Performance evaluation of E-nose and E-tongue combined with machine learning for qualitative and quantitative assessment of bear bile powder.

Bear bile powder (BBP) is a valuable animal-derived product with a huge adulteration problem on market. It is a crucially important task to identify BBP and its counterfeit. Electronic sensory technologies are the inheritance and development of traditional empirical identification. Considering that each drug has its own specific odor and taste characteristics, electronic tongue (E-tongue), electronic nose (E-nose) and GC-MS were used to evaluate the aroma and taste of BBP and its common counterfeit. Two active components of BBP, namely tauroursodeoxycholic acid (TUDCA) and taurochenodeoxycholic acid (TCDCA) were measured and linked with the electronic sensory data. The results showed that bitterness was the main flavor of TUDCA in BBP, saltiness and umami were the main flavor of TCDCA. The volatiles detected by E-nose and GC-MS were mainly aldehydes, ketones, alcohols, hydrocarbons, carboxylic acids, heterocyclic, lipids, and amines, mainly earthy, musty, coffee, bitter almond, burnt, pungent odor descriptions. Four different machine learning algorithms (backpropagation neural network, support vector machine, K-nearest neighbor, and random forest) were used to identify BBP and its counterfeit, and the regression performance of these four algorithms was also evaluated. For qualitative identification, the algorithm of random forest has shown the best performance, with 100% accuracy, precision, recall and F1-score. Also, the random forest algorithm has the best R2 and the lowest RMSE in terms of quantitative prediction.

Identification of the Authenticity and Geographical Origin of Bear Bile Powder by Using High Performance Liquid Chromatography - Charged Aerosol Detector Fingerprints Combined with Chemometrics.

Bear bile powder (BBP) is a rare animal-derived traditional Chinese medicine, and it has been widely used to treat visual disorders and hepatobiliary diseases in East Asia. However, there is still a lack of reliable quality control methods for BBP. This study was designed to establish a comprehensive quality map of BBP based on bile acids. High-performance liquid chromatography coupled with charged aerosol detector (HPLC-CAD) was used for fingerprint establishment and quantitative analysis of BBP. The similarities of HPLC-CAD chromatograms for 50 batches of BBP were more than 0.95, while the similarities of reference chromatograms between 6 other animal bile and BBP were low than 0.7. Additionally, five bile acids in BBP, including tauroursodeoxycholic acid, taurocholic acid, taurochenodeoxycholic acid, ursodesoxycholic acid, and chenodeoxycholic acid, were simultaneously quantified. This method has been validated with good regression as well as satisfactory precision, sensitivity, stability, repeatability, and accuracy. Using this method, the contents of five bile acids in BBP samples from five producing areas were determined and compared. Furthermore, Fisher linear discriminant analysis was performed to discriminate the geographic origins of BBP. The result demonstrated that HPLC-CAD fingerprint combined with multi-components quantification is an effective and reliable method for quality control of BBP, it could be a meaningful reference for the quality evaluation of medicinal bile.

Rapid discrimination of the authenticity and geographical origin of bear bile powder using stable isotope ratio and elemental analysis.

Bear bile powder (BBP) is one of the most famous traditional Chinese medicines derived from animals. It has a long history of medicinal use and is widely used in the treatment of hepatobiliary and ophthalmic diseases. Due to its similar morphological characterizations and chemical composition compared with other bile powders, it is difficult to accurately identify its authenticity. In addition, there are very few methods that could analyze the geographical origins of BBP. In this study, elemental analysis isotope ratio mass spectrometry (EA-IRMS) and inductively coupled plasma mass spectrometry (ICP-MS) were used to determine stable isotope ratios and elemental contents, respectively. Combined these variables with chemometrics, the discrimination models were established successfully for identifying the authenticity and geographical origins of BBP. Meanwhile, the discrimination markers were identified by calculating the variable importance for the projection (VIP) value of each variable. A total of 13 discrimination markers (δ13C, δ15N, C, Li, Mg, K, Ca, Cr, Ni, Zn, As, Se, and Sr) were used to further establish the fingerprint of BBP. According to similarity analysis, the authenticity and geographical origins of BBP could be identified without chemometrics. In conclusion, the present study established a reliable method for authenticity identification and origin traceability of BBP, which will provide references for the quality control of bile medicines.

Identification and Quality Evaluation of Velvet Antler by DNA Barcoding and Stable Isotope Techniques Combined with Chemometrics.

This study aimed to identify Velvet antler and its counterfeits and to further evaluate their quality. Mitochondrial cytochrome b (Cytb) was used as a target gene to identify Velvet antler samples, and a DNA barcoding method was established for species origin identification in Velvet antlers. After identification, the stable isotope contents and ratios were adopted to evaluate the quality of different specifications of authentic Velvet antler in combination with chemometrics. Two stable isotope contents (C % and N %) and ratios (δ13C and δ15N) in three kinds of Velvet antler slices of different specifications, namely, wax slices, powder slices, and bone slices, were determined. Nine Velvet antler samples sold in the market were identified for label conformity. Only two samples were consistent with the labeled species, and the others were counterfeits. The three slices of Velvet antler of different specifications were clearly distinguished by principal component analysis and hierarchical cluster analysis. Then, the discriminant model of partial least squares discriminant analysis was established, and 100% discrimination accuracy was observed in this model. All the Velvet antler slice samples of different specification samples were grouped clearly according to their sources. In summary, it is feasible for the identification and quality grade evaluation of Velvet antler by DNA barcoding based on mitochondrial Cytb and stable isotope techniques combined with chemometric analysis. The establishment of this method also provided a reference for the evaluation of other animal-derived medicinal materials.

Purified ß-glucans of Different Molecular Weights Enhance Growth Performance of LPS-challenged Piglets via Improved Gut Barrier Function and Microbiota.

This study investigated ß-glucan derived from Agrobacterium sp. ZX09 with high (2000 kDa) and low (300 kDa) molecular weight (MW) to compare their effects on growth performance and gut function in LPS-induced weaned piglets. Changes in jejunal morphology, mucosal barrier function, microbial populations, and fermentation in the piglets were determined. Data showed that ß-glucan prevented body weight loss in LPS challenged piglets. Supplementation with both ß-glucan fractions improved jejunal morphology. Compared to low MW, ß-glucan of high MW generally up-regulated transcripts of ZO-1, MUC1, and MUC2 in jejunal mucosa to a lesser extent. Mucosal D-lactate, diamine oxidase, and anti-oxidation index were effectively resumed in ß-glucan treatment. Both ß-glucan diets provoked the emergence of a balanced microbiota and a richer concentration of volatile fatty acids in the colon. The richest community of bifidobacterium and concentration of butyrate emerged after feeding ß-glucan with high MW. Results suggested that the effect of Agrobacterium sp. ZX09 ß-glucans on the gut-modulatory function is largely linked to their MW. Low MW ß-glucan mainly improved the mucosal barrier function in the jejunum, while high MW ß-glucan had profound effects on the microbial community and fermentation in the hindgut of piglets.

Dietary ß-glucan supplementation improves growth performance, carcass traits and meat quality of finishing pigs.

This experiment was conducted to investigate growth performance, carcass traits and meat quality of finishing pigs with dietary ß-glucan supplementation. A total of 96 healthy pigs (Duroc × Landrace × Yorkshire; initial average BW = 25 kg) were randomly allocated into 4 dietary treatments with 6 replicates per treatment and 4 pigs per replicate. The control group was fed a basal diet, and the experimental diets were supplemented with 50, 100 and 200 mg/kg Agrobacterium sp. ZX09 ß-glucan, respectively. The experiment lasted 103 d. The basal diet supplemented with 100 mg/kg ß-glucan significantly increased average daily gain and feed conversion ratio, probably due to the improved digestibility of dry matter, gross energy and crude protein (P < 0.05). Beta-glucan supplementation from 100 to 200 mg/kg of diet significantly increased carcass length (P < 0.05). The basal diet supplemented with 100 mg/kg ß-glucan supplementation also significantly (P < 0.05) increased muscle pH, reduced drip losses and increased a∗ values. The basal diet supplemented with 100 mg/kg ß-glucan increased the content of intramuscular fat and changed the proportion of saturated fatty acid and unsaturated fatty acid, thereby improved the flavor of meat. In conclusion, the basal diet supplemented with 100 mg/kg Agrobacterium sp. ZX09 ß-glucan improves growth performance, nutrient digestibility, carcass length, and pork quality of finishing pigs.