Procedure to explore a ternary mixture diagram to find the appropriate gradient profile in liquid chromatography with fluorescence detector. Application to determine four primary aromatic amines in napkins.

The purpose of this work is to develop a tool to search for a gradient profile with ternary or binary mixtures in liquid chromatography, that can provide well-resolved chromatograms in the shortest time for multianalyte analysis. This approach is based exclusively on experimental data and does not require a retention time model of the compounds to be separated. The methodology has been applied for the quantification of four primary aromatic amines (PAAs) using HPLC with fluorescence detector (FLD). Aniline (ANL), 2,4-diaminotoluene (TDA), 4,4'-methylenedianiline (MDA) and 2-aminobiphenyl (ABP) have been selected since their importance in food contact materials (FCM). In order to achieve that, partial least squares (PLS) models have been fitted to relate CMP (control method parameters) and CQA (critical quality attributes). Specifically, PLS models have been fitted using 30 experiments for each one of the four CQA (resolution between peaks and total elution time), considering 33 predictor variables (the composition of the methanol and acetonitrile in the mobile phase and the time of each one of the 11 isocratic segments of the gradient). These models have been used to predict new candidate gradients, and then, some of those predictions (the ones with resolutions above 1.5, in absolute value, and final time lower than 20 min) have been experimentally validated. Detection capability of the method has been evaluated obtaining 1.8, 189.4, 28.8 and 3.0 µg L-1 for ANL, TDA, MDA and ABP, respectively. Finally, the application of chemometric tools like PARAFAC2 allowed the accurate quantification of ANL, TDA, MDA and ABP in paper napkins in the presence of other interfering substances coextracted in the sample preparation process. ANL has been detected in the three napkins analysed in quantities between 33.5 and 619.3 µg L-1, while TDA is present in only two napkins in quantities between 725.9 and 1908 µg L-1. In every case, the amount of PAAs found, exceeded the migration limits established in European regulations.

Method operable design region obtained with a partial least squares model inversion in the determination of ten polycyclic aromatic hydrocarbons by liquid chromatography with fluorescence detection.

A chromatographic method with the Analytical Quality by Design (AQbD) methodology is developed for the simultaneous determination by HPLC-FLD of ten PAHs (naphthalene, phenanthrene, anthracene, fluoranthene, pyrene, chrysene, benzo[a]anthracene, perylene, benzo[b]fluoranthene, and benzo[a]pyrene), widely spread in the environment. The construction of the Method Operable Design Region (MODR) is conducted, for the first time, via the inversion of a multiresponse Partial Least Squares (PLS2) model, which is needed to maintain the correlations among the Critical Method Parameters (CMP), among the Critical Quality Attributes (CQA), and the covariance between one another. The five CMP considered were the composition of the mobile phase (water, methanol, acetonitrile), flow rate, and column temperature. The eight CQA were linked to resolution between peaks recorded in the same emission wavelength (greater than 1.4) and the total time (less than 15 minutes). By systematic use of experimental design and parallel coordinates plots to explore the Pareto optimal front obtained with the PLS2 model inversion, the computed MODR is formed by convex combinations of eight specific settings of Critical Method Parameters that have a mobile phase with percentages of water between 37 and 38 %, of methanol from 13 and 22 %, and of acetonitrile between 41 and 49 %, together with a flow rate between 1.47 and 1.50 mL min-1, and column temperature between 41.9 and 44.0 °C in their adequate combinations. All the chromatographic peaks are well resolved, with total time varying between 12.96 and 15.66 min inside the estimated MODR and the analytical method is accurate with CCß between 0.9 and 7.0 µg L-1 with probability of both false positive and false negative equal to 0.05.

A new approach based on inversion of a partial least squares model searching for a preset analytical target profile. Application to the determination of five bisphenols by liquid chromatography with diode array detector.

The paper shows a procedure for selecting the control method parameters (factors) to obtain a preset 'analytical target profile' when a liquid chromatographic technique is going to be carried out for the simultaneous determination of five bisphenols (bisphenol-A, bisphenol-S, bisphenol-F, bisphenol-Z and bisphenol-AF), some of them regulated by the European Union. The procedure has three steps. The first consists of building a D-optimal combined design (mixture-process design) for the control method parameters, which are the composition of the ternary mobile phase and its flow rate. The second step is to fit a PLS2 model to predict six analytical responses (namely, the resolution between each pair of consecutive peaks, and the initial and final chromatographic time) as a function of the control method parameters. The third final step is the inversion of the PLS2 model to obtain the conditions needed for attaining a preset analytical target profile. The computational inversion of the PLS2 prediction model looking for the Pareto front of these six responses provides a set of experimental conditions to conduct the chromatographic determination, specifically 22% of water, mixed with 58% methanol and 20% of acetonitrile, keeping the flow rate at 0.66 mL min-1. These conditions give a chromatogram with retention times of 2.180, 2.452, 2.764, 3.249 and 3.775 min for BPS, BPF, BPA, BPAF and BPZ, respectively, and excellent resolution among all the chromatographic peaks. Finally, the analytical method is validated under the selected experimental conditions, in terms of trueness and precision. In addition, the detection capability for the five bisphenols were: 596, 334, 424, 458 and 1156 µg L-1, with probabilities of false positive and of false negative equal to 0.05.

Kinetic models of migration of melamine and formaldehyde from melamine kitchenware with data of liquid chromatography.

European legislation has established a specific migration limit (SML) of 15 mg kg-1 for formaldehyde and 2.5 mg kg-1 for melamine. Formaldehyde resins are used in the manufacture of melamine kitchenware. Formaldehyde is listed in group 1 of the IARC list of carcinogenic compounds. To determine the quantity of formaldehyde and melamine as potential migrants from different types of melamine kitchenware (glass, mug, cutlery, big cup and bowl), a HPLC-DAD method has been implemented. This method is an alternative to the ones proposed in technical guidelines to determine formaldehyde by UV-vis spectrophotometry and melamine by HPLC. The final objective was to fit the migration kinetic curves of these two analytes in melamine kitchenware. After the method was validated, decision limit (CCα) and detection capability (CCß) were calculated for both analytes, when the probabilities of false positive (α) and false negative (ß) were fixed at 0.05; being CCß 0.269 mg L-1 and 0.311 mg L-1 for melamine and formaldehyde respectively. CCα and CCß were also calculated at the SML of both analytes. The migration testing were conducted with simulant B (3% acetic acid (w/v) in aqueous solution), the conditions of each exposure being 70 °C for 2 h. The quantities of melamine and formaldehyde found in the third exposure of the total kitchenware analysed were between 0.21 and 1.09 mg L-1 and between 0.55 and 3.86 mg L-1, respectively. Migration kinetic curves were built for each type of kitchenware with the data of sixteen consecutive migration cycles (70 °C each 30 min). The SML for melamine was surpassed in the mug, in the big cup and in the bowl after eleven, thirteen and one cycles, respectively. When more cycles were carried out in the mug, the values of the accumulated quantity of formaldehyde and melamine were 15.30 and 6.79 mg L-1, respectively, after thirty-two cycles. Both concentrations exceeded the corresponding SML.

Easy-to-use procedure to optimise a chromatographic method. Application in the determination of bisphenol-A and phenol in toys by means of liquid chromatography with fluorescence detection.

Legal limits for phenol and bisphenol-A (BPA) in toys are 15 and 0.1 mg L-1 respectively. The latest studies show that in Europe the content of BPA, which reaches our bodies through different contact routes, in no cases exceed legal limits. But it is true that the effects caused by continued intake of this analyte for a long time and other possible processes that could increase their migration, are still under consideration by the health agencies responsible. A multiresponse optimization using a D-optimal design for simultaneously optimising two experimental factors (temperature and flow) at three levels and one (mobile phase composition) at four levels, in the determination by means of HPLC-FLD is proposed in this work. The D-optimal design allows ones to reduce the experimental effort from 36 to 11 experiments guaranteeing the quality of the estimates. The model fitted is validated and, after the responses are estimated in the whole experimental domain, the experimental conditions that maximize peak areas and minimize retention times for both analytes are chosen by means of a Pareto front. In this way, the sensitivity and the time of the analysis have been improved with this optimization. Decision limit and capability of detection at the limits obtained were 33.9 and 66.1 µg L-1 for phenol and 25.6 and 50.0 for BPA µg L-1 respectively when the probabilities of false negative and false positive were fixed at 0.05. The procedure has been successfully applied to determine phenol and BPA in different samples (toys, clinical serum bags and artificial tears). The simulants HCl 0.07 M and water were used for the analysis of toys. The quantity of phenol found in serum bags and in artificial tears ranged from 15 to 600 µg L-1. No BPA has been found in the objects analysed. In addition, this work incorporates computer programmes which implement the procedure used (COOrdinates parallel plot and Pareto FROnt, COO-FRO) such that it can be used in any other chromatographic optimization.

Influence of gender on carcass yield of twin lambs Blackbelly x Pelibuey at 56 days of age / Influência do gênero no rendimento de carcaça em cordeiros gêmeos Blackbelly x Pelibuey aos 56 días de idade

The aim of this study was to determine the influence of gender on pre-weaning growth, yield and weight of primal cuts of the carcass of Blackbelly x Pelibuey lambs. 16 twin lambs were used. The lamb gender was the fixed factor. The variables evaluated were: body weight at birth and at weaning, average daily weight gain, weight and yield of carcass and primal cuts: neck, arm, thorax, abdomen and leg. Gender did not affect (P>0.05) body weight at birth. However, the pre-weaning average daily weight gain and body weight at weaning were affected (P<0.05) by the gender. Hot carcass yield and weight and primal cuts were not affected (P>0.05) by the gender of the lamb. Nevertheless, the area of the Longissimus dorsi muscle and cavity fat were different between males and females (P<0.05) 11.5±0.73cm2 and 198±0.05 g vs 9.3±0.73cm2 and 282±0.05g, respectively. In conclusion, in twin lambs Blackbelly x Pelibuey males had greater average daily weight gain and body weight at weaning than females. Carcass yield and weight and primal cuts were not affected by the gender of the lamb. However, males had greater area of the L. dorsi muscle than females and these in turn had a greater amount of cavity fat than males.(AU)

Objetivou-se determinar a influência do gênero no crescimento pré-desmame, no rendimento e no peso dos cortes primários de carcaça de cordeiros Blackbelly x Pelibuey. Utilizaram-se 16 cordeiros gêmeos. Os dados foram analisados em um desenho completamente ao acaso, em que o gênero dos corderos foi o fator fixo. As variáveis avaliadas foram: peso vivo ao nascimento e ao desmame, ganho diário de peso, peso e rendimento de carcaças e dos cortes primários: pescoço, braço, tórax, abdômen e perna. O gênero não afetou (P>0,05) o peso vivo ao nascimento. No entanto, o ganho diário de peso e o peso vivo ao desmame foram afetados, sendo esses valores superiores nos machos (P<0,05). O peso e o rendimento das carcaças quentes e dos cortes primários não foram afetados (P>0,05) pelo gênero do cordeiro. Entretanto, a área do músculo Longissimus dorsi e a gordura cavitária foram diferentes entre machos e fêmeas (P<0,05): 11.5±0.73cm2 e 198±0.05g vs. 9.3±0.73cm2 e 282±0.05g, respectivamente. Em conclusão, nos cordeiros gêmeos Blackbelly x Pelibuey, os machos tiveram maior ganho diário de peso e maior peso vivo ao desmame do que as fêmeas. O peso e o rendimento da carcaça bem como os cortes primários não foram afetados pelo gênero do cordeiro. Todavia, os machos tiveram maior área do músculo L. dorsi que as fêmeas e estas tiveram maior quantidade de gordura cavitária que os machos.(AU)

Genetic and non-genetic effects on productive and reproductive traits of cows in dual-purpose herds in southeastern Mexico.

Data on productive and reproductive performance of cows in dual-purpose herds were analyzed to determine the effect of some environmental and genetic factors on saleable milk yield (SMY), lactation length (LL), daily saleable milk per lactation (DMYL), calving interval (CI), and daily saleable milk per calving interval (MYCI) in dual-purpose herds in Yucatan, Mexico. Repeatabilities (re) for these traits were also estimated. Data were obtained from monthly visits to 162 herds from 1996 to 2000. The fixed factors studied were: region (central, eastern and southern), parity number (1 to 6), calving year (1996 to 2000) and calving season (dry, rainy and windy and rainy), genetic group (low- (< 0.50%), medium- (50%) and high- (> 50%) grade cows with European genes). About 2700 to 7700 cows were evaluated for each trait. All factors had significant effects (P < 0.05) on the traits except for region on CI and calving season on DMYL. The overall means for SMY, LL, DMYL, CI, and MYCI were: 1322.3 +/- 80.5 kg, 224.8 +/- 1.3 days, 5.8 +/- 0.1 kg, 555.1 +/- 16.5 days, and 3.0 +/- 0.1 kg, respectively. The re values for SMY, LL, DMYL, CI, and MYCI traits were: 0.19 +/- 0.03, 0.08 +/- 0.04, 0.16 +/- 0.04, 0.00 +/- 0.08, and 0.08 +/- 0.07, respectively. First parity cows had lower SMY, shorter LL, longer CI, and lower MYCI means than cows with more than one parity. Medium grade cows produced more SMY, DMYL and MYCI and had shorter CI than low- and high-grade cows. Therefore, under Yucatan conditions medium-grade cows should be exploited, and more attention should be given to first parity cows in order to improve the productivity in the herd. The relatively high re estimates for SMY and DMYL can be used to calculate most probable producing abilities, in order to identify which cows should be culled.

Genetic and non-genetic effects on productive and reproductive traits of cows in dual-purpose herds in southeastern Mexico

Data on productive and reproductive performance of cows in dual-purpose herds were analyzed to determine the effect of some environmental and genetic factors on saleable milk yield (SMY), lactation length (LL), daily saleable milk per lactation (DMYL), calving interval (CI), and daily saleable milk per calving interval (MYCI) in dual-purpose herds in Yucatan, Mexico. Repeatabilities (re) for these traits were also estimated. Data were obtained from monthly visits to 162 herds from 1996 to 2000. The fixed factors studied were: region (central, eastern and southern), parity number (1 to 6), calving year (1996 to 2000) and calving season (dry, rainy and windy and rainy), genetic group (low- (< 0.50%), medium- (50%) and high- (> 50%) grade cows with European genes). About 2700 to 7700 cows were evaluated for each trait. All factors had significant effects (P < 0.05) on the traits except for region on CI and calving season on DMYL. The overall means for SMY, LL, DMYL, CI, and MYCI were: 1322.3 +/- 80.5 kg, 224.8 +/- 1.3 days, 5.8 +/- 0.1 kg, 555.1 +/- 16.5 days, and 3.0 +/- 0.1 kg, respectively. The re values for SMY, LL, DMYL, CI, and MYCI traits were: 0.19 +/- 0.03, 0.08 +/- 0.04, 0.16 +/- 0.04, 0.00 +/- 0.08, and 0.08 +/- 0.07, respectively. First parity cows had lower SMY, shorter LL, longer CI, and lower MYCI means than cows with more than one parity. Medium grade cows produced more SMY, DMYL and MYCI and had shorter CI than low- and high-grade cows. Therefore, under Yucatan conditions medium-grade cows should be exploited, and more attention should be given to first parity cows in order to improve the productivity in the herd. The relatively high re estimates for SMY and DMYL can be used to calculate most probable producing abilities, in order to identify which cows should be culled.