Commercial Preparations Research Articles

Bowman-Birk inhibitor in soybean: Genetic variability in relation to total trypsin inhibitor activity and elimination of Kunitz trypsin inhibitor

Bowman-Birk inhibitor (BBI) is a soybean seed serine protease inhibitor whose nutraceutical properties far exceed its anti-nutritional property arising from its trypsin and chymotrypsin inhibitor activity. High BBI soybean genotypes are sought for the commercial preparation of this nutraceutical biomolecule while low BBI content in Kunitz trypsin inhibitor (KTI)-free genetic background is desirable in soymeal manufacturing. In the present investigation, monoclonal antibody assay of 95 soybean genotypes revealed 11-fold genetic variation for BBI concentration. The study led to the identification of 3 very high (>20 mg/g defatted soy flour) and 5 very low BBI (< 4 mg/g defatted soy flour) soybean genotypes. Contribution of BBI to total trypsin inhibition activity ranged from 2.2 to 53.5% with average value of 11.6%. Genotypes with varying level of BBI raised consecutively for two years exhibited non-significant (p>0.05) effect of growing year on the trait. BBI content in BC3F2 derived KTI-free lines was at par with the recurrent parent. Low and high BBI content genotypes with diverse genetic background identified in the study may be exploited to develop mapping population to identify genomic regions underlying BBI in soybean.

Are multiplexed metabarcoding panels comparable to individual marker gene library preparations?

Analysis of multiple marker genes using metabarcoding of environmental DNA (eDNA) can offer information greater than that from sequencing single marker genes, such as responses from across the phylogenetic tree to environmental gradients (Cordier et al. 2019). Furthermore, multiple regions of the same gene can be sequenced to improve phylogenetic resolution (Fuks et al. 2018). However, separate amplification reactions and library preparation steps for each marker can be costly and time consuming. Here, we have designed and optimised a multiplex panel of four marker genes (two regions of 18S rRNA gene, one region of the 16S rRNA gene and one region of the rbcL gene). By combining steps into a single reaction, the labwork required is decreased, reducing cost and time. This multiplex is compared with a widely available commercial microbial (bacterial and fungal) screening panel and individual library preparations of each marker gene.

Combining culinary art with food science

Combining culinary art with food science

A supercritical fluid workflow for the quality assessment of herbal drugs and commercial preparations from Rhodiola rosea.

IntroductionPreparations from the Rhodiola rosea are experiencing an increase in popularity: extracts of dried roots and rhizomes are used as adaptogen to treat stress, fatigue, and weakness. To meet high pharmaceutical standards, fast and reliable methods to assess phytochemical variations in respect of quality control are needed.ObjectiveThe aim of this study was to extract and quantify seven characteristic secondary metabolites of R. rosea , namely p‐tyrosol (1), rosin (2), rosiridin (3), salidroside (4), rosarin (5), rosavin (6), and tricin‐5‐O‐β‐d‐glucopyranoside (7) in 24 herbal drugs and seven commercial preparations using a newly established supercritical fluid workflow.MethodsThe developed protocol allowed for an exhaustive extraction of compounds 1–7 using 60% carbon dioxide (CO2) and 40% methanol. The constituents were analysed on an ultra‐high‐performance supercritical fluid chromatography (UHPSFC) instrument using a charged surface hybrid fluoro‐phenyl (CSH FP) column (3.0 mm × 100 mm, 1.7 μm; mobile phase: CO2 and methanol).ResultsThe seven compounds were separated in a remarkably short time (< 3.5 minutes). For their quantitation, good results in terms of selectivity, linearity (R 2 ≥ 0.99), precision (intraday ≤ 3.03%, interday ≤ 5.17%) and accuracy (recovery rates 96.6–102.4%) were achieved using selected ion recording on a Quadrupole Dalton (QDa) mass detector.ConclusionThe quantitative analysis of the investigated herbal drugs showed a highly differing metabolite pattern which was also observed in the investigated commercial products. None of the commercial dietary products met the declared content of rosavins and salidroside. The developed and validated protocol offers a novel and reliable method to assess the quantitative composition of Rhodiola herbal drugs and preparations.

Yellow Lake Pigments from Weld in Art: Investigating the Winsor &amp; Newton 19th Century Archive

Weld (Reseda luteola) was one of the main sources of yellow dyes used for dyeing textiles and to prepare artists’ pigments in Europe until the 19th century. For the first time, this work explores the technology of preparing weld lake pigments in the 19th century by Winsor &amp; Newton (W&amp;N), a renowned supplier of artists’ materials. Five recipes were discovered in the W&amp;N 19th century Archive Database and reconstructed in the laboratory. W&amp;N was extracting weld in neutral and basic media, and preparing the insoluble lake by complexation with Al3+ in the form of alum (KAl(SO4)2•12H2O) or hydrated alumina (Al(OH)3). Five yellow lake pigments were successfully obtained and characterized by High-Performance Liquid Chromatography-Diode Array Detector (HPLC-DAD) and Fourier Transform Infrared Spectroscopy (FTIR). Their chromatographic profiles display as main yellows, luteolin 7-O-glucoside (Lut-7-O-glu) or both Lut-7-O-glu plus luteolin 3′,7-O-glucoside (Lut-3’,7-O-glu). In two of the processes, the presence of gypsum (CaSO4•2H2O) was unequivocally detected by FTIR, being formed as a by-product. This work offers the first identification of weld lake pigments’ characteristic infrared bands. The W&amp;N Database proved again to be a unique source of information on 19th-century artists’ materials and their commercial preparation. The knowledge gain is essential to ensure effective conservation and authentication procedures.

Development of a Methodology for Estimating the Ergosterol in Meat Product-Borne Toxigenic Moulds to Evaluate Antifungal Agents.

Antifungal agents are commonly used in the meat industry to prevent the growth of unwanted moulds, such as toxigenic ones, on dry-cured meat products. For enhancing the application of antifungals, their mode of action must be evaluated. Their effect on the mould ergosterol content is one of the most studied ones, since it is the target site of some commercialised antifungals or of those that are in development. The aim of this study was to develop a methodology for determining how the antifungal agents used in the meat industry work. A method for analysing ergosterol was firstly developed using high-performance liquid chromatography with fluorescence detection coupled to a diode array detector (HPLC-FLD/DAD). The chromatographically optimised conditions (gradient and mobile phases) allowed us to reduce the time per analysis with respect to previously published methods up to 22 min. Withing the six checked extraction methods, method 5, showing the best mean recovery values (99.51%), the shortest retention time (15.8 min), and the lowest standard deviation values (9.92) and working temperature (60 °C), was selected. The limit of detection and limit of quantification were 0.03 and 0.1 µg/mL, respectively. All the validation parameters corroborated the method’s suitability. Finally, its feasibility for evaluating the effect of a commercial antifungal preparation (AP) and different herbs that are frequently added to meat products on the ergosterol content of several toxigenic moulds was studied. Differences at the strain level were obtained in the presence of AP. Moreover, the addition of herbs significantly reduced the ergosterol content in Penicillium nordicum up to 83.91%. The developed methodology is thus suitable for screening the antifungals’ role in altering mould ergosterol biosynthesis before their application in real meat products.

Anti-SARS-CoV-2 Antibodies Within IVIg Preparations: Cross-Reactivities With Seasonal Coronaviruses, Natural Autoimmunity, and Therapeutic Implications

Introduction: Cross-reactivity to SARS-CoV-2 antigenic peptides has been detected on T-cells from pre-pandemic donors due to recognition of conserved protein fragments within members of the coronavirus's family. Further, preexisting antibodies recognizing SARS-CoV-2 with conserved epitopes in the spike region have been now seen in uninfected individuals. High-dose Intravenous Immunoglobulin (IVIg), derived from thousands of healthy donors, contains natural IgG antibodies against various antigens which can be detected both within the IVIg preparations and in the serum of IVIg-receiving patients. Whether IVIg preparations from pre-pandemic donors also contain antibodies against pre-pandemic coronaviruses or autoreactive antibodies that cross-react with SARS-CoV-2 antigenic epitopes, is unknown.Methods: 13 samples from 5 commercial IVIg preparations from pre-pandemic donors (HyQvia (Baxalta Innovations GmbH); Privigen (CSL Behring); Intratect (Biotest AG); IgVena (Kedrion S.p.A); and Flebogamma (Grifols S.A.) were blindly screened using a semi-quantitative FDA-approved and validated enzyme-linked immunosorbent assay (ELISA) (Euroimmun, Lubeck, Germany).Results: Nine of thirteen preparations (69.2%), all from two different manufactures, were antibody-positive based on the defined cut-off positivity (index of sample OD to calibrator OD > 1.1). From one manufacturer, 7/7 lots (100%) and from another 2/3 lots (67%), tested positive for cross-reacting antibodies. 7/9 of the positive preparations (77%) had titers as seen in asymptomatically infected individuals or recent COVID19-recovered patients, while 2/9 (23%) had higher titers, comparable to those seen in patients with active symptomatic COVID-19 infection (index > 2.2).Conclusion: Pre-pandemic IVIg donors have either natural autoantibodies or pre-pandemic cross-reactive antibodies against antigenic protein fragments conserved among the “common cold” - related coronaviruses. The findings are important in: (a) assessing true anti-SARS-CoV-2-IgG seroprevalence avoiding false positivity in IVIg-receiving patients; (b) exploring potential protective benefits in patients with immune-mediated conditions and immunodeficiencies receiving acute or chronic maintenance IVIg therapy, and (c) validating data from a recent controlled study that showed significantly lower in-hospital mortality in the IVIg- treated group.

Green spectrofluorimetric determination of salmeterol xinafoate in its pure forms, medicinal commercial formula, and human plasma: Application for stability studies.

A natural, accurate, extremely rapid, and precise spectrofluorometric method has been developed and validated for determination of salmeterol (SAL) xinafoate in its medicinal commercial form and spiked human plasma. The native SAL fluorescence has been measured at 415 nm (after 340 nm as excitation) in distilled water. Different factors affecting the native fluorescence consistency of SAL were surveyed and optimized. The suggested procedure was capable of SAL determination over concentrations ranging 200-2000 ng.mL-1 with excellent correlation coefficient of 0.9995. The limits of detection and quantification were estimated as 44.44 ng mL-1 and 134.66 ng mL-1 , respectively. The method has good accuracy and precision. The green spectrofluorometric method was used for determination of SAL in its commercial preparations and the results were in accordance with other reported methods regarding accuracy and precision. Moreover, the proposed procedure was enforced for stability indicating assay of SAL and for SAL determination in spiked human plasma. Nearly no cost, high sensitivity, and wide application make the proposed method ideally suited for analysis of SAL in quality control laboratories.

Commercial preparations of pesticides exert higher toxicity and cause changes at subcellular level in earthworm Eisenia andrei

BackgroundThe environmental impact of pesticides has been an increasingly discussed issue over the last decades. Constant usage of pesticides presents a burden for soil and causes a decrease in its health, including the negative effects on earthworms which are indicators for soil quality. The objective of this research was the assessment of the effects of two insecticides and two herbicides on the earthworm Eisenia andrei. Namely, the following active ingredients and respective commercial preparations were investigated: esfenvalerate (Sumialfa), thiacloprid (Calypso), dimethenamid-p (Frontier) and prosulfocarb (Filon). Lethal concentrations (48 h) of both active ingredient and commercial preparations were determined using the filter paper contact test.ResultsThe results showed that Calypso and Frontier were significantly more toxic than the active ingredient. Therefore, all further measurements were performed after exposure of earthworms to the commercial preparations of the pesticides. Specifically, several enzymatic biomarkers and multixenobiotic resistance activity were assessed. Additionally, a fluorescence-based assay for the determination of oxidative stress was established. Significant changes were detected for catalase, carboxylesterase and multixenobiotic activities after 48-h exposures. Also, a significant change in oxidative stress parameters could be observed for both Calypso and Frontier.ConclusionsThe obtained results show that commercial preparations can be more toxic than the active ingredients, and the formulations being distributed in the environment can affect earthworms on a molecular level already after short exposures. This emphasizes the importance of a more integrated eco-toxicological assessment of commercial pesticide preparations not to underestimate their effects on the environment.

Spectroscopic characteristics of colloidal solutions of metal nanoparticles

Nanoparticles are finding more and more practical applications in various fields of human activity, including veterinary and medicine. Due to the fact that the effectiveness of activity of colloidal solutions of nanoparticles is directly related to the state of aggregation of nanosized particles, it is urgent to use fast and convenient methods for assessing the physicochemical characteristics of such preparations. Nanoparticles have unique optical properties that depend on their size and shape. They can be determined by the refractive index of the light on the surface of the nanoparticles in a phenomenon known as plasmon resonance, which makes UV-Vis spectroscopy a valuable tool for studying and evaluating the characteristics of nanomaterials. Goal of the study is to study the optical characteristics of several samples of colloids of nanoparticles of noble metals (silver) and bioelements (copper, silicon dioxide) in order to determine the possibility of further application of UV-Vis spectroscopy for evaluation of activity and stability of colloidal solutions of nanoparticles. Commercial preparations based on nanoparticles of noble metals (silver) or bioelements (copper, silicon dioxide) in working dilutions recommended by manufacturers were used to study optical characteristics of the colloidal solutions. Optical density and absorption spectra were determined at the wavelengths (nm): 300-800 nm. The f plasmon surface resonance has been found in all test preparations, while all of them exhibited obvious nonlinear optical properties. The most pronounced plasma resonance peak is found in the colloidal solution of silver nanoparticles within a wavelength of 420 nm. In the case of a colloidal solution of copper nanoparticles, the peak of plasmon resonance was less pronounced and had a red shift (peak at 560 nm). In the colloidal solution of silicon silica, the plasmon resonance was less pronounced than other test preparations, being shifted to the blue side of the spectrum (360 nm).

Effect of Simulated Gastrointestinal Tract Conditions on Survivability of Probiotic Bacteria Present in Commercial Preparations.

Probiotics are recommended, among others, in the diet of children who are under antibiotic therapy, or that suffer from food allergies or travel diarrhea, etc. In the case of toddlers taking probiotic preparations, it is highly recommended to first remove the special capsule, which normally protects probiotic strains against hard conditions in the gastrointestinal tract. Otherwise, the toddler may choke. This removal can impair probiotic survival and reduce its efficacy in a toddler’s organism. The aim of this study was to evaluate the survivability of five strains of lactic acid bacteria from the commercial probiotics available on the Polish market under simulated conditions of the gastrointestinal tract. Five probiotics (each including one of these strains: Bifidobacterium BB-12, Lactobacillus (Lb.) rhamnosus GG, Lb. casei, Lb. acidophilus, Lb. plantarum) were protective capsule deprived, added in a food matrix (chicken–vegetable soup) and subjected under simulated conditions of the gastric and gastrointestinal passage. Strain survivability and possibility to growth were evaluated. Obtained results showed that, among all analyzed commercial probiotic strains, the Lb. plantarum was the most resistant to the applied conditions of the culture medium. They showed a noticeable growth under both in vitro gastric conditions at pH 4.0 and 5.0, as well as in vitro intestinal conditions at all tested concentrations of bile salts.

Comprehensive analysis of lactoferrin N-glycans with site-specificity from bovine colostrum using specific proteases and RP-UHPLC-MS/MS

Bovine Lf (bLf) possesses five N-glycosylation sites having diverse glycans that alter over lactation and are extensively characterised in milk. However, detailed site-specific characterisation in colostrum is limited. In the present study, the glycan diversity in colostrum bLf from native and commercial preparations was determined by generating N-glycopeptides using trypsin and a combination of trypsin/chymotrypsin followed by RP-UHPLC-Q-TOF MS/MS. The glycan diversity at N300KS site is poorly understood and the analysis revealed the presence of mostly biantennary complex glycans with core fucosylation and differential sialylation. N252NS and N564DT sites were found to possess predominantly high mannose glycans. A range of complex, hybrid and high mannose glycans were determined at N387VT and N495QT sites. Glycan heterogeneity with relative abundance was determined on all glycosylation sites in bLf. In addition, the glycan diversity of lipocalin-2 (N85VT) that might have co-purified in the commercial colostrum bLf sample was also elucidated.

Phage phiKZ-The First of Giants.

The paper covers the history of the discovery and description of phiKZ, the first known giant bacteriophage active on Pseudomonas aeruginosa. It also describes its unique features, especially the characteristic manner of DNA packing in the head around a cylinder-shaped structure (“inner body”), which probably governs an ordered and tight packaging of the phage genome. Important properties of phiKZ-like phages include a wide range of lytic activity and the blue opalescence of their negative colonies, and provide a background for the search and discovery of new P. aeruginosa giant phages. The importance of the phiKZ species and of other giant phage species in practical phage therapy is noted given their broad use in commercial phage preparations.

A New Application of PARAFAC Model to UPLC Dataset for the Quantitative Resolution of a Tri-Component Drug Mixture.

In the presented work, a three-way analysis of ultra-performance liquid chromatography-photodiode array (UPLC-PDA) dataset was performed by parallel factor analysis (PARAFAC) for quantitatively resolving a ternary mixture containing paracetamol and methocarbamol with indapamide selected as an internal standard in their co-eluted chromatographic conditions. Paracetamol and methocarbamol were quantified in the working range between 3-24 and 5-50μg/mL by applying PARAFAC decomposition to UPLC-PDA data array obtained under unresolved chromatographic peak conditions. To compare the experimental results provided by co-eluted UPLC-PARAFAC method, an ordinary UPLC method was developed ensuring proper separation of the peaks. The performance of both PARAFAC and ordinary UPLC methods were assessed by quantifying independent test samples, intra- and inter-day samples and spiked samples of pharmaceutical preparations. Then, both methods were applied for quantitative estimation of the related drugs in a commercial pharmaceutical preparation. In this study, PARAFAC method was proved to be a very powerful alternative for the quality control of pharmaceutical preparations containing paracetamol and methocarbamol even in their co-eluted chromatograms with high precision and accuracy in a short chromatographic runtime of 1.2min.

Effect of container shape on freeze concentration of apple juice.

Concentrating fruit juices by freezing supports the maintenance of both nutrients and flavors. Development of the freezing concentration process has introduced equipment such as centrifuge or block freezing systems, which are suitable for large-scale commercial processing. However, the necessary characteristics of freeze concentration methods for juices include simplicity and low cost. This study examined the effects of different container shapes on the processes of freezing and melting. The shape of the container was found to be more important than the melting temperature, across a relatively large scale. Furthermore, the nutrient procyanidin B2 and saccharides were concentrated. The methods concentrated juice components under low cost conditions without complex equipment. This research thus not only offers benefits for commercial juice preparation but also provides new insight into effects of shape differences in concentration technologies.

Effects of ultrasound on the enzymatic degradation of pectin

Ultrasound-assisted enzymatic maceration (UAEM) has gained considerable interest in the fruit juice industry, owing to its potential to increase juice yield and content of polyphenols while simultaneously saving time and energy. In this study, the effects of UAEM (ultrasonic probe, 20 kHz, 21 W*cm−2 and 33 W*cm−2) on pectin degradation in a continuous circulation system were investigated over 60 and 90 min. Main pectinolytic enzymes activities of (polygalacturonase, pectin lyase and pectin methylesterase) of a commercial enzyme preparation were examined for individual synergistic effects with US. Pectin hydrolysis by UAEM differed significantly compared to treatment with ultrasound or enzymes alone regarding the profile of degradation products compared to treatment with ultrasound or enzymes alone. Ultrasound fragmented pectin to less branched oligomers of medium molecular weight (Mp approx. 150 kDa), which were further degraded by pectinolytic activities. The low molecular weight fraction (<30 kDa), which is known to be beneficial for juice-quality by adding nutritional value and stabilizing polyphenols, was enriched in small oligomers of homogalacturonan-derived, rhamnogalacturonan I-derived, and rhamnogalacturonan II-derived residues. Synergistic effects of ultrasound application enhanced the effective activities of polygalacturonase and pectin lyase and even prolonged their performance over 90 min, whereas the effective activity of pectin methylesterase was not affected. Final marker concentrations determined by each enzyme assay revealed a considerable higher total process output after UAEM treatment at reduced temperature (30 °C) comparable to the output after conventional batch maceration at 50 °C. The obtained results demonstrate the high potential of UAEM to produce high-quality juice by controlling pectin degradation while reducing process temperature and equally highlight the matrix and enzyme specific effects of a simultaneous US treatment.

Use of Endoglucanase and Accessory Enzymes to Facilitate Mechanical Pulp Nanofibrillation

Although selective enzyme treatments have been used to successfully fibrillate chemical pulps, high lignin-containing mechanical pulps have proven to be more recalcitrant. When a bleached chemi-thermomechanical pulp (BCTMP) was sulfonated prior to enzymatic treatment, relatively good fibrillation was achieved, although some pulp hydrolysis occurred after 6 h hydrolysis when using a commercial cellulase enzyme preparation (Cellic CTec 3). To try to minimize pulp losses, various enzyme cocktails, including endoglucanase (EG), xylanase, mannanase, and lytic polysaccharide monooxygenase (LPMO), were assessed for their ability to enhance fibrillation while minimizing cellulose hydrolysis. It was apparent that the yield as well as the zeta potential of the lignin-containing cellulose nanofibrils increased with enzyme treatment. This was likely due to an increase in surface charge and a decrease in particle size after LPMO and hemicellulase treatments, respectively. When carbohydrate-binding modules (CBMs) were used to quantify fiber changes, it was apparent that sulfonation had increased the accessibility of enzymes, while the combined action of the hemicellulases and LPMO increased EG accessibility to the less-ordered regions of the mechanical pulp, resulting in enhanced fibrillation. This work described, for the first time, the synergistic action of EG and various accessory enzymes enhancing mechanical pulp nanofibrillation.

ВИДІЛЕННЯ ІНГІБІТОРА ТРИПСИНУ НАСІННЯ ЛЮЦЕРНИ З ДОПОМОГОЮ АФФІННОЇ ХРОМАТОГРАФІЇ

A protein inhibitor of trypsin of the cultivar Eva alfalfa seeds was isolated and characterized. It has significant antiproteolytic activity and is promising for the creation of compositions that are designed to correct nutrition in various conditions, accompanied by increased activation of proteolytic enzymes. The stages of purification of the trypsin inhibitor from alfalfa seeds included: extraction of 0.05 M borate buffer (pH 7.6), fractionation of the protein component of the extract with ammonium sulfate, followed by dialysis of the fraction between 75% and 100% saturation and affinity chromatography on a biospecific sorbent trypsin - sepharose 4B. Calculations showed that from 100 g of alfalfa seeds of the cultivar Eva, 67.14 mg of trypsin inhibitor was obtained, the inhibitory activity of which is 27.6 IU / mg protein. Experimental data show that the obtained inhibitor reduces trypsin activity by 63.05% with a weight ratio of inhibitor: enzyme of 1: 1. It is known from literature that a soybean trypsin inhibitor reduces trypsin activity by 30%. The isolated trypsin inhibitor has significant inhibitory activity, the value of which is higher than the inhibitory activity of the plant trypsin inhibitor from soybeans. Affinity chromatography allowed us to obtain a pure, homogeneous trypsin inhibitor, which has significant antiproteolytic activity, which can be compared with the antiproteolytic activity of known plant trypsin inhibitors. From the obtained experimental data, it follows that both, the obtained inhibitor and the inhibitor, which is part of a commercial preparation, effectively reduce trypsin activity, slightly inhibit chymotrypsin activity, and do not affect α-amylase and protease activity. The above data indicate that the trypsin inhibitor of alfalfa seeds of the cultivar Eva is not a bifunctional inhibitor, since it does not affect the activity of amilolytic enzymes. The isolated inhibitor belongs to proteins with a high degree of hydrophobicity. The composition of amino acids, the side chains of which can take part in the formation of hydrogen bonds, is 25% of the total number of amino acid residues of the alfalfa seed trypsin inhibitor. For STI, this value is 23%.

Dietary exposure to trace elements (B, Ba, Li, Ni, Sr, and V) and toxic metals (Al, Cd, and Pb) from the consumption of commercial preparations of Spirulina platensis.

Spirulina is a multicellular cyanobacterium that is consumed as a dietary supplement. The content of trace elements (B, Ba, Li, Ni, Sr, V) and toxic metals (Al, Cd, Pb) was determined in 24 spirulina samples marketed in two different formulations (tablets and powder) by ICP-OES (inductively coupled plasma optical emission spectrometry). The highest element concentration was found in the powder presentation, except for Li. The powder presentation element levels (mg/kg dry weight) were Al (28.1), Sr (10.3), B (1.73), Li (1.47), Ba (1.25), Ni (0.63), Pb (88.1 μg/kg dw), Cd (37.2 μg/kg dw), and V (22.9 μg/kg dw). Considering an adult with a body weight of 68.48 kg and the posology guidelines (14 tablespoons per week), the consumption of powdered spirulina contributes greatly to the Al intake by 2.04% of its TWI (tolerable weekly intake) set at 1 mg/kg body weight/week, followed by Cd with 1.05% of its TWI set at 2.5 μg/kg bw/week. Pb intake represents 1.05% of the BMDL (benchmark dose) level associated with nephrotoxicity and 0.44% of the BMDL associated with cardiovascular effects. This assessment suggests that spirulina consumption does not pose risks to the consumer as far as exposure to toxic metals (Al, Cd, Pb) is concerned. However, the presence of trace elements and toxic metals in spirulina preparations should be monitored to ensure its quality and safety.

Box-Behnken Assisted Validation and Optimization of an RP-HPLC Method for Simultaneous Determination of Domperidone and Lansoprazole

A highly specific, accurate, and simple RP-HPLC technique was developed for the real-time quantification of domperidone (DOMP) and lansoprazole (LANS) in commercial formulations. Chromatographic studies were performed using a Luna C8(2), 5 μm, 100Å, column (250 × 4.6 mm, Phenomenex) with a mobile phase composed of acetonitrile/2 mM ammonium acetate (51:49 v/v), pH 6.7. The flow rate was 1 mL·min−1 with UV detection at 289 nm. Linearity was observed within the range of 4–36 µg·mL−1 for domperidone and 2–18 µg·mL−1 for lansoprazole. Method optimization was achieved using Box-Behnken design software, in which three key variables were examined, namely, the flow rate (A), the composition of the mobile phase (B), and the pH (C). The retention time (Y1 and Y3) and the peak area (Y2 and Y4) were taken as the response parameters. We observed that slight alterations in the mobile phase and the flow rate influenced the outcome, whereas the pH exerted no effect. Method validation featured various ICH parameters including linearity, limit of detection (LOD), accuracy, precision, ruggedness, robustness, stability, and system suitability. This method is potentially useful for the analysis of commercial formulations and laboratory preparations.