Prediction model for delayed behavior of early ambulation after surgery for varicose veins of lower extremity: a prospective case‒control study.

OBJECTIVE: To analyse influencing factor and establish a prediction model for delayed behavior of early ambulation after surgery for varicose veins of lower extremity (VVLE). DESIGN: A prospective case‒control study. SETTING: VVLE patients were recruited from two local hospitals. PARTICIPANTS: 498 patients with VVLE were selected by convenience sampling and divided into a training set and test set. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Participants were collected information before surgery and followed up until the day after surgery, then divided into a normal and delayed ambulation group. Propensity score matching (PSM) was applied in all participants by type of surgery and anaesthesia. All information of two groups were compared using logistic regression, back propagation neural network (BPNN) and decision tree models. The accuracy, sensitivity, specificity, and area under the curve (AUC) of three models were compared to determine the optimal model. RESULTS: A total of 406 participants were included after PSM. The AUCs of training sets of logistic regression, BPNN and decision tree models were 0.850, 0.932, and 0.757, respectively. The AUCs of test sets were 0.928, 0.984, and 0.776, respectively. BPNN was the optimal model. SSRS score, preoperative 30-second sit-stand test score, CEAP grade, MCMQ score and whether you know the need for early ambulation, in descending order of the result of BPNN model. A probability value greater than 0.56 indicated delayed early ambulation. CONCLUSIONS: Clinicians should pay more attention to those with lower SSRS score, poor lower limb strength, higher CEAP grade and poor medical coping ability, and make patients aware of the necessity and importance of early ambulation, thereby assisting decision-making regarding postoperative rehabilitation. Further research is needed to improve the method, add more variables and transform the model into a scale to screen and intervene in the delay behavior of early ambulation of VVLE in advance.

Characterization of Evodia rutaecarpa (Juss) Benth honey: volatile profile, odor-active compounds and odor properties.

BACKGROUND: Aroma is one of the most important quality criterion of different honeys and even defines their merchant value. The composition of volatile compounds, especially the characteristic odor-active compounds, contributes significantly to the aroma of honey. Evodia rutaecarpa (Juss) Benth honey (ERBH) is a special honey in China with unique flavor characteristics. However, no work in the literature has investigated the volatile compounds and characteristic odor-active compounds of ERBHs. Therefore, it is imperative to conduct systematic investigation into the volatile profile, odor-active compounds and odor properties of ERBHs. RESULTS: The characteristic fingerprint of ERBHs was successfully constructed with 12 characteristic peaks and a similarity range of 0.785-0.975. In total, 297 volatile compounds were identified and relatively quantified by headspace solid-phase microextraction coupled with gas chromatography quadrupole time-of-flight mass spectrometry, of which 61 and 31 were identified as odor-active compounds by relative odor activity values and GC-olfactometry analysis, respectively, especially the common 22 odor-active compounds (E)-ß-damascenone, phenethyl acetate, linalool, cis-linalool oxide (furanoid), octanal, hotrienol, trans-linalool oxide (furanoid), 4-oxoisophorone and eugenol, etc., contributed significantly to the aroma of ERBHs. The primary odor properties of ERBHs were floral, followed by fruity, herbaceous and woody aromas. The partial least-squares regression results showed that the odor-active compounds had good correlations with the odor properties. CONCLUSION: Identifying the aroma differences of different honeys is of great importance. The present study provides a reliable theoretical basis for the quality and authenticity of ERBHs. © 2023 Society of Chemical Industry.

C/H/O stable isotope of rape honey and its components combined with machine learning algorithms to identify geographical origin.

RATIONALE: The distribution of rape honey is among the largest and most diverse of all honeys available to humankind with respect to the geographical origin. Accurate isotopic reference values for rape honey are therefore important for precise verification of honey origin and its traceability. New combined rape honey δ13 C, δ2 H, and δ18 O values in combination with values on its compounds (protein and saccharides) were used to complement existing databases to better identify the geographical origin of Chinese rape honey. METHODS: Traceability methods based on elemental analyzer isotope ratio mass spectrometry and liquid chromatography isotope ratio mass spectrometry were established for geographical origin of rape honey. RESULTS: Rape honey harvested in the high-altitude region (QH; Qinghai) had significantly higher values (1.4 to 5.3‰ for δ13 C, 7.9 to 12.9‰ for δ2 Hprotein ) for the δ13 C of whole honey (-23.8‰), its protein (-24.4‰), fructose (-23.5‰), glucose (-23.6‰), and disaccharide (-24.7‰), and also δ2 H of the protein (103.5‰) than those in low-altitude regions (HB; Hubei, SC; Sicuan, and JS; Jingsu). The δ18 Orape honey was a useful index to differentiate whether rape honey from coastal (JS) or non-coastal (HB, SC, and QH) regions. The δ13 C, δ2 H, and δ18 O values in rape honey are affected by geographical factors, such as temperature and altitude. The δ13 Cprotein and δ13 Crape honey values were better to identify the geographical origin of rape honey than δ13 Csaccharides . The δ18 O and δ2 H values of rape honey protein were more suitable for traceability than those of rape honey. The combination of the δ13 C, δ2 H, and δ18 O values of rape honey and its extracted protein and saccharides improved the precision of three models (linear discriminant analysis, SVM, and random forest) used to discriminate rape honey from different regions in China. The SVM model obtained the best accuracy (93.2%). CONCLUSIONS: Stable isotopes could be significant predictors in determining the geographical origin of rape honey.

GABA negatively regulates adventitious root development in poplar.

γ-Aminobutyric acid (GABA) influences plant growth, but little is known about how this metabolite regulates adventitious root (AR) development. Here, we investigate the effects of GABA on ARs using poplar lines overexpressing glutamate decarboxilase 2 (GAD2) and by treating poplar stem cuttings with exogenous GABA or vigabatrin (VGB; a specific GABA transaminase inhibitor). Endogenous GABA accumulation not only inhibited AR growth, but it also suppressed or delayed AR formation. Anatomical observations revealed that the GABA and VGB treatments resulted in a 1 d delay in the formation of AR primordia and the appearance of ARs. This delay coincided with changes in primary metabolism, including transient increases in hexose and amino acid levels. GABA-dependent changes in the expression of genes related to hormone synthesis and signalling, as well as analysis of hormone levels revealed that ethylene-dependent pathways were decreased at the earliest stage of AR formation. In contrast, auxin and abscisic acid were increased at 1-5 d as well as GA4 over a 5 d period of AR formation. These results demonstrate that GABA plays a crucial role in AR development. Evidence is presented demonstrating that GABA can interact with hormone-related pathways as well as carbon/nitrogen metabolism. These findings also elucidate the functions of GABA in plant development.

Occurrence of Neonicotinoids in Chinese Apiculture and a Corresponding Risk Exposure Assessment.

Neonicotinoids are the most widely used insecticides worldwide, but there is mounting evidence demonstrating that they have adverse effects on nontarget organisms. However, little is known about the extent of environmental neonicotinoids contamination in China. In this study, a total of 693 honey samples from across China, from both Apis melifera and Apis cerana, were analyzed to examine neonicotinoid concentrations and their geographical distribution, and correlation with the primary plant species from which the honey was obtained. Furthermore, chronic and acute exposure risk and risk ranking for humans eating honey were investigated, and risks to bees were also considered. The results revealed that 40.8% of the samples contained at least one of the five neonicotinoids tested. Honeys from commercial crops were found to be more frequently contaminated with neonicotinoids than those from noncommercial crops. Honey samples from Apis mellifera were more frequently contaminated than those from Apis cerana. The concentrations of neonicotinoids found in honey overlapped with those that have been found to have significant adverse effects on honeybee health. The dietary risk assessments indicated that the levels of neonicotinoids detected in honey were likely to be safe for human consumption.

Inhibition of α-ketoglutarate dehydrogenase activity affects adventitious root growth in poplar via changes in GABA shunt.

MAIN CONCLUSION: Blocking α-ketoglutarate dehydrogenase results in up-regulation of γ-aminobutyric acid (GABA) shunt activity, and inhibits the growth of poplar adventitious roots (ARs), indicating that AR growth is closely associated with GABA shunt. γ-Aminobutyric acid (GABA) shunt starts from α-ketoglutarate in the tricarboxylic acid cycle, which is thought to represent the cross road between carbon and nitrogen metabolism. Previous studies (Araújo et al. 2012b, Plant Cell 24: 2328-2351) have shown that blocking α-ketoglutarate dehydrogenase (α-KGDH) affects the GABA shunt activity, and inhibits growth. However, its effects on the growth of adventitious roots (ARs) are unclear. In this study, the growth of ARs in tissue-cultured 84K poplar (Populus alba × Populus glandulosa cv. '84K') was significantly inhibited when succinyl phosphate (SP), a specific inhibitor of α-KGDH, was supplied. The inhibition of ARs was associated with significant changes in the levels of soluble sugars, organic acids, and amino acids, and was coupled with the up-regulation of the GABA shunt activity at the transcriptional and translational levels. Exogenous GABA also inhibited AR growth following the increase of the endogenous GABA level. Transcriptomic analyses further showed that genes related to cell wall carbon metabolism and phytohormone (indoleacetic acid, ABA, and ethylene) signaling were affected by the changes of GABA shunt activity, resulting from the α-KGDH inhibition. Thus, our study indicates that the inhibition of poplar AR growth by blocking α-KGDH is closely associated with GABA shunt, which would benefit a better understanding of GABA's roles in plant development and stress response.

Roles of γ-aminobutyric acid on salinity-responsive genes at transcriptomic level in poplar: involving in abscisic acid and ethylene-signalling pathways.

MAIN CONCLUSION: γ-Aminobutyric acid (GABA) affected ABA and ethylene metabolic genes and signal components in salt-treated poplar, indicating its potential role in signal pathways of ABA and ethylene during salt stress. GABA is a small signalling molecule that accumulates rapidly in plants exposed to various stresses. However, the relationship between GABA and other signalling molecules, such as hormones, remains unclear. Here, in the poplar woody plant under 200-mM NaCl conditions, the application of low (0.25 mM) and high (10 mM) exogenous GABA, compared to 0 mM, affected the accumulation of hydrogen peroxide and hormones, including ABA and ethylene, in different manners. Transcriptomic analysis demonstrated that 1025 differentially expressed genes (DEGs; |log2Ratio| ≥ 1.5) were widely affected by exogenous GABA under salt stress. A clustering analysis revealed that GABA could rescue or promote the effects of salt stress on gene expression. Among them, 146 genes involved in six hormone-signalling pathways were enriched, including 22 ABA- and 50 ethylene-related genes. Quantitative expression of selected genes involved in hormone-related pathways showed that ABA metabolic genes (ABAG, ABAH2, and ABAH4), ethylene biosynthetic genes (ACO1, ACO2, ACO5, ACOH1, ACS1, and ACS7) and receptor genes (PYL1, PYL2, PYL4, and PYL6) were regulated by exogenous GABA, even at a 0.1 mM level. The production of ABA was negatively correlated with ABAH expression levels at different GABA concentrations. The increase of endogenous GABA, resulting from inhibitor (succinyl phosphonate) of α-ketoglutarate dehydrogenase, affected the PYLs levels. Thus, GABA may be involved in ABA- and ethylene-signalling pathways. Our data provide a better understanding of GABA's roles in the plant responses to environmental stresses.

Characterization of Chinese Unifloral Honeys Based on Proline and Phenolic Content as Markers of Botanical Origin, Using Multivariate Analysis.

The phenolic and proline content were determined in honey samples of different floral origins (rapeseed, sunflower, buckwheat and Codonopsis) from five different regions of China. The phenolic and proline profile of these samples were used to construct a statistical model to distinguish honeys from different floral origins. Significant differences were identified among the studied honey samples from multivariate chemometric methods. The proline content varied among the four types of honeys, with the values decreasing in the order: buckwheat > Codonopsis > sunflower > rapeseed. Rapeseed honeys contained a high level of benzoic acid, while rutin, p-coumaric acid, p-hydroxybenzoic acid were present at relatively high levels in buckwheat honeys. Principal component analysis (PCA) revealed that rapeseed honey could be distinguished from the other three unifloral honeys, and benzoic acid, proline and kaempferol could serve as potential floral markers. Using 18 phenolic compounds and proline the honey samples were satisfactorily classified according to floral origin at 94% correct prediction by linear discriminant analysis (LDA). The results indicated that phenolic compounds and proline were useful for the identification of the floral origin of the four type honeys.

[Application of ICP-MS to Identify the Botanic Source of Characteristic Honey in South Yunnan].

By adopting inductively coupled plasma mass spectrometry (ICP-MS) combined with chemometric analysis technology, 23 kinds of minerals in four kinds of characteristic honey derived from Yunnan province were analyzed. The result showed that 21 kinds of mineral elements, namely Na, Mg, K, Ca, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Sr, Mo, Cd, Sb, Ba, Tl and Pb, have significant differences among different varieties of honey. The results of principal component analysis (PCA) showed that the cumulative variance contribution rate of the first four main components reached 77.74%, seven kinds of elements (Mg, Ca, Mn, Co, Sr, Cd, Ba) from the first main component contained most of the honey information. Through the stepwise discriminant analysis, seven kinds of elements (Mg, K, Ca, Cr, Mn, Sr, Pb) were filtered. out and used to establish the discriminant function model, and the correct classification rates of the proposed model reached 90% and 86.7%, respectively, which showed elements contents could be effectively indicators to discriminate the four kinds characteristic honey in southern Yunnan Province. In view of all the honey samples were harvested from apiaries located at south Yunnan Province where have similar climate, soil and other environment conditions, the differences of the mineral elements contents for the honey samples mainly due to their corresponding nectariferous plant. Therefore, it is feasible to identify honey botanical source through the differences of mineral elements.

[A Feasibility Study on the Discrimination of the Propolis Varieties Based on Near Infrared Spectroscopy].

Botanical origins of propolis are significant factors affecting biological and pharmacological activities because of different components in propolis. Until now, the determination of propolis botanical origins is mainly based on different varieties and the content of the compositions with great limitations. Therefore, it is important to discriminate different botanical origins of propolis quickly and accurately. In this study, Near-infrared (NIR) spectra of propolis varieties based on principal component analysis mahalanobis distance (PCA-mahalanobis distance) model and canonical discriminant analysis model were built for the classification of three botanical origins (poplar propolis，brich propolis and rubber propolis). The models were built based on the optimal pretreatment method and bands of first derivative + Savitzky-Golay (7) filter and 4500~12 000-1, which were selected in advance. After the principal component analysis, the correct classification rates of calibration sets and validation sets in analysis mahalanobis distance models were 93.62% and 82.61%, respectively. The discrimination rate and the cross-validation rate of canonical discrimination models were 91.4% and 88.6%, respectively. Therefore, NIR spectroscopy with chemometric methods is not only feasible but also practical for rapid and accurate identification of varieties of propolis.

[Discrimination of Rice Syrup Adulterant of Acacia Honey Based Using Near-Infrared Spectroscopy].

At present, the rice syrup as a low price of the sweeteners was often adulterated into acacia honey and the adulterated honeys were sold in honey markets, while there is no suitable and fast method to identify honey adulterated with rice syrup. In this study, Near infrared spectroscopy (NIR) combined with chemometric methods were used to discriminate authenticity of honey. 20 unprocessed acacia honey samples from the different honey producing areas, mixed? with different proportion of rice syrup, were prepared of seven different concentration gradient? including 121 samples. The near infrared spectrum (NIR) instrument and spectrum processing software have been applied in the? spectrum? scanning and data conversion on adulterant samples, respectively. Then it was analyzed by Principal component analysis (PCA) and canonical discriminant analysis methods in order to discriminating adulterated honey. The results showed that after principal components analysis, the first two principal components accounted for 97.23% of total variation, but the regionalism of the score plot of the first two PCs was not obvious, so the canonical discriminant analysis was used to make the further discrimination, all samples had been discriminated correctly, the first two discriminant functions accounted for 91.6% among the six canonical discriminant functions, Then the different concentration of adulterant samples can be discriminated correctly, it illustrate that canonical discriminant analysis method combined with NIR spectroscopy is not only feasible but also practical for rapid and effective discriminate of the rice syrup adulterant of acacia honey.

[Application of inductively coupled plasma mass spectrometry with chemometric methods in classification of honeys according to their types].

In order to identify honeys according to their floral origin, inductively coupled plasma mass spectrometry (ICP-MS) combined with principal component analysis (PCA) and discriminant analysis (DA) were employed in the present study. Three kinds of honeys such as acacia honey samples, sunflower honey samples and rape honey samples were selected. It was pretreated by wet-acid digestionand measured 20 kinds of mineral elements in honey samples by ICP-MS. The result showed that the accuracy of the inductively coupled plasma mass spectrometrymeted the requirements. The result of principal component analysis demonstrated that the acacia honey samples were performed a trend of certain gather. The trend of the sunflower honey samples and the rape honey samples are not obvious. Ten kinds of mineral elements including Na, Mg, K, Ca, Sr, Ba, V, Fe, Ni, Sb can be regarded as honey varieties of characteristic elements. Seven kinds of mineral elements such as Mg, Sr, Ba, Sb, Ni, Cr and Na could be selected through stepwise discriminant analysis. Using bayes discriminant analysis, A linear discriminant function can be recieved. The discrimination rate of honey samples such as acacia honey samples, sunflower samples and rape honey samples were 100%, 80% and 90. 9% respectively. Two sunflower honey samples was misclassified into rape honey samples an-done rape honey samples are also misclassified into acacia honey sample. The total rate of discriminant model cross validation was 90. 3%. It is concluded that the mineral elements in honey varieties with good classification. The present study can provide theoretical basis and the relationship between thetypes of honey samples with mineral elements. The method what this study used had simple, accurate and stablecharacteristics, which can be used as a reliable method of honey sample identification.

Aromatic profiles and enantiomeric distributions of volatile compounds during the ripening of Dendropanax dentiger honey.

Dendropanax dentiger honey (DDH) is a specialty herbal honey from China. Previous research on DDH has mostly focused on its composition and potential chemical markers, no studies have been conducted on the changes in aroma characteristics and chiral odorants during its maturation. Therefore, the present study aims to address the missing parts. The proportions and total concentrations of 185 volatile compounds identified in different classes varied with DDHs ripening. Fourteen common odor-active compounds were identified by odor activity values (OAVs) and GC-olfactometry (GC-O) analysis. The aroma profiles of DDHs were observed to vary at different ripening stages, although the dominant aroma characteristic was "fruity" aroma, which became more pronounced with increasing maturity. The enantiomeric contents and distributions of 7 volatile enantiomers were related to specific physicochemical indicators and the maturity of DDHs, among which the enantiomers of linalool oxide A may be a potential indicator to identify its maturity. Furthermore, precise quantification and OAVs calculation showed that the enantiomer (2S, 5S)-linalool oxide A presented the highest concentration (8.83-27.39 ng/mL) and only the enantiomer R-linalool (OAVs: 5.56-6.14) was an important contributor to the aroma profiles of DDHs at different stages of maturity. These results provided a new research idea for quality control and identification of DDHs at different maturity stages.

Dynamic variation in the aroma characteristics of Rhus chinensis honey at different stages after capping.

The ripening characteristics after capping of honey are favourable for improving its quality. However, research on the variation and formation of aroma characteristics of honey in this process is lacking. Therefore, the present study was carried out with different stages of Rhus chinensis honeys (RCHs) after capping and identified 192 volatiles with varying levels of concentration. "Fruity" was the main aroma characteristic of RCHs at different stages after capping, mainly contributed by (E)-ß-damascenone. Methyl salicylate might be a potential indicator for differentiating RCHs at different stages after capping. The metabolic pathway analyses revealed that the aroma compounds in RCHs undergo transformation at different stages after capping, which subsequently affects its aroma characteristics formation. This work is the first to study the dynamic changes in honey aroma characteristics after capping from multiple perspectives, and the results are of great significance for understanding the aroma characteristics after capping and quality control of honey.

Effects of different floral periods and environmental factors on royal jelly identification by stable isotopes and machine learning analyses during non-migratory beekeeping.

It is crucial to monitor the authenticity of royal jelly (RJ) because the qualities of RJs produced by different floral periods vary substantially. In the context of non-migratory beekeeping, this study aims to identify rape RJ (RRJ), chaste RJ (CRJ), and sesame RJ (SRJ) based on δ13C, δ15N, δ2H, and δ18O combined with machine learning and to evaluate environmental effect factors. The results showed that δ13C (-27.62‰ ± 0.24‰), δ15N (2.88‰ ± 0.85‰), and δ18O (28.02‰ ± 1.30‰) of RRJ were significantly different from other RJs. The δ13C, δ2H, and δ18O in CRJ and SRJ were strongly correlated with temperature and precipitation, suggesting that these isotopes are influenced by environmental elements such as sunlight and rainfall. In addition, the artificial neural network (ANN) model was superior to the random forest (RF) model in terms of accuracy, sensitivity, and specificity. This study revealed that combining stable isotopes with ANN models and the unique correlation between stable isotopes and environmental factors could provide promising ideas for monitoring the authenticity of RJ.

Analytical Strategies for LC-MS-Based Untargeted and Targeted Metabolomics Approaches Reveal the Entomological Origins of Honey.

A comprehensive liquid chromatography-mass spectrometry (LC-MS)-based metabolomics approach was developed to discriminate honey harvested from Apis mellifera ligustica Spinola (A. mellifera) and Apis cerana cerana Fabricius (A. cerana). Based on an untargeted strategy, ultrahigh-performance liquid chromatography electrospray ionization quadrupole orbitrap high-resolution mass spectrometry (UPLC Q-Orbitrap) was combined with chemometrics techniques to screen and identify tentative markers from A. mellifera and A. cerana honey. In targeted metabolomics analysis, a sensitive method of solid-phase extraction followed by ultrahigh-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UPLC-MS/MS) was established for quantifying three markers, and the results showed that 3-amino-2-naphthoic acid and methyl indole-3-acetate could be considered markers of A. cerana honey, as they were present in higher amounts in A. cerana honey than in A. mellifera honey, whereas kynurenic acid was determined to be a marker of A. mellifera honey. This work highlights critical information for the authentication of A. cerana and A. mellifera honey.

Study on stable carbon isotope fractionation of rape honey from rape flowers (Brassica napus L.) to its unifloral ripe honey.

A new idea and strategy for honey traceability and identification was provided by studying the carbon isotope fractionation of rape honey and its components in the different ripening process, as well as the fractionation from rape flowers, stamens, nectar to rape honey. The results showed the moisture content of rape honey continued to decrease, and the glucose and fructose content continued to increase during the ripening process. The δ13C of rape honey and its protein were less affected by honey ripeness, while the δ13C of sugars in rape honey were greatly affected by this. At the same time, the fractionation of carbon isotope from rape flowers to honey was significant. The δ13C of rape honey and its protein, disaccharide, fructose, and glucose had a strong correlation, and the δ13C of rape honey and its components were mainly related to rape flowers and its stamens.

The landscape of COVID-19 vaccination among healthcare workers at the first round of COVID-19 vaccination in China: willingness, acceptance and self-reported adverse effects.

The COVID-19 vaccines have been developed in a wide range of countries. This study aimed to examine factors that related to vaccination rates and willingness to be vaccinated against COVID-19 among Chinese healthcare workers (HCWs). From 3rd February to 18th February, 2021, an online cross-sectional survey was conducted among HCWs to investigate factors associated with the acceptance and willingness of COVID-19 vaccination. Sociodemographic characteristics and the acceptance of COVID-19 vaccination among Chinese HCWs were evaluated. A total of 2156 HCWs from 21 provinces in China responded to this survey (effective rate: 98.99%)), among whom 1433 (66.5%) were vaccinated with at least one dose. Higher vaccination rates were associated with older age, working as a clinician, having no personal religion, working in a fever clinic or higher hospital grade, and having received vaccine education, family history for influenza vaccination and strong familiarity with the vaccine. Willingness for vaccination was related to working in midwestern China, considerable knowledge of the vaccine, received vaccine education, and strong confidence in the vaccine. Results of this study can provide evidence for the government to improve vaccine coverage by addressing vaccine hesitancy in the COVID-19 pandemic and future public health emergencies.

Quantification of melittin and apamin in bee venom lyophilized powder from Apis mellifera by liquid chromatography-diode array detector-tandem mass spectrometry.

A high-performance liquid chromatography-diode array detector-tandem mass spectrometry (HPLC-DAD-MS/MS) method was developed for simultaneous determination of melittin and apamin in crude bee venom lyophilized powder (CBVLP) as the traditional Chinese medicine possessing specific biological activity. Melittin and apamin were extracted with pure water from CBVLP samples followed by HPLC-DAD-MS/MS analysis. The method was validated to demonstrate its selectivity, linearity, limit of quantification (LOQ), intraday precision, interday precision, accuracy, recovery, matrix effect, and stability. The assay was linear over the concentration ranges of 1-100 and 0.2-25 microg/ml with limit of quantifications (LOQs) of 1.0 and 0.3 microg/ml for melittin and apamin, respectively. The precision results were expressed as coefficients of variation (CVs), ranging from 2.2% to 11.4% for intraday repeatability and from 3.2% to 13.1% for interday intermediary precision. The concentrations of endogenous melittin and apamin in CBVLP samples ranged from 46% to 53% and from 2.2% to 3.7% of dry weight, respectively. This rapid, simple, precise, and sensitive method allowed the simultaneous determination of melittin and apamin to evaluate authenticity and quality of CBVLP samples.

Determination of melamine in royal jelly and lyophilized powder using ultrasonically assisted extraction by ion-pair HPLC with UV detection.

A rapid method for the analysis of melamine in royal jelly (RJ) and RJ lyophilized powder (RJLP) was developed using ion-pair RP-HPLC coupled with UV detector. The method utilized an optimized buffer system to avoid the elution of melamine near the column void volume and improve retention of melamine in a generic C8 chromatographic column. In addition, sample preparation included deproteination, ultrasonic-assisted extraction, and cleanup on a mixed-mode cation exchange extraction cartridge. The extraction procedure was optimized with regard to the amount of extraction solvent and the duration of sonication for RJ and RJLP samples. The following criteria were used to validate the HPLC-UV detection method: selectivity, linearity, precision, LOD, and LOQ. Correlation coefficient was higher than 0.999 by applying the linear regression model based on the least square method with a weighting factor (1/x). Precision was evaluated as repeatability and intermediary precision with RSD of less than 15%. The mean percentage recoveries of melamine were varied from 72.5 to 90.5% for RJ and RJLP. This approach will be of particular utility for the evaluation of melamine residue level and routine monitoring of melamine in RJ and RJLP samples.