Preparation of Diosgenin from Dioscorea zingiberensis： A Review / 中国实验方剂学杂志

Direct acid hydrolysis of Dioscorea zingiberensis rhizomes for preparation of diosgenin is wildly used in the traditional industry， which uses a large amount of inorganic acid catalysts， with high wastewater discharge and serious environmental pollution. Therefore， exploring clean and efficient preparation methods and processes has become an inevitable choice to realize the sustainable development of industrial production of diosgenin. Herein， the author reviewed and analyzed the research progress and problems of enzymatic hydrolysis， microbial transformation and modified acid hydrolysis in the preparation of diosgenin from D. zingiberensis rhizomes during the last ten years， and their application prospects are analyzed. Enzymatic hydrolysis has mild reaction conditions， but the yield of diosgenin is low， the economic cost is high， and the purification process of active enzyme is complicated. Microorganism shows specific activity to the substrate and high efficiency for diosgenin production， and microbial transformation is clean and environmentally friendly， but microbial transformation is time-consuming and the metabolic intermediates are complicated. For the modified acid hydrolysis， two-phase acid hydrolysis can reduce the amount of acid catalyst， and sulfonic acid-functionalized ionic liquid displays good recyclable performance by replacing the traditional inorganic acid， however， the wastewater discharge should still be considered. Solid acid catalysts are non-corrosive and easy to be recycled， but the need to use ethanol as the reaction solvent has certain safety hazards， and the catalyst preparation process is cumbersome. In conclusion， exploring clean and efficient conversion methods is an important research trend for preparation of diosgenin from D. zingiberensis rhizomes. For the enzymatic hydrolysis， the key glycoside hydrolases in the bioconversion process should be explored in depth， the conversion pathway of enzymatic saponins and enzyme specificity should be fully elucidated， and efforts should be made to improve the efficiency of enzymatic hydrolysis. For the microbial transformation， we should accelerate its industrial application process based on selecting and breeding efficient transformation strains， and optimizing stable transformation systems and processes， and in-depth investigation of the mechanism of microbial transformation， fully elucidating the specific key hydrolases and its catalytic properties， and striving to improve the efficiency of microbial transformation. For the modified acid hydrolysis， novel acid catalytic system with simple structure， stable performance and good biodegradability should be explored and applied， which can effectively solve the problems of environmental pollution and production safety.

Two new terpene glycosides from the Alpiniae Oxyphyllae Fructus / 药学学报

Two undescribed terpene glycosides and two compounds were isolated from the n-butanol fraction of Alpiniae Oxyphyllae Fructus by using various chromatographic methods, including MCI Gel, Sephadex LH-20, ODS, silica gel and semi-preparative HPLC. The structures of the isolated compounds were identified by spectroscopy methods (1D, 2D NMR, UV, IR, MS, etc.), and the absolute configuration of the compound 1 was determined by ECD calculation and acid hydrolysis. Compounds 1 and 2 are new compound, and compounds 3 and 4 were isolated from Alpiniae Oxyphyllae Fructus for the first time.

Five new terpenoids from Viburnum odoratissimum var. sessiliflorum / 中国天然药物

Five new terpenoids, including two vibsane-type diterpenoids (1, 2) and three iridoid allosides (3-5), together with eight known ones, were isolated from the leaves and twigs of Viburnum odoratissimum var.sessiliflorum. Their planar structures and relative configurations were determined by spectroscopic methods, especially 2D NMR techniques. The sugar moieties of the iridoids were confirmed as β-D-allose by GC analysis after acid hydrolysis and acetylation. The absolute configurations of neovibsanin Q (1) and dehydrovibsanol B (2) were determined by quantum chemical calculation of their theoretical electronic circular dichroism (ECD) spectra and Rh2(OCOCF3)4-induced ECD analysis. The anti-inflammatory activities of compounds 1, 3, 4, and 5 were evaluated using an LPS-induced RAW264.7 cell model. Compounds 3suppressed the release of NO in a dose-dependent manner, with an IC50 value of 55.64 μmol·L-1. The cytotoxicities of compounds 1-5 on HCT-116 cells were assessed and the results showed that compounds 2 and 3 exhibited moderate inhibitory activities with IC50 values of 13.8 and 12.3 μmol·L-1, respectively.

Comparative amino acid composition and quality parameters of Moringa oleifera testa and cotyledon

@#Introduction: Moringa oleifera is a drought-resistant plant, widely used in the tropical region. The leaves and stems have been extensively utilised in foods and neutraceuticals preparation, with less attention to the seeds. In this study, amino acid (AA) compositions of M. oleifera testa and cotyledon were examined comparatively. Methods: Samples were separately defatted, hydrolyed, and neutralised. The AA solution was purified by cation-exchange solid-phase extraction, derivatised and analysed by gas chromatography. Results: Glutamic (acidic amino acid) and phenylalanine (essential amino acid, EAA) were the most concentrated in both samples. Total EAA (g/100g crude protein, cp) was higher in cotyledon (51.0) than testa (41.9). Predicted protein efficiency ratios (P-PERs) were higher in testa (0.605-1.530) than cotyledon 0.286-1.460). EAA index ranged between 0.951-1.13 (soybean comparison) and 83.0-96.9 (egg comparison) with corresponding biological value of 78.7-93.9. The following AA had scores >1.0 in comparison to whole hen’s egg, testa: glycine (Gly), glutamic acid (Glu), phenylalanine (Phe), histidine (His), and cysteine (Cys); cotyledon (Gly), proline (Pro), Glu, Phe, His, arginine (Arg) and Cys. In comparison with requirements of pre-school children, six AA (6/9 or 66.7%) had scores >1.0 in each sample. In provisional AA scoring pattern, isoleucine (Ile) (1.25) and Phe + tyrosine (Tyr) (1.68) had scores >1.0 in testa while methionine (Met) + Cys, Phe+Tyr, and tryptophan (Trp) in cotyledon. However, tryptophan and lysine were the limiting AAs in testa and cotyledon, respectively. Conclusion: The study showed that both anatomical parts would complement each other in terms of amino acid supply.

Structure and identification of immune fragments of acid-degraded oligosaccharides from active Astragalus polysaccharides APS-Ⅱ / 药学学报

We previously reported that active Astragalus polysaccharides APS-Ⅱ generate strong immune activity. Here we establish the optimal method for APS-II acid degradation. After preliminary structural studies and separation and preparation of the degradation products, the oligosaccharide active center with the strongest immune activity was identified by in vitro immune cell culture experiments. The optimum acid degradation conditions for APS-II were determined by a single factor experiment and an orthogonal experiment. Astragalus oligosaccharides prepared under the optimal conditions were subjected to structural analysis by hydrophilic interaction chromatography - electrospray ionization source - high resolution time-of-flight mass spectrometry. The products were separated and oligosaccharide fragments with different degrees of polymerization were isolated by preparative purification chromatography. Finally, fragments of the immunologically active centers were identified by in vitro immune cell cultures from multiple perspectives. The results show that the optimal acid hydrolysis conditions for APS-Ⅱ are hydrolysis temperature 80 ℃, trifluoroacetic acid concentration 1.0 mol·L-1, hydrolysis time 1 h. The degradation conditions have good repeatability. The degradation product is a six-carbon aldehyde glycan structure with the main chain 1→4 connected. The immune activity screening experiment for six oligosaccharide fragments showed that larger molecular weight oligosaccharides have stronger immune-promoting effects. It is speculated that the immunologically active center of Astragalus oligosaccharide is located in the sugar chain of DP9-DP19. The animal welfare and the experimental process in this study follow the requirements of the Animal Ethics Committee of Shanxi University. This result suggests a foundation for the structural characterization and structure-activity relationship research of Astragalus oligosaccharides, and may promote the development of Astragalus oligosaccharide drugs.

Comparison of methodologies for fat determination and evaluation of calcium and phosphorus content in snacks for dogs / Comparação de metodologias para determinação de gordura e avaliação do conteúdo de cálcio e fósforo em petiscos para cães

In recent years, the relationship between humans and companion animals has tightened considerably and resulted in the expansion of the range of pet food industry products available in the market. In this context, snacks have gained greater popularity as pet owners seek to please their animals by providing such foods. Due to the growing importance of the snack segment, a need exists for accurate information on the nutritional composition of these products, such as fat concentration. No studies were found that evaluated the effectiveness of different methods applied for determining the content of this nutrient in dog snacks. In addition, too much mineral content can pose health risks. Thus, the objective of this study was to compare three methodologies for determining fat in pet snack products. The moisture, calcium and phosphorus content of each was also determined to compare the obtained results with each value stated on their product labels. Fat determination methods evaluated were ether extract (EE), ether extract after acid hydrolysis (EEHA), and fat content obtained from Ankom XT15 analyzer (ANKOM). Twenty-four snacks produced by 17 companies were evaluated. The results of the three methodologies were compared using the Tukey test. The comparison between the results of the laboratory analysis and the values stated on the labels was performed using descriptive statistics. There was no difference between the three methods evaluated (p = 0.34) regarding fat content. Regarding the nutritional compliance of the labels, 25% (n = 6) of the snacks presented higher moisture content than the declared amount, 50% (n = 12) presented lower fat content, 25% (n = 6) lower phosphorus content and, in 50% (n = 12), the calcium content was not within the minimum and maximum range stated on the label. Therefore, due to the absence of difference between the results, any of the three fat determination methodologies could be used. Regarding compliance of labels for calcium, phosphorus and fat content, greater control over the nutritional composition of these foods is required since most pet owners tend to supply large quantities of snacks to dogs, leading to excessive daily energy intake.(AU)

Nos últimos anos a relação entre seres humanos e animais de companhia estreitou-se consideravelmente e houve uma expansão da gama de produtos da indústria pet food disponíveis no mercado. Nesse contexto, os petiscos ganharam maior popularidade, uma vez que os tutores buscam agradar seus pets com esse tipo de alimento. Devido à crescente importância do segmento de petiscos, há a necessidade de informações precisas sobre a composição nutricional desses produtos, como o teor de gordura, uma vez que não foram encontrados estudos que avaliassem a eficácia dos métodos de determinação do teor deste nutriente em petiscos destinados a cães e o excesso de minerais pode implicar em riscos para a saúde. Assim, o presente trabalho comparou três metodologias para determinação de gordura em petiscos para cães, e também determinou os seus respectivos teores de umidade, cálcio e fósforo, cujos resultados foram comparados aos valores declarados pelos fabricantes nos rótulos dos produtos. Os métodos de determinação da gordura avaliados foram: extrato etéreo (EE), extrato etéreo após hidrólise ácida (EEHA) e teor de gordura obtido em analisador Ankom XT15 (ANKOM). Vinte e quatro petiscos produzidos por 17 empresas foram avaliados. Os resultados das três metodologias de determinação da gordura foram comparados com o emprego do teste Tukey. A comparação entre os resultados das análises laboratoriais e os valores declarados nos rótulos foi realizada por meio de estatística descritiva. Não houve diferença entre os três métodos avaliados (p = 0,34) em relação ao teor de gordura dos petiscos examinados. Em relação à conformidade nutricional dos rótulos, 25% (n = 6) dos petiscos apresentaram teor de umidade superior ao declarado, 50% (n = 12) apresentaram menor teor de gordura; 25% (n = 6) menor teor de fósforo e, em 50% (n = 12) deles, o teor de cálcio estava fora da faixa mínima e máxima declarada no rótulo. Portanto, devido à ausência de diferença entre os resultados, as três metodologias de determinação de gordura podem ser utilizadas. Quanto à conformidade dos rótulos em relação aos teores de cálcio, fósforo e gordura, é necessário maior controle sobre a composição nutricional desses alimentos, uma vez que a maioria dos tutores fornece petiscos em elevadas quantidades para os cães, que podem determinar excessivo consumo de energia.(AU)

Study on chemical constituents in acid hydrolysates of Panax notoginseng saponins / 中草药

Objective: To study the chemical constituents in acid hydrolysates of Panax notoginseng saponins (PNS). Methods: These compounds were separated and purified by column chromatography, and their structures were elucidated based on spectroscopic analyses (HR-ESI-MS, ESI-MS, 1H-NMR, 13C-NMR, HSQC and HMBC). Results: Eighteen compounds were obtained from the acid hydrolysates of PNS and characterized as dammar-25-ene-24-hydroperoxyl-3β,6α,12β,20S-tetraol (1), 6α,12β,20S-trihydroxy- dammarane-24-ene-3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (2), 6α,12β,20R-trihydroxy-dammarane-24-ene-3-O-β-D- glucopyranosyl-(1→2)-β-D-glucopyranoside (3), vina-ginsenoside-R8 (4), 24(S)-pseudo-ginsenoside-GQ (5), ginsenoside Rg5 (6), 20 (R)-ginsenoside Rg3 (7), 20(R)-ginsenoside Rk2 (8), 3β,12β-dihydroxy-dammar-(E)-20(22),24-diene-6-O-β-D-xylopyranosyl- (1→2)-β-D-glucopyranoside (9), 20(S)-ginsenoside Rg2 (10), ginsenoside SL1 (11), 20(R)-ginsenoside Rh1 (12), 20(22) E-ginsenoside Rh4 (13), 25-hydroxy-20(R) ginsenoside-Rh1 (14),3β,6α,12β,20(S)-20,25-epoxy-3,12-dihydroxy-dammarane-6-O-β-D-glucopyranoside (15), 20(R)-protopanaxadiol (16), 20(R)-protopanaxatriol (17), and 20(S)-protopanaxatriol (18). Conclusion: Compound 1 is a new triterpen saponin, and compounds 2-5 are isolated from P. notoginseng and acid dydrolysates of PNS for the first time.

Comparative study on different areas of Astragali Radix based on oligosaccharides characteristic map and immunological activity evaluation of partial acid hydrolyzed / 中草药

Objective: The characteristic maps and immunological activities of some acid hydrolysates of polysaccharides from Astragali Radix (APS) from different areas were compared. The quality evaluation method of Astragali Radix with oligosaccharide mixture as quality control index was established. Methods: In this study, the polysaccharides from the traditional Chinese medicine Astragali Radix were used as the research object. Firstly, the optimal partial acid hydrolysis conditions were selected by orthogonal test. The polysaccharide was hydrolyzed into oligosaccharides for analysis. The characteristic map of Astragalus oligosaccharides based on partial acid hydrolysis-hydrophilic interaction chromatography was established. Multivariate statistical analysis was performed on the data by SIMCA software to distinguish Mongolian Astragalus from different areas. The partial acid hydrolysis products of Astragalus polysaccharides were characterized by hydrophilic interaction chromatography and mass spectrometry, and the activity was evaluated by mouse peritoneal macrophage phagocytosis neutral red experiment. Results: The optimal hydrolysis conditions obtained by orthogonal experiment were temperature 90 ℃, trifluoroacetic acid concentration 1 mol/L, hydrolysis time 1 h. Under this condition, Astragalus polysaccharide was hydrolyzed into characteristic oligosaccharide fragments, and the method is reproducible. The characteristic map of Astragalus oligosaccharides based on partial acid hydrolysis-hydrophilic interaction chromatography showed that the maps of the same Astragalus oligosaccharides had good consistency, and the maps of different Astragalus oligosaccharides were quite different. PCA showed that three different kinds of Mongolian Astragalus can be distinguished. It was found by mass spectrometry that the extracted Astragalus polysaccharides were mainly 1→4 linear glucan, and gluco-oligosac-charides with the degrees of polymerization 3—8 were obtained after partial acid hydrolysis. The partial acid hydrolysate of the wild Astragalus polysaccharides from Shanxi Hunyuan had higher ability to enhance the phagocytic activity of macrophages than the transplanted Astragalus and higher than the unhydrolyzed total astragalus polysaccharide. Conclusion: This study showed that the characteristics of Astragalus polysaccharides based on partial acid hydrolysis-hydrophilic interaction chromatography and the effects on cellular immune function can be used to evaluate the quality of Astragali Radix in different habitats and different planting methods, and it is also an important supplement to the quality evaluation method of Astragali Radix. At the same time, it has a certain exemplary role in the characterization of other Chinese materia medica polysaccharides.

Chemical constituents from acid hydrolyzates of Panax quinquefolius total saponins and their inhibition activity to α-glycosidase and protein tyrosine phosphatase 1B / 中草药·英文版

Objective: To investigate the hypoglycemic components from the acid hydrolyzates of Panax quinquefolius total saponins, and screen the active compounds by in vitro inhibitory activities to α-glycosidase enzymes and protein tyrosine phosphatase-1B (PTP1B). Methods: The hydrolyzates were chromatographed repeatedly over silica gel column, and the structures of the compounds were determined by means of NMR. The in vitro bioassay was performed through the inhibitory effects on α-glucosidase or/and PTP1B. Results: Eight compounds were isolated, which identified as 20(S)-panaxadiol (1), (20S,24R)-dammarane-20,24-epoxy-3β,6α,12β,25-tetraol (2), 20(R)-dammarane-3β,12β,20,25-tetraol (3), 20(S)-dammarane-3β,6α,12β,20,25-pentol (4), 20(R)-dammarane-3β,12β,20,25-tetrahydroxy-3β-O-β-D-glucopyranoside (5), β-sitosterol (6), oleanolic acid (7) and 20(S)-protopanaxadiol (8). Compound 5 was ginseng triterpenoid isolated from the acid hydrolysates of total saponins from P. quinquefolius for the first time. In this paper, the possible in vitro inhibitory activities were investigated. Compound 5 exhibited significantly inhibitory activity against α-glucosidase, and the IC50 value [(0.22 ± 0.21) µmol/L] was about 43-fold lower than positive control. For the PTP1B inhibition assay, compound 5 indicated the strongest inhibitory effect with IC50 of (5.91 ± 0.38) µmol/L, followed by compound 4 with IC50 of (6.21 ± 0.21) µmol/L, which were all showed competitive inhibitory pattern by using a Lineweaver-Burk plot. Conclusion: These results supported the potential application of dammaranes from acid hydrolyzates of P. quinquefolius total saponins can be used as ingredients of ancillary anti-diabetic agent or functional factor.

Simultaneous Determination of Nine Trace D-Amino Acids in Thymalfasin by Deuterated Acid Hydrolysis-Precolumn Derivatization-Ultra-High Performance Liquid Chromatography-Quadrupole-Time of Flight Mass Spectrometry / 中国药学杂志

OBJECTIVE: To develop an ultra-high performance liquid chromatography coupled with quadrupole tandem time-of-flight mass spectrometry method for simultaneous determination of nine trace D-amino acids in thymalfasin, combined with deuterated acid hydrolysis and precolumn-derivatization. METHODS: The sample was hydrolyzed by deuterated acid, followed by precolumn-derivatization with Nα-(2, 4-dinitro-5-fluorophenyl)-L-alaninamide (FDAA). The analysis was then performed on an ACCQ-TAGTM ULTRA C18 column (2.1 mm×100 mm, 1.7 μm), with mobile phase comprising water containing 0.1% formic acid-acetonitrile gradiently eluted at a flow rate of 0.3 mL•min-1. Nine D-amino acids in thymalfasin were correctly quantified using standard curves by Xevo G2-S Q-TOF coupled with electrospray ion source in positive ion mode. RESULTS: The standard curves of the nine D-amino acids derivatives had good linearity in the ranges of the tested concentrations (r>0.991 2). The limits of quantitation of all D-amino acids derivatives were as low as 0.05-0.30 pmol. The precision and recoveries met the requirements of Chinese Pharmacopoeia (2015). The contents of D-amino acids in the samples from five companies were determined to be 25.16-1 322.01 μg•g-1. Among them, D-glutamate, D-aspartate, D-lysine and D-serine had higher contents, which were 1 105.45-1 322.01, 614.15-740.50, 358.06-388.76 and 138.78-291.60 μg•g-1, respectively. CONCLUSION: The method is sensitive, efficient and reliable, working well for simultaneous determination of nine trace D-amino acids in thymalfasin, which provides a reference for the comprehensive control of racemic peptide impurities in this synthetic polypeptide drug.

Study on Content Determination of Ellagic Acid in Different Medicinal Parts of Rosa roxburghii and in vitro Antioxidant Activity of Its Ethanol Extract / 中国药房

OBJECTIVE： Compare the contents of free ellagic acid and total ellagic acid in fruits， roots and levels of Rosa roxburghii， and to evaluate the in vitro anti-oxidant activity of ethanol extract of three medicinal parts of R. roxburghii. METHODS： Free ellagic acid and total ellagic acid were obtained from different medicinal parts of R. roxburghii by ultrasonic extraction and acid hydrolysis， respectively. The contents of them were determined by UPLC. Using half-clearance value （IC50） as anti-oxidant evaluation index， free radical scavenging test of 1，1-diphenyl-2-trinitrophenylhydrazine （DPPH） and 2，2′ -binitro-bis（3-ethylbenzothiazoline-6-sulfonic acid） （ABTS） were conducted for ethanol extract of different medicinal part， using vitamin C （VC） as positive control. RESULTS： There were significant differences in the content of free ellagic acid and total ellagic acid in different medicinal parts of R. roxburghii. The content level of free ellagic acid was in descending order： R. roxburghii leaves （38.49 mg/g）＞R. roxburghii fruits （20.59 mg/g）＞R. roxburghii roots （11.35 mg/g）； the content level of total ellagic acid was in descending order： R. roxburghii leaves （197.08 mg/g） ＞ R. roxburghii fruits （49.36 mg/g） ＞ R. roxburghii roots （21.86 mg/g）. The contents of total ellagic acid in the fruits and roots of R. roxburghii were twice as much as that of free ellagic acid in corresponding parts； the contnets of total ellagic acid in the leaves of R. roxburghii were five times higher than that of free ellagic acid in corresponding parts. In the DPPH free radical scavenging test and ABTS free radical scavenging test， the order of antioxidant activity was VC ＞ R. roxburghii leaves＞R. roxburghii fruits＞R. roxburghii roots. There was no statistical significance in the effects of R. roxburghii leaves [IC50 to DPPH free radical and ABTS free radical were （4.57±0.70）、（115.99±2.21） μg/mL] and R. roxburghii fruits [IC50 to DPPH free radical and ABTS free radical were （5.12±0.24）、（127.61±3.31）    μg/mL]， compared with the effects of VC [IC50 to DPPH free radical and ABTS free radical were （4.47±0.38）、（121.42±2.65）      μg/mL] （P＞0.05）. CONCLUSIONS： Among fruits， roots and leaves of R. roxburghii， the content of free （total） ellagic acid is the highest in the R. roxburghii leaves and in vitro anti-oxidant activity of its ethanol extract is the strongest.

Prickly pear cactus as a raw material for lactic acid production by Lactococcus lactis subsp. Lactis

@#Aims: In recent years, microbial conversion of renewable raw materials has become an important objective in industrialbiotechnology. Wastes from Opuntia ficus indica (OFI) can be considered as potential renewable raw materials in lacticacid production. In this study, the feasibility of lactic acid production using fruits and cladodes of OFI as carbohydratefeedstock was investigated.Methodology and results: Response surface methodology (RSM) based on central composite design (CCD) was usedto evaluate the effects of fermentation parameters for lactic acid production from OFI fruits by Lactococcus lactis subsp.lactis strain, isolated from Algerian raw camel milk. Acid hydrolysis of the OFI cladodes biomass was performed by diluteH2SO4 pretreatment. Lactic acid production from OFI fruits was analyzed using response surface methodology.Variables such as inoculum age and reducing sugars concentration were found to significantly influence lactic acidproduction. Final lactic acid concentration and productivity attained under optimum fermentation conditions were 32.5g/L and 0.74 g/L.h, respectively. The cladodes of OFI are a potential biomass feedstock for lactic acid production. Themaximum lactic acid and volumetric productivity were 16.85 g/L and 0.65 g/L.h, respectively.Conclusion, significance and impact of study: Wastes from OFI can be a good feedstock for lactic acid production byLactococcus lactis subsp. lactis. The methodology as a whole proved to be quite adequate for the design andoptimization of the process. The experimental results also demonstrated the feasibility of using OFI cladodeshydrolysate as a substrate for lactic acid production.

Identification of all herb medicines in Fuhuang Tablet / 中草药

Objective To establish the identification method of Typha angustifolia (stir-bake to black), Sophora japonica, Sanguisorba officinalis (stir-bake to black), and Rheum palmatum in Fuhuang tablets. Methods HPLC method were studied to identify typhaneoside and isorhamnetin-3-O-neoheptanoside in T. angustifolia, and sophoricoside in S. japonica. HPLC conditions were as follows: Agilent TC-C18 (2) column (250 mm × 4.6 mm, 5 μm) with a gradient elution at a temperature of 30 ℃; Phosphate buffer and acetonitrile were used as the mobile phase with a flow rate of 1.0 mL/min; The detection wavelength was 254 nm. The acid hydrolysis-HPLC method was explored to identify S. officinalis (stir-bake to black). TLC identification method was performed to identify R. palmatum. Results HPLC method can identify typhaneoside, isorhamnetin-3-O-neoheptanoside, and sophoricoside synchronously. Acid hydrolysis method can identify S. officinalis (stir-bake to black) by HPLC and TLC can identify R. palmatum. Conclusion The three methods are simple and accurate with high sensitivity and good specificity, which can be used to identify all herbal medicines in Fuhuang tablets.

A study on the determination of the total flavonol glycosides in health food / 预防医学

Objective To develop a HPLC method for the determination of the total flavonol glycosides in health food by acid hydrolysis. Methods The sample was extracted by methanol and was hydrolyzed into 3 aglycones by acid hydrolysis. They were quercetin, kaempferide and isorhamnetin. Chromatographic separation was carried out by the method of HPLC. The content of total flavonol glycosides was calculated. Results Methanol -25% hydrochloric acid solution (4:1) was added into selected samples, then 85 ℃ water bath heating and refluxing were conducted for 30 min. Optimum chromatographic conditions were as follows. Chromatographic column: ODS C18 (150 mm ×4.6 mm,5μm) . Mobile phase: used methanol-0.6% phosphoric acid (47:53) . Column temperature was set at 35 ℃. Detection wavelength was 360 nm. The linear ranges of quercetin, kaempferide and isorhamnetin were (4.766-95.52) μg/mL(r=0.9991) , (5.052-101.0) μg/mL (r=0.9994) and (2.027-48.66) μg/mL (r=0.9994) respectively. The relative standard deviation of the samples was 0.57%. The recovery rate was 98.25% (RSD=3.82%), 99.81% (RSD=3.31%) and 101.8% (RSD=4. 86%) . The detection limits of quercetin and kaempferide were both 0.50 μg, while Isorhamnetin was 0.80 μg, the total flavonol glycosides was 4.52 μg. The content of total flavonol glycosides detected by this method was lower than the content of the total glavonoids detected by the method provided by technical specification for inspection and evaluation of health food (2003 Edition) . Conclusion This method proved to be simple, stable and had strong repeatability and could be used for the determination of total flavonol glycosides in health food.

Structural characterization of Glehniae Radix polysaccharides using partial acid hydrolysis-hydrophilic interaction liquid chromatography-mass spectrometry / 中国中药杂志

Water-soluble polysaccharides from traditional Chinese medicine have properties of complex structure and high molecular, resulting in hardly complete their structural characterization.However, a "bottom-up" approach could solve this problem.Glehniae Radix extract was extracted with hot water and then precipitated by 40% ethanol to obtain Glehniae Radix polysaccharides (RGP). Subsequently, a partial acid hydrolysis method was carried out and the effects of acid concentration, time and temperature on hydrolysis were investigated. Under the optimum hydrolysis condition (1.5 mol•L⁻¹ trifluoroacetic acid, 4 h, and 80 ℃), RGP were hydrolyzed to characteristic oligosaccharide fragments. Futher, a hydrophilic liquid chromatography- mass spectrometry method was used for the separation and structural characterization of the polysaccharide hydrolysates. According to MS and MS/MS analysis of several standard disaccharides, a method for determining the type of polysaccharide glycosidic linkage by mass spectrometry was established. The results showed that the polysaccharide hydrolysates were linear glucan containing 1, 4-glycosidic bonds. And gluco-oligosaccharides with the degrees of polymerization (DP) of 4-11 were obtained after partial acid hydrolysis.

Nutritional composition and evaluation of different methodologies for fat determination in wet feed for dogs and cats / Composição nutricional e avaliação de diferentes métodos de determinação de gordura em alimentos úmidos para cães e gatos

As a consequence of the increasing number of dog and cat owners, the pet food industry is expanding the range of pet food products in the market. In order to obtain more necessary information about the wet food segment for dogs and cats, the aim of this study was to determine the nutritional composition, to evaluate the information declared on the labels, and to compare the composition with the FEDIAF recommendations for protein and fat. Furthermore, three different methodologies of fat analysis were compared: crude fat (CFa), crude fat after acid hydrolysis (CFAH), and fat content obtained with Ankom XT15 (ANKOM) to determine the most adequate method for fat determination in wet foods. Twenty-five wet food products were evaluated, 13 wet foods for dogs and 12 for cats. Centesimal composition analyses obtained in this study were compared with guaranteed analysis declared on the label and with FEDIAF minimum recommended requirements for each species. The results of the nutritional composition and the values described on the label and the evaluation of the three fat determination methods were compared using the mixed model test with repeated measurements in the same samples, respectively (p < 0.05) in the SAS program, evaluation of protein adequacy and fat content were analyzed by mathematical calculations of difference and proportion. No difference was observed between nutritional composition of wet foods and the values declared on the labels for the majority of the diets analyzed, and there was a predominance of products that exceeded FEDIAF minimum recommendations of protein and fat for both species. No difference was observed between the three methods of fat content evaluation (p = 0.68). It was concluded that wet foods evaluated in this study match the label information and FEDIAF nutrient requirement recommendations, considering recommended calorie intake. All three fat determination methodologies evaluated were similar, justifying the choice of the easiest or cheapest method.(AU)

Devido ao aumento do número de cães e gatos domiciliados, a indústria de alimentos para animais de estimação tem expandido a gama de produtos existentes no mercado de pet food. Para obter informações mais relevantes sobre o segmento de alimentos úmidos para cães e gatos, este trabalho determinou a composição nutricional, avaliou as informações declaradas nos rótulos e comparou a composição com as recomendações da Fediaf de proteína e gordura. Também foram comparadas três metodologias diferentes de análise de gordura: extrato etéreo (CFa), extrato etéreo após hidrólise ácida (CFAH) e teor de gordura obtido no analisador Ankom XT15 (ANKOM) para determinar o método mais adequado de avaliação de gordura em alimentos úmidos. Foram avaliadas 25 marcas de alimentos úmidos, 13 para cães e 12 para gatos. As análises de composição centesimal obtidas neste estudo foram comparadas com a informação nutricional declarada nos rótulos e com as necessidades mínimas recomendadas pela Fediaf para cada espécie. Os resultados da composição nutricional, os valores descritos no rótulo e a avaliação dos três métodos para determinação da gordura foram comparados com o emprego do teste t e modelo misto com medidas repetidas nas mesmas amostras, respectivamente (p < 0,05) no programa SAS. Já a avaliação da adequação nutricional de proteína e do teor de gordura foram analisados por cálculos matemáticos de diferença e proporção. Para a maioria dos alimentos avaliados não foi observada diferença entre a composição nutricional dos alimentos úmidos e os valores declarados em rótulo, e houve predominância de produtos que excederam as recomendações mínimas de proteína e gordura da Fediaf para ambas as espécies. Quanto às metodologias de extração de gordura, não foi observada diferença entre os três métodos avaliados (p = 0,68). Concluiu-se que os alimentos úmidos avaliados atendem às informações declaradas pelos fabricantes e também às recomendações nutricionais da Fediaf com base na ingestão energética recomendada. Em relação às metodologias avaliadas para determinação de gordura nestes alimentos, a similaridade entre tais resultados justifica o uso da técnica de maior facilidade ou de menor custo.(AU)

PRODUCCIÓN DE XILITOL POR Candida guilliermondii A PARTIR DE FERMENTACIÓN DE RESIDUOS DE PALMA DE ACEITE

La hidrólisis ácida diluida del residuo lignocelulósico de raquis de palma de aceite produce azúcares fermentables, como la xilosa, principal fuente de carbono para la producción de xilitol, por Candida guilliermondii En este estudio, se evaluó el efecto de diferentes medios de cultivo y de condiciones de fermentación sobre la producción de xilitol, a partir de raquis de palma de aceite, utilizando C guilliermondii. El hidrolizado ácido de raquis de palma suplementado con 4g/L extracto de levadura, 3g(NH4)2SO4/L, 0,5g/ MgSO4.7H2O/L y 0,1gCaCl2.2H2O/L mostró ser el mejor medio para el crecimiento de la levadura en cultivo sumergido, debido a que presentó los valores mayores, estadísticamente significativos, de la velocidad específica máxima de crecimiento, de 0,12h-1 y producción de biomasa, de 5,5g/L (p<0,05). Las condiciones de fermentación más apropiadas se obtuvieron con hidrolizado de raquis suplementado a pH de 5,5, concentración inicial de xilosa de 17g/L y un inóculo de 3g/L. La microaerobiosis mostró ser un factor importante en el proceso de fermentación, con un volumen de 40mL de medio, en un matraz de 100mL, se produjo la mayor concentración de xilitol de 6,7 g/L (p<0,05).

The dilute-acid hydrolysis of oil palm empty fruit bunch produces fermentable sugars such as xylose, main carbon source for xylitol production by Candida guilliermondii. The influence of different culture media and fermentation conditions were evaluated on the production of xylitol from oil palm empty fruit bunch using C guilliermondii. The acid hydrolyzated oil palm empty fruit bunch supplemented with4g yeast extract/L, 3g (NH4)2SO4/L, 0.5g MgSO4.7H2O/L y 0.1g CaCl2.2H2O/L, was the best culture medium for the yeast growth in submerged culture because it had the highest values, statistically significant, of the specific growth rate 0.12h-1 and biomass production 5.5g/L (p<0,05). The suitable fermentation conditions were obtained with hydrolyzated oil palm empty fruit bunch supplemented at pH 5.5, initial xylose concentration of 17g/L and an inoculum of 3g/L. Microaerobic condition is an important factor on the fermentation process, with a volume of 40mL in a flask of 100mL, that produces the highest concentrations of xylitol (6.7g/L) (p<0,05).

Influencia del tamaño de partícula y la velocidad de agitación sobre el rendimiento de pectina / Influence of the particle size and the stirring speed over the pectin yield

Introducción: el procesamiento de las hojas de sábila (Aloe barbadensis Mill) para la obtención de productos de interés agroalimentario como el gel, genera varios residuos, entre ellos el bagazo que se obtiene de la molienda de la pulpa. El mismo constituye un recurso económico para la obtención de componentes de amplio uso en las industrias alimentaria y farmacéutica, como la pectina, compuesto de interés por su alto poder gelificante, cuyo proceso de extracción más usado a nivel industrial es la hidrólisis ácida. La extracción de pectina a partir del bagazo de sábila se estudió a escala de laboratorio y se establecieron las condiciones de operación (temperatura, pH y tiempo de extracción) requeridas para maximizar el rendimiento de la pectina, sin embargo, falta estudiar la influencia de la velocidad de agitación y el tamaño de partícula. Objetivo: evaluar la influencia del tamaño de la partícula y la velocidad de agitación en el rendimiento de extracción de la pectina al utilizar la hidrólisis ácida. Métodos: se aplicó la técnica de hidrolisis ácida a escala de laboratorio para extraer pectina del bagazo de sábila. Los factores estudiados fueron: velocidad de agitación (300 y 1 000 min-1) y tamaño de partícula (0,250 y 0,600 mm). El resto de los parámetros se mantuvieron constante: tiempo de reacción (60 minutos), temperatura (90 °C), pH (1,5). La relación soluto/solvente fue fijada en 1:15 y 1:20 m/v, teniendo en cuenta el desarrollo experimental de cada tamaño de partícula. Resultados: las condiciones óptimas en el rango estudiado fueron velocidad de agitación de 1 000 min-1 y tamaño de partícula 0,600 mm y se obtiene un rendimiento de 3,80 por ciento de pectina por cada 5 g de muestra de bagazo seco. Conclusiones: es la velocidad de agitación la que ejerce una mayor influencia, aunque ambos factores resultaron significativos con un 95 por ciento de confiabilidad(AU)

Introduction: the processing of sabila leaves (Aloe barbadensis Mill) to obtain products of agricultural interest such as gel, generates various wastes, including bagasse obtained by grinding the pulp. Bagasse is an economic resource for the production of components widely used in the food and pharmaceutical industries like pectin, an interesting compound because of its high gellifying power and the most used extraction process at industrial level is acid hydrolysis. In particular, the pectin extraction from sabila bagasse was studied on a lab scale; and the operating conditions (temperature, pH and extraction time) required to maximize the pectin yield was established. However, the influence of the stirring speed and the particle size remained to be studied. Objective: to assess the influence of the particle size and the stirring speed on the extraction yield of pectin using acid hydrolysis. Methods: the acid hydrolysis technique was applied to extract pectin from the sabila bagasse. The studied factors were stirring rate (300 to 1 000 min-1) and particle size (0,250 to 0,600 mm). The rest of parameters remained unchanged: reaction time (60 minutes), temperature (90 °C) and pH (1.5). The solute/solvent ratio was set at 1:15 and 1:20 w/v, considering experimental behavior of each particle size. Results: the optimal conditions in the studied range were stirring rate 1 000 min-1 and particle size of 0,600 mm, reaching a pectin yield of 3,80 percent per 5 g sample o dry bagasse. Conclusions: the stirring rate the most influential although both factors were significant with 95 percent reliability(AU)

Process optimization of acid hydrolysis of burnet total saponins for preparation of burnet sapogenin / 中草药

Objective: To optimize the acid hydrolysis process of burnet (Sanguisorbae Radix, the roots of Sanguisorba officinalis) total saponins and to improve the yield of the hydrolyzate burnet sapogenin. Methods: Using orthogonal test design, the concentration of hydrochloric acid, solid-liquid ratio, hydrolysis temperature, and hydrolytic time for the hydrolysis process of burnet total saponins were investigated. Results: The primary and secondary factors that influenced the hydrolysis were followed by order of the concentration of hydrochloric acid > hydrolytic time > solid-liquid ratio > hydrolytic temperature, preferably optimum for the solid-liquid ratio of 1:200, hydrochloric acid concentration of 4 mol/L, hydrolytic time of 0.5 h, and hydrolytic temperature of 92 ℃. Conclusion: High yield of burnet sapogenin can be obtained by using the optimized hydrolytic conditions, which are suitable for industrial production.

Isolation and idetification of a new aglycone from acid hydrolysis products of saponin in Centella asiatica / 中草药

Objective: To study the chemical constituents of the acid hydrolysis products, in order to find some new aglycones from the hydrolysate of the saponin of Centella asiatica. Methods: The acid hydrolytic product was separated by repeated silica gel, Sephadex LH-20 chromatography. The cytotoxic activities in vitro of some compounds isolated were evaluated. Their structures isolated were identified by analysis of their spectral data of NMR, MS, IR, and UV. Results: Four aglycones, 2α,3β,23-trihydroxyurs-6,12-dien-28-oic acid (1), asiatic acid (2), 6β-hydroxy asiatic acid (3), terminolic acid (4), were isolated from the hydrolysis product. Conclusions: Compound 1, named centella-6-ene, was a new ursane-type aglycone and it showed no cytotoxic activities against the tested cancer cell lines.