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To optimize the formulation and technology of oxymatrine-astragaloside IV coloaded liposomes (Om-As-Lip) based on quality by design (QbD) principles, and further to verify the feasibility of its amplification process, Om-As-Lip was prepared by ethanol injection combined with pH gradient method. The critical material attributions of Om-As-Lip were evaluated by dual-risk analysis tools and Plackett-Burman design (PBD). The formulation of Om-As-Lip was further optimized with the Box-Behnken design (BBD). The design space was also established based on the contour plots of BBD. In order to further investigate the amplification process of Om-As-Lip, the critical process parameters of high-pressure homogenization (HPH) were optimized by single-factor test, and the quality of the final product was also evaluated. The results of risk analysis and PBD confirmed that the astragaloside concentration, cholesterol concentration, and phospholipid ratio (HSPC∶SPC) were the ctitical material attributes. The model established by BBD had a good predictability, and the optimized mass ratio of As to phospholipids was 1∶40, cholesterol to phospholipids was 1∶10, HSPC to SPC was 51∶9. The design space of Om-As-Lip was as follows: the ratio of cholesterol to phospholipids was 1∶12-1∶5 and HSPC to SPC was 1∶7-17∶3. The optimized high-pressure homogenization pressure was 600 bar, temperature was 4 ℃, and cycle times was 6 times for HPH-Om-As-Lip. The quality of Om-As-Lip prepared based on the QbD concept can meet the expected CQAs, and the formulation and technology established can provide a reliable experimental basis for its future development and applications.
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Objective:To establish reverse-phase high-performance liquid chromatography and simultaneously determine gallic acid, methyl gallate, corilagin, Sennoside B, and Sennoside A levels in Sana preparations.Methods:From January to December 2021, Phenomenex Hydro-RP 80A column (4.60 mm × 250 mm, 4 μm) was used. Elution was conducted using mobile phases methanol (A)-0.2% formic acid (B). The following gradients were applied: 1%-3%A for 0-18 minutes, 3%-15%A for 18-19 minutes, 15%-17%A for 19-40 minutes, 17%-25%A for 40-45 minutes, 25%-35%A for 45-65 minutes, 35%-60%A for 65-95 minutes, 60%-90%A for 95-96 minutes, 90%-1%A for 96-97 minutes. The flow rate was 1.0 mL/minute. The column temperature was 35 ℃. The detection wavelength was 270 nm.Results:The linear ranges of gallic acid, methyl gallate, corilagin, Sennoside B and Sennoside A were 0.182-1.099 μg ( r = 0.999 9), 0.046-0.278 μg ( r = 0.999 2), 0.266-1.598 μg ( r = 0.999 4), 0.172-1.036 μg ( r = 0.999 2), and 0.176-1.056 μg ( r = 0.999 9). The average dosing recovery rates were 100.02%, 99.14%, 99.38%, 101.77%, and 100.92%, respectively. Conclusion:Reverse-phase high-performance liquid chromatography can be used for quality control of Sana preparations because of high accuracy, sensitivity, reliability, and reproduction.
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Abstract Introduction: Various methods of analysis for the assay of chemotherapeutic agent 5-Fluorouracil (5-FU) in human and animal biological fluids have previously been reported. However, there is no standardization for detecting 5-FU in the hamsters' saliva that received the chemotherapeutic agent. Objective: Considering that the administration of 5-FU in some way changes the morphology and function of the salivary glands, and that the presence of the chemotherapeutic agents in the oral mucosa may lead to some oral complications, the aim of this study was to determine the presence of 5-FU in the hamsters' saliva that received the chemotherapeutic agent, by means of the High Performance Liquid Chromatography technique (HPCL) since this animal model is used in studies of 5-FU induced oral mucositis and glandular hypofunction. Methods: Twelve animals were divided into 4 groups: CP and CPI, in which the animals received pilocarpine (CP) or pilocarpine + isoproterenol (CPI) and the chemotherapy vehicle intraperitoneally; and Groups QP and QPI, in which the animals received the same secretagogues listed above, and the chemotherapeutic agent 5-FU, respectively. After the secretagogue administration, saliva was collected from all the animals for a period of 60 mins. Subsequently, the saliva was frozen at -80 ËC for later determination of the chemotherapeutic agent by HPLC. After the the chromatograms analysis, and based on the results obtained, it was possible to identify the presence of 5-FU in the saliva samples from hamsters that received the chemotherapeutic agent intraperitonally, by the HPLC technique. (AU)
Resumo Vários métodos de análise para o ensaio do quimioterápico 5-Fluorouracil (5-FU) em fluidos biológicos de humanos e animais, foram previamente relatados. No entanto, não há uma padronização para detecção de 5-FU na saliva de hamsters que receberam o quimioterápico. Considerando que a administração do 5-FU altera de alguma maneira a morfologia e função das glândulas salivares, e que a presença do quimioterápico na mucosa oral pode levar a algumas complicações orais, este trabalho teve como objetivo de determinar a presença de 5-FU na saliva de hamsters que receberam o quimioterápico pela técnica de Cromatografia Líquida de Alta Eficiência (CLAE), uma vez que este modelo animal é usado nos estudos com mucosite oral e hipofunção glandular, induzidas por 5-FU. Doze animais foram divididos em 4 grupos: CP e CPI, onde os animais receberam intraperitonealmente pilocarpina (CP) ou pilocarpina + isoproterenol (CPI) e o veículo do quimioterápico, e os grupos QP e QPI, onde os animais receberam, respectivamente, os mesmos secretagogos listados acima e o quimioterápico 5-FU. Após a administração do secretagogo, foi coletada a saliva de todos os animais, por um período de 60 min. Em seguida, a saliva foi congelada a -80 ËC para posterior determinação do quimioterápico por CLAE. Após análise dos cromatogramas, e com base nos resultados obtidos, foi possível identificar a presença do 5-FU nas amostras de saliva de hamsters que receberam o quimioterápico via intraperitoneal pela técnica da CLAE. (AU)
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Objective:To explore the relation of the radiochemical purity and in vivo imaging effect of 68Ga-1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid (DOTA)- D-phe1-Tyr3-Thr8-octreotide (TATE) injection. Methods:High performance liquid chromatography (HPLC) and thin-layer chromatography (TLC) methods were established to determine 68Ga-DOTATATE, 68Ga 3+ , 68Ga in colloidal form and 68Ga-DOTA- D-Phe1-Tyr3-Thr8-dethreonine-octreotide (heptapeptide) and to study the influence of precursor purity on radiochemical purity of labelled products. The uptake of 68Ga-DOTATATE injection with different radiochemical purities was investigated in nude mice bearing AR42J cells by microPET imaging and the tumor target/non-target (T/NT) value was calculated. One-way analysis of variance and Pearson correlation analysis were used to analyze the data. Results:The contents of 68Ga 3+ and 68Ga in colloidal form were not related with precursor purity ( r values: 0.385, 0.497, P values: 0.306, 0.137), while the content of 68Ga-DOTA-heptapeptide was positively related with the purity of DOTA-heptapeptide ( r=0.957, P<0.001). The radiochemical purities of 68Ga-DOTATATE injection were (87.0±2.3)%, (86.8±0.8)% and (94.0±3.1)% when the DOTATATE purities were 90.9%, 91.6% and 99.2%, respectively. The results of microPET imaging showed that the tumor uptake was positively related with the radiochemical purity of 68Ga-DOTATATE injection ( r=0.828, P<0.001), and the T/NT values of 68Ga-DOTATATE injection with radiochemical purities of 95.7%, 85.8%, 84.5% and 79.9% were 21.25±8.84, 8.50±1.51, 11.38±1.65 and 6.01±0.99, respectively ( F=11.48, P=0.001). Conclusion:The radiochemical purity of 68Ga-DOTATATE injection is impacted by the purity of labelled precursor and manufacturing processes and is related with the imaging effect in vivo.
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Objective:To investigate the effect of ultrasonic debridement mediated by 0.9% sodium chloride solution and 0.5% iodophor volt combined with eddy current washing and high pressure pulse washing on the removal of colonized bacteria on the wound surface of diabetic foot and wound healing.Methods:From March to November 2020, a total of 60 patients using ultrasonic therapy for debridement were divided into control group, experimental group 1, experimental group 2 and experimental group 3 by random digit table in the fourth People′s Hospital of Dalian. The final effective data collected was 15 cases in each group. The control group was given ultrasonic debridement mediated by 0.9% sodium chloride solution and eddy current washing.Experimental group 1 was given ultrasonic debridement mediated by 0.9% sodium chloride solution and high pressure pulse washing. Experimental group 2 received 0.5% iodophor mediated ultrasonic debridement and eddy current washing. Experimental group 3 0.5% iodophor mediated ultrasonic debridement and high pressure pulse washing. The size of the wound was measured, sampled and bacterial cultured before and after the first, fifth and 10th intervention. The wound bacterial clearance rate and wound area reduction rate were calculated and compared.Results:Before and after 3 interventions, the bacterial clearance rate and the total reduction of wound surface in 4 groups were increased ( P<0.01), the total bacterial clearance rate of experimental group 3 was the highest, which was (93.85 ± 9.87)%.The total reduction rate of wound in experimental group 2 was the highest, which was (20.831 4 ± 9.379 8)%. Conclusions:0.5% iodophor mediated ultrasonic debridement combined with high pressure pulse washing is the most effective way in the removal of diabetic foot wounds, and 0.5% iodophor solution mediated ultrasonic debridement combined with eddy current washing is the most effective in reducing diabetic foot wounds.
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Background Noise can cause not only auditory system injury, but also liver damage. However, the biomarkers and pathological mechanism of noise-induced liver injury are not clear yet. Objective To observe the effect of noise on the morphological structure and functions of rat liver. Methods A total of 30 Wistar rats were randomly divided into a normal control group, a low noise exposure group [(95 dB sound pressure level (SPL)], and a high noise exposure group (105 dB SPL). After 30 days of noise exposure, blood was collected, and livers were harvested and fixed. The pathological changes of livers were observed. The levels of biochemical indicators of liver function, blood glucose, and blood lipid were measured. Serum metabolites were detected by ultra-high-pressure liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS). Differential metabolite markers and metabolic pathways were identified. Results Compared with the control group, the body weight gain decreased in the low noise group and the high noise group after noise exposure (P<0.001, P<0.05). The pathological results showed that noise caused the rat livers’ morphological and structural damage at various degrees, and damage of the high noise exposure group was more serious. Compared with the control group, the serum levels of aspartate aminotransferase, albumin, and glycosylated serum protein in the low noise exposure group were increased (P<0.05), but the total bile acid level was decreased (P<0.05). The serum levels of alanine aminotransferase, aspartate aminotransferase, albumin, triglyceride, low density lipoprotein, and glycosylated serum protein in the high noise group exposure were increased (P<0.05), but the glucose level was decreased (P<0.05). In the serum metabolomics analysis, 11 differential metabolites were screened out in the low noise exposure group, which were mainly enriched in 3 pathways (thiamine metabolism, primary bile acid biosynthesis, and bile secretion) related to liver metabolism. Four differential metabolites were screened out in the high exposure noise group, which were mainly enriched in four significantly different metabolic pathways (insulin signaling pathway, non-alcoholic fatty liver disease, bile secretion, and insulin secretion). All the metabolic pathways involved in bile acid secretion and metabolism. Conclusion Nosie exposure can not only damage the liver structure of rats, but also affects the metabolism functions of liver. The mechanism may be related to bile acid secretion metabolic pathway.
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Objective:To analyze the changes in serum metabolites of patients with uremia using ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS), and provide a theoretical basis for the prevention and treatment of uremia.Methods:Uremia patients from the Department of Nephrology, the Affiliated Hospital of Southwest Medical University, and the volunteers from the Health Examination Center were enrolled in this study. According to the inclusion and exclusion criteria, 20 uremia patients (experimental group) and 20 volunteers (control group) were screened out. UHPLC-MS was used to detect the metabolites in the serum of subjects from the two groups, and difference analysis was made to screen the different metabolites, followed by correlation analysis and pathway enrichment study.Results:A total of 412 metabolites were identified by UHPLC-MS. Principal components analysis (PCA) proved that these metabolites could distinguish the control group and the experimental group well. The criteria [variable importance for the projection (VIP)>1, fold changes (FC)>1.25 or FC<0.8 and P value<0.05] was set to screen those significantly different metabolites. Finally, 28 significantly different metabolites were screened out, of which 18 metabolites increased significantly, the other 10 different metabolites decreased significantly. Correlation analysis results proved a certain correlation among 28 different metabolites and the experimental group and control group samples, and between the 28 differential metabolites themselves. Enrichment analysis found that 28 different metabolites might enrich the catecholamine biosynthetic pathway, and pathway analysis suggested that 28 different metabolites might affect glutamate, aspartame acid and glutamate metabolic pathways. Conclusion:Based on metabonomic analysis, some metabolites in the serum of patients with uremia have changed, which can affect some metabolic pathways, thus affecting the pathophysiological process of patients with uremia.
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The Brazilian Cerrado biome consists of a great variety of endemic species with several bioactive compounds, and Anadenanthera peregrina (L.) Speg is a promising species. In this study, we aimed to perform phytochemical characterization and evaluate the antioxidant and antibacterial activities against Staphylococcus aureus and Escherichia coli of the hydroethanolic extract of A. peregrina stem bark. The barks were collected in the Botanical Garden of Goiânia, Brazil. The hydroethanolic extract was obtained by percolation and subjected to physicochemical screening, total phenolic content estimation, high-performance liquid chromatography (HPLC) fingerprinting, and antioxidant (IC50 values were calculated for the 2,2-diphenyl-1-picrylhydrazyl assay - DPPH) and antibacterial activity determination. The pH of the extract was 5.21 and density was 0.956 g/cm3. The phytochemical screening indicated the presence of cardiac glycosides, organic acids, reducing sugars, hemolytic saponins, phenols, coumarins, condensed tannins, flavonoids, catechins, depsides, and depsidones derived from benzoquinones. The extract showed intense hemolytic activity. The total phenolic content was 6.40 g GAE 100 g-1. The HPLC fingerprinting analysis revealed the presence of gallic acid, catechin, and epicatechin. We confirmed the antioxidant activity of the extract. Furthermore, the extract did not inhibit the growth of E. coli colonies at any volume tested, but there were halos around S. aureus colonies at all three volumes tested. These results contribute to a better understanding of the chemical composition of A. peregrina stem bark and further support the medicinal applications of this species.
O bioma Cerrado brasileiro apresenta em uma grande variedade de espécies endêmicas com diversos compostos bioativos, e Anadenanthera peregrina (L.) Speg é uma espécie promissora. Neste estudo, objetivamos realizar a caracterização fitoquímica e avaliar as atividades antioxidantes e antibacterianas contra Staphylococcus aureus e Escherichia coli do extrato hidroetanólico de cascas do caule de A. peregrina. As cascas foram coletadas no Jardim Botânico de Goiânia, Brasil. O extrato hidroetanólico foi obtido por percolação e submetido a triagem físicoquímica, estimativa de conteúdo fenólico total, impressão digital por cromatografia líquida de alta eficiência (HPLC) e determinação da atividade antioxidante (valores de IC50 foram calculados para o ensaio 2,2-difenil-1-picril-hidrazil) e antibacteriana. O pH do extrato foi de 5,21 e a densidade foi de 0,956 g/cm3. A triagem fitoquímica indicou a presença de glicosídeos cardíacos, ácidos orgânicos, açúcares redutores, saponinas hemolíticas, fenóis, cumarinas, taninos condensados, flavonóides, catequinas, depsídios e depsidonas derivados de benzoquinonas. O extrato mostrou intensa atividade hemolítica. O conteúdo fenólico total foi de 6,40 g de GAE 100 g-1. A análise por impressão digital por HPLC revelou a presença de ácido gálico, catequina e epicatequina. Confirmamos a atividade antioxidante do extrato. Além disso, o extrato não inibiu o crescimento de colônias de E. coli em nenhum volume testado, mas houve halos em torno das colônias de S. aureus nos três volumes testados. Estes resultados contribuem para uma melhor compreensão da composição química da casca de A. peregrina e apoia ainda mais as aplicações medicinais desta espécie.
Subject(s)
Plant Bark , Antioxidants/pharmacology , Staphylococcus aureus , Brazil , Plant Extracts/pharmacology , Escherichia coli , Phytochemicals , Anti-Bacterial Agents/pharmacologyABSTRACT
The Brazilian Cerrado biome consists of a great variety of endemic species with several bioactive compounds, and Anadenanthera peregrina (L.) Speg is a promising species. In this study, we aimed to perform phytochemical characterization and evaluate the antioxidant and antibacterial activities against Staphylococcus aureus and Escherichia coli of the hydroethanolic extract of A. peregrina stem bark. The barks were collected in the Botanical Garden of Goiânia, Brazil. The hydroethanolic extract was obtained by percolation and subjected to physicochemical screening, total phenolic content estimation, high-performance liquid chromatography (HPLC) fingerprinting, and antioxidant (IC50 values were calculated for the 2,2-diphenyl-1-picrylhydrazyl assay - DPPH) and antibacterial activity determination. The pH of the extract was 5.21 and density was 0.956 g/cm3. The phytochemical screening indicated the presence of cardiac glycosides, organic acids, reducing sugars, hemolytic saponins, phenols, coumarins, condensed tannins, flavonoids, catechins, depsides, and depsidones derived from benzoquinones. The extract showed intense hemolytic activity. The total phenolic content was 6.40 g GAE 100 g-¹. The HPLC fingerprinting analysis revealed the presence of gallic acid, catechin, and epicatechin. We confirmed the antioxidant activity of the extract. Furthermore, the extract did not inhibit the growth of E. coli colonies at any volume tested, but there were halos around S. aureus colonies at all three volumes tested. These results contribute to a better understanding of the chemical composition of A. peregrina stem bark and further support the medicinal applications of this species.
O bioma Cerrado brasileiro apresenta em uma grande variedade de espécies endêmicas com diversos compostos bioativos, e Anadenanthera peregrina (L.) Speg é uma espécie promissora. Neste estudo, objetivamos realizar a caracterização fitoquímica e avaliar as atividades antioxidantes e antibacterianas contra Staphylococcus aureus e Escherichia coli do extrato hidroetanólico de cascas do caule de A. peregrina. As cascas foram coletadas no Jardim Botânico de Goiânia, Brasil. O extrato hidroetanólico foi obtido por percolação e submetido a triagem físico química, estimativa de conteúdo fenólico total, impressão digital por cromatografia líquida de alta eficiência(HPLC) e determinação da atividade antioxidante (valores de IC50 foram calculados para o ensaio 2,2-difenil-1-picril-hidrazil) e antibacteriana. O pH do extrato foi de 5,21 e a densidade foi de 0,956 g/cm3. A triagem fitoquímica indicou a presença de glicosídeos cardíacos, ácidos orgânicos, açúcares redutores, saponinas hemolíticas, fenóis, cumarinas, taninos condensados, flavonóides, catequinas, depsídios e depsidonas derivados de benzoquinonas. O extrato mostrou intensa atividade hemolítica. O conteúdo fenólico total foi de 6,40 g de GAE 100 g-1. A análise por impressão digital por HPLC revelou a presença de ácido gálico, catequina e epicatequina. Confirmamos a atividade antioxidante do extrato. Além disso, o extrato não inibiu o crescimento de colônias de E. coli em nenhum volume testado, mas houve halos em torno das colônias de S. aureus nos três volumes testados. Estes resultados contribuem para uma melhor compreensão da composição química da casca de A. peregrina e apoia ainda mais as aplicações medicinais desta espécie.
Subject(s)
Plant Extracts/analysis , Plant Extracts/pharmacology , Fabaceae , Plants, Medicinal/chemistryABSTRACT
Abstract The Brazilian Cerrado biome consists of a great variety of endemic species with several bioactive compounds, and Anadenanthera peregrina (L.) Speg is a promising species. In this study, we aimed to perform phytochemical characterization and evaluate the antioxidant and antibacterial activities against Staphylococcus aureus and Escherichia coli of the hydroethanolic extract of A. peregrina stem bark. The barks were collected in the Botanical Garden of Goiânia, Brazil. The hydroethanolic extract was obtained by percolation and subjected to physicochemical screening, total phenolic content estimation, high-performance liquid chromatography (HPLC) fingerprinting, and antioxidant (IC50 values were calculated for the 2,2-diphenyl-1-picrylhydrazyl assay - DPPH) and antibacterial activity determination. The pH of the extract was 5.21 and density was 0.956 g/cm3. The phytochemical screening indicated the presence of cardiac glycosides, organic acids, reducing sugars, hemolytic saponins, phenols, coumarins, condensed tannins, flavonoids, catechins, depsides, and depsidones derived from benzoquinones. The extract showed intense hemolytic activity. The total phenolic content was 6.40 g GAE 100 g-1. The HPLC fingerprinting analysis revealed the presence of gallic acid, catechin, and epicatechin. We confirmed the antioxidant activity of the extract. Furthermore, the extract did not inhibit the growth of E. coli colonies at any volume tested, but there were halos around S. aureus colonies at all three volumes tested. These results contribute to a better understanding of the chemical composition of A. peregrina stem bark and further support the medicinal applications of this species.
Resumo O bioma Cerrado brasileiro apresenta em uma grande variedade de espécies endêmicas com diversos compostos bioativos, e Anadenanthera peregrina (L.) Speg é uma espécie promissora. Neste estudo, objetivamos realizar a caracterização fitoquímica e avaliar as atividades antioxidantes e antibacterianas contra Staphylococcus aureus e Escherichia coli do extrato hidroetanólico de cascas do caule de A. peregrina. As cascas foram coletadas no Jardim Botânico de Goiânia, Brasil. O extrato hidroetanólico foi obtido por percolação e submetido a triagem físico-química, estimativa de conteúdo fenólico total, impressão digital por cromatografia líquida de alta eficiência (HPLC) e determinação da atividade antioxidante (valores de IC50 foram calculados para o ensaio 2,2-difenil-1-picril-hidrazil) e antibacteriana. O pH do extrato foi de 5,21 e a densidade foi de 0,956 g/cm3. A triagem fitoquímica indicou a presença de glicosídeos cardíacos, ácidos orgânicos, açúcares redutores, saponinas hemolíticas, fenóis, cumarinas, taninos condensados, flavonóides, catequinas, depsídios e depsidonas derivados de benzoquinonas. O extrato mostrou intensa atividade hemolítica. O conteúdo fenólico total foi de 6,40 g de GAE 100 g-1. A análise por impressão digital por HPLC revelou a presença de ácido gálico, catequina e epicatequina. Confirmamos a atividade antioxidante do extrato. Além disso, o extrato não inibiu o crescimento de colônias de E. coli em nenhum volume testado, mas houve halos em torno das colônias de S. aureus nos três volumes testados. Estes resultados contribuem para uma melhor compreensão da composição química da casca de A. peregrina e apoia ainda mais as aplicações medicinais desta espécie.
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Objective:To establish a method for quantitative analysis of the active ingredients including salidroside, rosarin and rosavin and content determination in Rhodiola rosea at different harvest months. Methods:HPLC was used on an X selectHSS T3 (250 mm×4.6 mm, 5 μm) column with mobile phase consisting of methol-acetonitrile-phosphoric acid (0.05%) aqueous solution for gradient elution at a flow rate of 1 ml/min. The wavelength was detected at 275 nm (salidroside) and 254 nm (rosarin, rosavin). The column temperature was set at 30 ℃ and the injection volume was 5 μl.Results:The peak areas of Salidroside, rosarin and rosavin showed good linear relationships ( r > 0.999) with the content in the ranges of 44-1 420, 10-307 and 18-573 μg, respectively. The method was precise, stable, repeatable and the sample recovery test all well satisfied the requirements of quantitative analysis. The highest accumulation of the active ingredients was observed in Rhodiola rosea in September and the content of salidroside, rosarin and rosavin were 0.66, 0.07 and 0.53 mg/g, respectively. Conclusion:This method is simple and rapid to evaluate the content of active ingredients in Rhodiola rosea.
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Objective:To establish the TLC identification method and content determination method of ferulic acid, ligustilide, hydroxysafflor yellow A and paeoniflorin in Shangke Huoxue Decoction for quality evaluation.Methods:Ferulic acid, ligustilide, hydroxysafflor yellow A and paeoniflorin were qualitatively identified by TLC method, and the content was determined by HPLC method. Waters Symmetry ShieldTM RP18 column (4.6 mm×250 mm, 5 μm) was set, the mobile phase consisted of acetonitrile-0.15% phosphoric acid water with gradient elution at a flow of 1.0 ml/min, and the column temperature was 30 ℃.The detection wavelength was 320 nm (33-50 min for ferulic acid, 55-70 min for ligustilide), 403 nm (7-31 min for hydroxysafflor yellow A) and 230 nm (7-31 min for paeoniflorin).Results:The TLC spots were clear. The linear relationships of ferulic acid, ligustilide, and hydroxysafflor yellow A were good in the range of 3.05-48.74 μg, 3.50-26.24 μg, 21.34-213.44 μg. The method was stable, repeatable with good recovery rate.Conclusion:The TLC and HPLC method for the simutanous determination of the four effective components in Shangke Huoxue Decoction were established, and the methods are suitable for the quality evaluation of Shangke Huoxue Decoction.
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A new quercetin nanocrystals self-stabilized Pickering emulsion(QT-NSSPE) was prepared by high-pressure homogenization combined with probe ultrasonic method. The influences of oil fraction, quercetin(QT) concentration, and pH of water phase on the formation of QT-NSSPE were investigated. On this basis, the QT-NSSPE prepared under optimal conditions was evaluated in terms of microstructure, stability, and in vitro release and the droplet size and drug loading were 15.82 μm and 4.87 mg·mL~(-1), respectively. The shell structure formed by quercetin nanocrystals(QT-NC) on the emulsion droplet surface was observed under a scanning electron microscope(SEM). X-ray diffraction(XRD) showed that the crystallinity of adsorbed QT-NC decreased significantly as compared with the raw QT. There were not significant changes of QT-NSSPE properties after 30 days of storage at room temperature. The in vitro release experiment confirmed that QT-NSSPE has a higher accumulative release rate than the raw QT. All these results indicated that QT-NSSPE has a great stability and a satisfactory in vitro release behavior, which is a promising new oral delivery system for QT.
Subject(s)
Emulsions/chemistry , Nanoparticles , Particle Size , Quercetin , Water/chemistryABSTRACT
Background: The information on official testing methods, or regulatory methods in Colombia to test whey in milk is limited; this restriction of information goes against the possibility of mitigating the risk of food fraud. Objectives: The validation of an HPLC method to determine casein glycomacropeptide (c-GMP), a protein that countries such as Brazil, Spain, and Ecuador have used as an indicator of raw milk adulteration with whey, was carried out. Methods: A 10mL sample of raw milk is precipitated with 24% TCA using ultrasound, a process followed by filtration. The collected fraction ensured the separation of c-GMP and then injected into the liquid chromatography. Results: A 30 minutes analysis allowed the determination of c-GMP with a retention time of 12.9 ± 0.5 minutes. The performance characteristics method in the validation exercise were: recovery percentage 99.97%, linearity R2> 0.95; % RSD accuracy <5.3%. Conclusion, the method exhibits desirable attributes for the intended purpose
Antecedentes: En Colombia la información de dominio público en metodologías de análisis de lactosuero en leche es limitada, restringiendo la posibilidad de acceder a ellas para mitigar el riesgo de fraude alimentario. Objetivos: Se realizó validación de un método por HPLC para determinar en leche cruda c-GMP, proteína usada como indicador de adulteración en países como Brasil y Ecuador. Metodos: Una muestra de 10mL de leche cruda es precipitada con TCA al 24% empleando ultrasonido, proceso seguido por filtración. La fracción recolectada aseguró la separación del c-GMP para luego inyectar al cromatógrafo líquido. Resultados: La determinación de c-GMP permitió el análisis en 30 minutos con tiempo de retención de 12,9 ± 0,5 minutos. Las características de desempeño del método en el ejercicio de validación fueron: porcentaje de recuperación 99,97%, linealidad R2>0,95; precisión %RSD< 5,3%. Conclusión: el método al final del ejercicio exhibe atributos para el fin previsto
Subject(s)
Humans , Chromatography, High Pressure Liquid , Caseins , Milk , FraudABSTRACT
With the popularization of angiography and enhanced scanning in China, high pressure contrast injector is widely used in CT, MRI, DSI and so on, the authors team try to explore and consider the harmfulness and countermeasures of the reuse of the disposable high-pressure contrast injector from the perspective of medical device supervision. Hope that it's helpful to this registration and technical evaluation.
Subject(s)
China , Contrast Media , Injections , Magnetic Resonance ImagingABSTRACT
ABSTRACT Hemoglobin A1c (HbA1c) measurement is commonly performed in diabetes mellitus patients to monitor glycemic control over the last three to four months. Carbamylated hemoglobin, which is the hemoglobin that binds to isocyanic acid derived from urea, is one of the possible analytical interference in the uremic patient. When measured by ion-exchange high-performance liquid chromatography (HPLC), carbamylated hemoglobin forms a peak that overlaps the peak of HbA1c, causing a falsely elevated HbA1c result. We report a case of a 60-years-old man who had a spurious increase in HbA1c, with a high carbamylated hemoglobin peak disproportionate to the urea value. Subsequent hemoglobin analysis using hemoglobin electrophoresis and HPLC hemoglobin testing system suggested hemoglobin J (Hb J) variant. Our case highlighted the possibility of misleading HbA1c interpretation in the presence of a high carbamylated hemoglobin peak, but not proportionate to urea value. In this study, Hb J was detected. A method free from hemoglobin variant interference should be used ideally, and monitoring glycemic control should be performed using alternative methods, such as serum fructosamine or continuous glucose monitoring.
RESUMEN La prueba de la hemoglobina glicosilada A1c (HbA1c) se realiza comúnmente en pacientes con diabetes mellitus para monitorear el control glucémico durante los últimos tres o cuatro meses. La hemoglobina carbamilada, que es la hemoglobina que se une al ácido isociánico derivado de la urea, es una de las posibles interferencias analíticas en el paciente urémico. Cuando se mide mediante cromatografía líquida de alta eficacia (HPLC) de intercambio iónico, la hemoglobina carbamilada forma un pico que se superpone al pico de HbA1c, lo que provoca un resultado de HbA1c falsamente elevado. Presentamos el caso de un hombre de 60 años que tuvo un aumento espurio de HbA1c, con un pico de hemoglobina carbamilada alto desproporcionado al valor de urea. El análisis de hemoglobina posterior mediante electroforesis de hemoglobina y el sistema de prueba de hemoglobina HPLC sugirió una variante de hemoglobina J (Hb J). Nuestro caso destacó la posibilidad de una interpretación engañosa de la HbA1c en presencia de un pico de hemoglobina carbamilada alto, pero no proporcional al valor de urea. En este estudio, se detectó Hb J. Lo ideal sería utilizar un método libre de interferencia de variantes de hemoglobina, y la monitorización del control glucémico debería realizarse mediante métodos alternativos, como la fructosamina sérica o la monitorización continua de la glucosa.
RESUMO A dosagem de hemoglobina A1c (HbA1c) é comumente realizada em pacientes com diabetes mellitus para monitorar o controle glicêmico nos últimos três a quatro meses. A hemoglobina carbamilada - hemoglobina que se liga ao ácido isociânico derivado da ureia - é uma das possíveis interferências analíticas no paciente urêmico. Quando medida por cromatografia líquida de alta eficiência (HPLC) por troca iônica, a hemoglobina carbamilada forma um pico que se sobrepõe ao pico da HbA1c, causando um resultado falsamente elevado da HbA1c. Relatamos o caso de um homem de 60 anos de idade que apresentava um aumento espúrio de HbA1c, com alto pico de hemoglobina carbamilada desproporcional ao valor de ureia. A análise subsequente da hemoglobina usando eletroforese de hemoglobina e sistema de teste de hemoglobina por HPLC sugeriu a variante de hemoglobina J (Hb J). Nosso caso destacou a possibilidade de interpretação enganosa da HbA1c na presença de um pico alto de hemoglobina carbamilada, mas não proporcional ao valor da ureia. Neste estudo, foi detectada a Hb J. Um método isento de interferência de variantes da hemoglobina deve ser idealmente usado, e o monitoramento do controle glicêmico deve ser feito usando métodos alternativos, como frutosamina sérica ou monitoramento contínuo da glicose.
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Objective:To investigate the clinical efficacy of perforator-based neurocutaneous flaps in repairing soft tissue defects of lower extremity in plateau area.Methods:Forty-eight patients who received perforator-based neurocutaneous local flaps to repair soft tissue defects of lower extremity from August, 2017 to December, 2019 were entered a follow-up study. The patients were 17 to 65 years old, 29 males and 19 females. There were 23 flaps with sural neurocutaneous perforator, 15 with saphenous neurocutaneous perforators, and 10 with superficial peroneal neurocutaneous perforators. The size of soft tissue defects ranged from 4.0 cm×3.5 cm to 15.0 cm×12.0 cm. And the size of the flaps ranged from 6.0 cm×4.5 cm to 16.0 cm×13.0 cm. The donor sites of 17 cases were directly sutured, and the rest of 31 were repaired by skin graft. The regular follow-up was performed.Results:Of the 48 cases, 33 neurocutaneous flaps survived completely, 9 had necrosis around edge of the skin after surgery and healed after hyperbaric oxygen therapy and dressing change, and 6 had complete necrosis after surgery and healed after vacuum assisted drainage and skin grafting. In 6 to 12(average, 9.6) months of follow-up after surgery, the shape and texture of flaps were in good condition with the patient satisfaction rate on functional evaluation(joint motions, standing, walking) at 75.45%.Conclusion:Perforator-based neurocutaneous flap is an excellent choice for the repair of soft tissue defects of lower extremity in plateau area because of its reliable blood supply and minimal damage to donor site.
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Introduction: Cholelithiasis is one of the most commonsurgical diseases. Laparoscopic cholecystectomy has beenuniversally accepted as gold standard for treatment ofcholelithiasis. Although many studies have shown postoperative changes in liver function tests after high pressurepneumoperitoneum during laparoscopic cholecystectomy,but very few studies have investigated the effects of lowpressure pneumoperitoneum on hepatic functions.The presentstudy was designed to compare post-operative changes inliver function tests among patients randomized to either lowpressure laparoscopic cholecystectomy(LPLC) (i.e.8 mmHg)or high pressure laparoscopic cholecystectomy(HPLC) (i.e.14mmHg).Material and methods: After taking a well informed writtenconsent,150 cases undergoing laparoscopic cholecystectomywere enrolled in the study and randomised preoperatively intotwo groups (LPLC and HPLC). Liver function tests (LFTs)including Total Bilirubin, Conjugated Bilirubin, AspartateTransaminase (AST), Alkaline Phosphatase(ALP), AlanineTransaminase (ALT), Gamma-glutamyl Transferase (GGT),Total Serum Protein, Total Serum Albumin) were done onpost-operative day (POD) 1 and day 7 and compared withpreoperative LFTs in both groups.Results: The study demonstrated significant increase inConjugated bilirubin on POD 1(p < .001), AST on POD1 (p: .005), ALT on POD 1 (p <.001), ALP on POD 1(p < .001) in HPLC as compared to LPLC patients. LFTsin both groups were found to be normal on POD 7 withoutsignificant difference between the two groups.Conclusion: Low pressure laparoscopic cholecystectomy hasless adverse effects on liver functions as compared to the highpressure laparoscopic cholecystectomy
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Objective: To prepare silymarin nanosuspension (SM-NS) with glycyrrhizic acid as stabilizer, and investigate the in vitro release characteristics and charge stabilization mechanism. Methods: SM-NS was prepared by high-speed shear-high pressure homogenization method. SM-NS lyophilized powder were prepared by freeze-drying method and characterized by physical and chemical characterization and in vitro release. The stability mechanism of SM-NS was studied from the ionic strength and pH value. Results: The dosage of glycyrrhizic acid (GA) was 0.15%. The preparation process was shear rate of 19 000 r/min, shear time of 4 min, homogenization pressure of 100 MPa, homogenization times of 12 times, and lyoprotectant was mannitol 3%, the average particle size of SM-NS lyophilized powder was (516.4 ± 10.4) nm, PDI was (0.260 ± 0.046); The in vitro release results showed that the dissolution rate and solubility of SM-NS lyophilized powder were significantly higher than the physical mixture; The study of charge stability mechanism showed that licorice acid can provide good charge stabilization and strong resistance to environmental impact. Conclusion: SM-NS is a potential and new nano-drug with high safety, which is formed by the charge stability of GA to significantly improve the solubility and stability of silymarin.
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Objective: Puerarin nanoemulsion lyophilized powder (Pue-NE-LP) was prepared using natural surfactant glycyrrhizic acid as stabilizer and evaluated in vitro. Methods: Pue-NE was prepared by high-speed shear and high-pressure homogenization method, and further combined with freeze-drying method to prepare Pue-NE-LP. Taking the average particle size and polydispersity index (PDI) as the evaluation indexes, the optimal prescription and process parameters of this experiment were screened out through a single factor test. The prepared Pue-NE-LP was characterized by physicochemical properties and dissolution in vitro. Results: The average particle size and PDI of Pue-NE-LP prepared with 5% glyceryl caprylate as oil phase, 2.0 mg/mL glycyrrhizic acid as stabilizer, and 7% glucose as lyophilization protectant was (215.1 ± 0.7) nm and (0.133 ± 0.024), respectively. Scanning electron microscopy showed that Pue-NE-LP was irregularly small and uniform in size; X-ray diffraction showed that Pue-NE-LP existed in an amorphous state. In vitro release results showed that the dissolution rate of Pue-NE-LP was significantly higher than the physical mixture. Conclusion: Pue-NE-LP prepared with natural surfactant glycyrrhizic acid as a stabilizer is not only simple to prepare, but also can significantly improve the solubility and bioavailability of puerarin. It provides a reference for the multiple development of Pue-NE formulations.