, between May and October 2021. The study group was divided by giving Channa striata extract 15 mg/day for two days and the control group was given 20 grams of human albumin on the other day, and then syndecan-1 levels were compared before treatment and on the third day of the study.&#x0D; Results: We discovered a statistically significant decrease in syndecan-1 levels in both the study (P = 0.013) and control groups (P = 0.027) from the 44 samples we obtained.&#x0D; Conclusion: In both groups, there was a significant decrease in syndecan-1 levels, proving that Channa striata extract is not inferior to human albumin in terms of glycocalyx layer protection. And it has a higher economic value because the price is lower.&#x0D; Key words: Sepsis; Channa striata; Albumin; Syndecan-1; Glycocalyx&#x0D; Citation: Permana SA, Hartono H, Purwanto B, Indarto D. Non-inferiority trial of channa striata extract on endothelial glycocalyx layer protection in septic patients. Anaesth. pain intensive care 2023;27(4):523−527; DOI: 10.35975/apic.v27i4.2278&#x0D; Received: May 15, 2023; Reviewed: June 22, 2023; Accepted: June 22, 2023

Hydrogel-based Bioelectronic Tongue for the Evaluation of Umami Taste in Fermented Fish.

Bioelectronic tongues based on umami taste receptors have recently been reported for versatile applications such as food analyses. However, their practical applications are still limited, partly due to their limited stability and non-specific responses in real sample environments. Herein, we have developed a hydrogel-based bioelectronic tongue for the sensitive assessment of umami intensity in fish extract samples. In this study, the T1R1 venus flytrap of an umami taste receptor was immobilized on the gold floating electrodes of a carbon nanotube-based field-effect transistor. A polyacrylamide conducting hydrogel film was further hybridized on the sensor surface via physical adsorption, which could provide a good physiological environment to maintain the activity of receptors due to its excellent hydrophilicity and biocompatibility. The bioelectronic tongue with a receptor-embedded hydrogel structure showed a sensitive detection of umami substances down to 1 fM, and it also had a wide detection range of 10-15-10-2 M for monosodium glutamate and disodium inosinate, which covers the human taste threshold. More importantly, the proposed sensor could significantly reduce the non-specific binding of non-target molecules to a carbon nanotube channel as well as exhibit long-term stability, enabling sensitive detection of umami substances even in fish extract samples. Our hydrogel-based bioelectronic tongue provides a promising platform for future applications such as the flavor evaluation of foods and beverages.

Toxic or beneficial? What is the role of food‐relevant selenium species selenoneine?

Toxic or beneficial? What is the role of food‐relevant selenium species selenoneine?

Documentation of phytochemicals, bio-toxicity and haemato-physiological responses at sub-lethal concentrations of Parthenium hysterophorus hydroethanolic extract in Cyprinus carpio: a concept of waste valorization as a phyto-ichthyotoxin.

In the present study, phytoextraction of a weed plant, Parthenium hysterophorus, was performed through aqueous, alcoholic and hydroethanolic (80%) solvents followed by phytochemical profiling and evaluation of median lethal concentration (LC50) of hydroethanolic extract in a freshwater fish, common carp (Cyprinus carpio). Haemato-physiological response was also evaluated based on LC50 (18.99mg L-1) at two sub-lethal concentrations of extract [T1: 0.379mg L-1 (LC50/50), T2: 0.759mg L-1 (LC50/25) and a control: devoid of extract] at three intervals (24, 48 and 96h). The study revealed toxic constituents in extracts and the superior extraction ability of hydroethanolic solvent which was chosen for further biological characterisation, particularly on haematotoxicity. The anti-bacterial assay revealed the inhibitory capacity of the extract, whereas the phyto-haemagglutination assay, haemagglutination limit test and haemolytic activity revealed clumping, agglutination (at 1/96th dilution) and lytic capability of extract, respectively. Later, in vivo analyses revealeda significant modulation in haemato-immunological and serum biochemical parameters upon hydroethanolic extract exposure. In conclusion, the present study emphasises locally available gajar ghas, P. hysterophorus as a non-chemical phyto-ichthyotoxin towards sustainable aquaculture.

WATER PHASE CONCENTRATE SOLUBILITY TEST SNAKEHEAD FISH EXTRACT (Channa Striata) IN ORGANIC COSOLVENT

Snakehead fish extract has been recognized for its potential in healing postoperative wounds and burns. The extract contains albumin, which aids in tissue formation and healing processes. To create a high-quality wound healing drug, the extract needs to be formulated into a concentrated form and combined with a solvent that can effectively dissolve its active substances. Understanding the solubility profile of the extract and the composition of the solvent is crucial for this purpose. This study aims to determine the solubility profile of the aqueous phase concentrate of snakehead fish extract in an organic co-solvent. Solubility is an important parameter as it can impact the release rate and therapeutic effectiveness of the drug. A solubility test was conducted to assess the solubility of the aqueous phase concentrate in various solvents. The results of the solubility test indicated that the aqueous phase concentrate of snakehead fish extract exhibited good solubility in aquadest (20%), slight solubility in propylene glycol (8.5%) and honey (2%), and was insoluble in ethanol, glycerin, mannitol, and sorbitol (0%). These findings provide valuable information for formulating the extract into a wound healing drug with an appropriate solvent composition, ensuring optimal solubility and therapeutic efficacy according to pharmacopeial standards. Keywords: Concentrate; Aqueous phase of snakehead fish extract; Solubility; Solvent; Polarity Properties

Effect of Organic and Liquid Manures on Productivity and Profitability of Blackgram

Background: Blackgram being important crop among pulse is gaining popularity and thus requiring suitable crop production practices especially under organic farming. In this context, this study was proposed with the objective to investigate the optimal nutrient management practices in organic blackgram by utilizing various organic nutrient sources. Methods: Field experiments were conducted at two different locations in spilt plot design. The treatments consisted of 4 levels of organic manure viz., Farm Yard Manure (FYM) @ 12.5 t ha-1, Vermicompost (VC) @ 2.5 t ha-1, Enriched FYM (EFYM) @ 750 kg ha-1 and Enriched VC (EVC) @ 1 t ha-1 as main plot treatments and five levels of foliar spray of liquid manure viz., two foliar sprays of Panchagavya @ 3% and Fermented egg extract (FEE), Fermented fish extract (FFE), Farmers effective microorganisms (FEM), Jeevamruth each @ 5%. Result: Results revealed that application of EFYM and VC, produced higher yield parameters and grain yield, respectively under main plot organic manure addition at site-I and II. Among the sub plots liquid manure treatments Panchagavya spray has produced higher grain yield along with the yield parameters at both the sites. Among the organic manures higher economic returns were obtained with the application of EFYM at both the sites while among the liquid manures better returns were obtained with the application of Panchagavya at site-I and Panchagavya or FEM spray at site-II.

A simple and sensitive method for the determination of methylene blue and its analogues in fish muscle using UPLC-MS/MS

A new, simple and sensitive method for determining and confirming methylene blue and its analogues such as azure A, azure B, azure C, thionine, and new methylene blue in fish muscles have been developed. The method is based on acetonitrile extraction followed by extract purification using dispersive solid-phase extraction (dSPE) with basic aluminium oxide (ALN) and solid-phase extraction (SPE) using primary and secondary amines (PSA) sorbent in matrix adsorption mode. The separation and detection of the dyes in the fish extract are achieved within 5 min by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) using an octadecyl analytical column with a mixture of acetonitrile, methanol and 0.1% formic acid as a mobile phase in gradient elution. The developed method has been in-house validated according to European law. The method recovery for fish muscle was 98.3–103.1%, whereas the decision limit (CCα) was from 0.45 to 0.49 µg kg−1.