In this work the feasibility of near infrared spectroscopy was evaluated combined with chemometric approaches, as a tool for the botanical origin prediction of 119 honey samples. Four varieties related to polyfloral, acacia, chestnut, and linden were first characterized by their physical-chemical parameters and then analyzed in triplicate using a near infrared spectrophotometer equipped with an optical path gold reflector. Three different classifiers were built on distinct multivariate and machine learning approaches for honey botanical classification. A partial least squares discriminant analysis was used as a first approach to build a predictive model for honey classification. Spectra pretreatments named autoscale, standard normal variate, detrending, first derivative, and smoothing were applied for the reduction of scattering related to the presence of particle size, like glucose crystals. The values of the descriptive statistics of the partial least squares discriminant analysis model allowed a sufficient floral group prediction for the acacia and polyfloral honeys but not in the cases of chestnut and linden. The second classifier, based on a support vector machine, allowed a better classification of acacia and polyfloral and also achieved the classification of chestnut. The linden samples instead remained unclassified. A further investigation, aimed to improve the botanical discrimination, exploited a feature selection algorithm named Boruta, which assigned a pool of 39 informative averaged near infrared spectral variables on which a canonical discriminant analysis was assessed. The canonical discriminant analysis accounted a better separation of samples according to the botanical origin than the partial least squares discriminant analysis. The approach used has permitted to achieve a complete authentication of the acacia honeys but not a precise segregation of polyfloral ones. The comparison between the variables important in projection and the Boruta pool showed that the informative wavelengths are partially shared especially in the middle and far band of the near infrared spectral range.
This study aimed to define the pharmacokinetic profiles of dexmedetomidine and methadone administered simultaneously in dogs by either an oral transmucosal route or intramuscular route and to determine the bioavailability of the oral transmucosal administration relative to the intramuscular one of both drugs, so as the applicability of this administration route in dogs. Twelve client‐owned dogs, scheduled for diagnostic procedures, were treated with a combination of dexmedetomidine hydrochloride (10 μg/kg) and methadone hydrochloride (0.4 mg/kg) through an oral transmucosal route or intramuscularly. Oral transmucosal administration caused ptyalism in most subjects, and intramuscular administration caused transient peripheral vasoconstriction. The results showed reduced and delayed absorption of both dexmedetomidine and methadone when administered through an oral transmucosal route, with median (range) Cmax values of 0.82 (0.42–1.49) ng/ml and 13.22 (2.80–52.30) ng/ml, respectively. The relative bioavailability was low: 16.34% (dexmedetomidine) and 15.5% (methadone). Intramuscular administration resulted in a more efficient absorption profile, with AUC and Cmax values for both drugs approximately 10 times higher. Dexmedetomidine and methadone administered simultaneously by an oral transmucosal route using injectable formulations were not well absorbed through the oral mucosa. Nevertheless, additional studies on these drugs combination using alternative administration routes are recommended.
Milk proteins genetic variants have always attracted a lot of interest as they are associated with important issues relating to milk composition and technological properties. An important debate has recently opened at an international level on the role of β-casein (β-CN) A1 and A2 polymorphisms, toward human health. For this reason, a lot of efforts has been put into the promotion of A2 milk by companies producing and selling A1-free milk, leading the farmers and breeders to switch toward A2 milk production without paying attention on the potential effect of the processability of milk into cheese. The aim of the present work was to evaluate the effects of β-CN, specifically the A1 and A2 allelic variants, on the detailed milk protein profile and cheese-making traits in individual milk samples of 1,133 Holstein Friesian cows. The protein fractions were measured with reversed-phase (RP)-HPLC (expressed in g/L and % N), and the cheese-making traits, namely milk coagulation properties, cheese yield, and curd nutrient recoveries assessed at the individual level, with a nano-scale cheese-making procedure. The β-CN (CSN2), κ-CN (CSN3), and β-lactoglobulin (LGB) genetic variants were first identified through RP-HPLC and then confirmed through genotyping. Estimates of the effects of protein genotypes were obtained using a mixed inheritance model that considered, besides the standard nuisance variables (i.e., days in milk, parity, and herd-date), the milk protein genes located on chromosome 6 (CSN2, CSN3) and on chromosome 11 (LGB), and the polygenic background of the animals. Milk protein genes (CSN2, CSN3, and LGB) explained an important part of the additive genetic variance in the traits evaluated. The β-CN A1A1 was associated with a significantly lower production of whey proteins, particularly of β-lactoglobulin (−8.2 and −6.8% for g/L and % N, respectively) and α-lactalbumin (−4.7 and −4.4% for g/L and % N, respectively), and a higher production of β-CN (6.8 and 6.1% for g/L and % N, respectively) with respect to the A2A2 genotype. Regarding milk cheese-making ability, the A2A2 genotype showed the worst performance compared with the other genotypes, particularly with respect to the BA1, with a higher rennet coagulation time (7.1 and 28.6% compared with A1A1 and BA1, respectively) and a lower curd firmness at 30 min. Changes in milk protein composition through an increase in the frequency of the A2 allele in the production process could lead to a worsening of the coagulation and curd firming traits.
Toxic pyrrolizidine alkaloids (PAs) and their N-oxides (PANOs) can be present in bee pollen depending on the plants visited by bees. A liquid chromatography-mass spectrometry (LC-MS/MS) method was developed and validated to monitor 17 PAs/PANOs in 44 bee pollens. The CIE-L*a*b* colour coordinates with the specular component either included or excluded were recorded in pellets and ground aliquots. Lightness (L*) and yellowness (b*) of ground bee pollen were significantly correlated to PAs/PANOs content. The L* and b* cut-offs sorted by a receiver operating characteristic analysis to predict PAs/PANOs presence showed a significant increase in the relative risk to detect amounts higher than 84 μg kg À1 . Two supervised canonical discriminant analyses confirmed that pollen without PAs could be distinguished from those containing PAs/PANOs. The data suggest that instrumental colour coupled with supervised models could be used as a screening test for PAs/PANOs in bee pollen, before the confirmatory LC-MS/MS analysis.
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