ABSTRACT
Background The benchmark dose (BMD) method calculates the dose associated with a specific change in response based on a specific dose-response relationship. Compared with the traditional no observed adverse effect level (NOAEL) method, the BMD method has many advantages, and the 95% lower confidence limit of benchmark dose lower limit (BMDL) is recommended to replace NOAEL in deriving biological exposure limits. No authority has yet published any health-based guideline for rare earth elements. Objective To evaluate genotoxicity threshold induced by acute exposure to neodymium nitrate in mice using BMD modeling through micronucleus test and comet assay. Methods SPF grade mice (n=90) were randomly divided into nine groups, including seven neodymium nitrate exposure groups, one control group (distilled water), and one positive control group (200 mg·kg−1 ethyl methanesulfonate), 10 mice in each group, half male and half female. The seven dose groups were fed by gavage with different concentrations of neodymium nitrate solution (male: 14, 27, 39, 55, 77, 109, and 219 mg·kg−1; female: 24, 49, 69, 97, 138, 195, and 389 mg·kg−1) twice at an interval of 21 h. Three hours after the last exposure, the animals were neutralized by cervical dislocation. The bone marrow of mice femur was taken to calculate the micronucleus rate of bone marrow cells, and the liver and stomach were taken for comet test. Results The best fitting models for the increase of polychromatophil micronucleus rate in bone marrow of female and male mice induced by neodymium nitrate were the exponential 4 model and the hill model, respectively. The BMD and the BMDL of female mice were calculated to be 31.37 mg·kg−1 and 21.90 mg·kg−1, and those of male mice were calculated to be 58.62 mg·kg−1 and 54.31 mg·kg−1, respectively. The best fitting models for DNA damage induced by neodymium nitrate in female and male mouse hepatocytes were the exponential 5 model and the exponential 4 model, respectively, and the calculated BMD and BMDL were 27.15 mg·kg−1 and 11.99 mg·kg−1 for female mice, and 16.28 mg·kg−1 and 10.47 mg·kg−1 for male mice, respectively. The hill model was the best fitting model for DNA damage of gastric adenocytes in both female and male mice, and the calculated BMD and BMDL were 36.73 mg·kg−1 and 19.92 mg·kg−1 for female mice, and 24.74 mg·kg−1 and 14.08 mg·kg−1 for male mice, respectively. Conclusion Taken the micronucleus rate of bone marrow cells, DNA damage of liver cells and gastric gland cells as the end points of genotoxicity, the BMDL of neodymium nitrate is 10.47 mg·kg−1, which can be used as the threshold of genotoxic effects induced by acute exposure to neodymium nitrate in mice.
ABSTRACT
Objective To analyze the influencing factors and benchmark value of anesthesia cost in GB25 group in CHS-DRG Group Division Strategy(1.0),and explore the cost control strategy for surgical patients.Methods The data of 152 GB25 cases in sample hospitals were analyzed statistically.Through single factor analysis and multiple stepwise regression analysis,the influencing factors of anesthesia expenses were analyzed,and the evaluation system of hospital anesthesia expenses was built according to the payment standard of GB25 group and the average level of the city.Results Univariate analysis showed that there was significant difference in the influence of laparoscopic surgery or not,operation duration and anesthesiologist on anesthesia cost(P<0.05);Stepwise regression further analysis showed that the length of operation and the anesthesiologist were the main influencing factors of anesthesia costs.Early warning shall be given when the anesthetic cost is greater than the median in the hospital,and rewards shall be given when the anesthetic cost is less than the average level in the city.Conclusion Hospitals should pay attention to standardize the diagnosis and treatment process,shorten the operation time,and reduce the anesthesia cost.At the same time,they should actively promote the clinical pathway management,and establish and improve the internal assessment mechanism of DRG.
ABSTRACT
OBJECTIVE@#The study aimed to estimate the benchmark dose (BMD) of coke oven emissions (COEs) exposure based on mitochondrial damage with the mitochondrial DNA copy number (mtDNAcn) as a biomarker.@*METHODS@#A total of 782 subjects were recruited, including 238 controls and 544 exposed workers. The mtDNAcn of peripheral leukocytes was detected through the real-time fluorescence-based quantitative polymerase chain reaction. Three BMD approaches were used to calculate the BMD of COEs exposure based on the mitochondrial damage and its 95% confidence lower limit (BMDL).@*RESULTS@#The mtDNAcn of the exposure group was lower than that of the control group (0.60 ± 0.29 vs. 1.03 ± 0.31; P < 0.001). A dose-response relationship was shown between the mtDNAcn damage and COEs. Using the Benchmark Dose Software, the occupational exposure limits (OELs) for COEs exposure in males was 0.00190 mg/m 3. The OELs for COEs exposure using the BBMD were 0.00170 mg/m 3 for the total population, 0.00158 mg/m 3 for males, and 0.00174 mg/m 3 for females. In possible risk obtained from animal studies (PROAST), the OELs of the total population, males, and females were 0.00184, 0.00178, and 0.00192 mg/m 3, respectively.@*CONCLUSION@#Based on our conservative estimate, the BMDL of mitochondrial damage caused by COEs is 0.002 mg/m 3. This value will provide a benchmark for determining possible OELs.
Subject(s)
Male , Female , Animals , Coke , Polycyclic Aromatic Hydrocarbons , DNA Copy Number Variations , Benchmarking , Occupational Exposure/analysis , DNA, Mitochondrial/genetics , DNA DamageABSTRACT
We prepared 15 batches of Kaixin Powder benchmark samples with the decoction pieces of different batches. Further, we established the specific chromatograms and index component content determination method of Kaixin Powder benchmark samples and analyzed the peaks and similarity of the chromatograms. With sibiricose A5, sibiricose A6, polygalaxanthone Ⅲ, 3,6'-disinapoyl sucrose, ginsenoside Rb_1, β-asarone, α-asarone, and dehydropachymic acid as index components, the index component content determination method was established and 70%-130% of the mean content of each component was set as the range. The chromatograms of 15 batches of Kaixin Powder benchmark samples had a total of 22 characteristic peaks, among which 8 peaks were identified, which represented sibiricose A5, sibiricose A6, polygalaxanthone Ⅲ, 3,6'-disinapoyl sucrose, ginsenoside Rb_1, β-asarone, α-asarone, and dehydropachymic acid, respectively. The chromatograms shared the similarity of 0.992-0.999. The 15 batches of benchmark samples had sibiricose A5 of 0.34-0.55 mg·g~(-1), sibiricose A6 of 0.43-0.57 mg·g~(-1), polygalaxanthone Ⅲ of 0.12-0.19 mg·g~(-1), 3,6'-disinapoyl sucrose of 1.08-1.78 mg·g~(-1), ginsenoside Rb_1 of 0.33-0.62 mg·g~(-1), β-asarone of 2.34-3.72 mg·g~(-1), α-asarone of 0.11-0.22 mg·g~(-1), and dehydropachymic acid of 0.053-0.079 mg·g~(-1). This study established the specific chromatograms and index component content determination method of Kaixin Powder benchmark samples, and the method was simple, feasible, reproducible, and stable. This study provides a scientific basis for further research on the key chemical properties of the benchmark samples and preparations of Kaixin Powder.
Subject(s)
Powders , Ginsenosides , Benchmarking , Drugs, Chinese Herbal/chemistry , Sucrose , Chromatography, High Pressure Liquid/methodsABSTRACT
Objective: To analyze correlation of occupational hydrogen fluoride exposure to low doses of bone metabolism index through occupational epidemiological investigation and benchmark dose calculation. Methods: In May 2021, using cluster sampling method, 237 workers exposed to hydrogen fluoride in a company were selected as the contact group, and 83 workers not exposed to hydrogen fluoride in an electronics production company were selected as the control group. The external exposure dose and urinary fluoride concentration, blood and urine biochemical indicators of the workers was measured.The relationship between external dose and internal dose of hydrogen fluoride was analyzed. The external dose, urinary fluoride was used as exposure biomarkers, while serum osteocalcin (BGP), serum alkaline phosphatase (AKP) and urinary hydroxyproline (HYP) were used as effect biomarkers for bone metabolism of hydrogen fluoride exposure. The benchmark dose calculation software (BMDS1.3.2) was used to calculate benchmark dose (BMD) . Results: Urine fluoride concentration in the contact group was correlated with creatinine-adjusted urine fluoride concentration (r=0.69, P=0.001). There was no significant correlation between the external dose of hydrogen fluoride and urine fluoride in the contact group (r=0.03, P=0.132). The concentrations of urine fluoride in the contact group and the control group were (0.81±0.61) and (0.45±0.14) mg/L, respectively, and the difference between the two groups was statistically significant (t=5.01, P=0.025). Using BGP, AKP and HYP as effect indexes, the urinary BMDL-05 values were 1.28, 1.47 and 1.08 mg/L, respectively. Conclusion: Urinary fluoride can sensitively reflect the changes in the effect indexes of biochemical indexes of bone metabolism. BGP and HYP can be used as early sensitive effect indexes of occupational hydrogen fluoride exposure.
Subject(s)
Humans , Fluorides/adverse effects , Hydrofluoric Acid , Benchmarking , Biomarkers , Occupational Exposure/adverse effectsABSTRACT
Objective To explore the biological exposure limit of blood system damage caused by long-term exposure to polycyclic aromatic hydrocarbons (PAHs) in non-occupational population by using the benchmark dose method, and to provide relevant reference for further improving the assessment of PAHs-induced health damage effects. Methods Adult residents living in downwind direction of a coke-oven plant in Shanxi Province were selected as the research subjects, and the information collected from baseline was used as the control. The metabolites of PAHs in urine were used as exposure biomarker, and the abnormal rate of red blood cell index was used as response biomarker. The relationship between urinary OH-PAHs and the erythrocyte damage rate was analyzed, and the benchmark dose (BMD) and the lower confidence limitation for the benchmark dose (BMDL) were calculated using Bayesian dose-optimizing software. Results The urinary PAH metabolites were mainly naphthalene and fluorene. The detection concentrations of 2-OHFlu and 1-OHPhe in the final year were higher than those in the baseline (P<0.05). With the increase of exposure years, the abnormal rate of red blood cells in the final year was higher than that in the baseline (P<0.05). In addition, the abnormal rate of red blood cells increased with the increase of the concentrations of five metabolites of PAHs in urine, and the chi-square trend test was significant (P<0.05). The benchmark dose (BMD) of OH-PAHs was 0.67 μmol/mol Cr, 0.82 μmol/mol Cr, 1.40 μmol/mol Cr and 0.78 μmol/mol Cr, respectively. The BMD of 2-OHNap in people with barbecue diet habits was 0.23 μmol/mol Cr. The BMD of 2-OHNap in people without barbecue diet habits was 1.44 μmol/mol Cr. Conclusion There is a dose-response relationship between the concentration of PAHs metabolites in urine and the damage of red blood cells. Long-term exposure to PAHs can lead to hematological damage. It is suggested that targeted public health interventions should be formulated to reduce the exposure of the general population to PAHs.
ABSTRACT
ObjectiveTo establish a high performance liquid chromatography(HPLC) fingerprint of Yanghetang benchmark sample, and evaluate its quality with chemometric methods, so as to provide a reference for the quality control of this benchmark sample. MethodHPLC was used to establish the fingerprint of Yanghetang benchmark sample with ZORBAX SB-C18 column(4.6 mm×250 mm, 5 μm), the mobile phase was consisted of acetonitrile(A) -0.05% phosphoric acid aqueous solution (containing 0.05% triethylamine solution)(B) for gradient elution(0-5 min, 2%-3%A; 5-15 min, 3%-5%A; 15-65 min, 5%-30%A; 65-90 min, 30%-70%A), the flow rate was 1.0 mL·min-1, the column temperature was 35 ℃, and the detection wavelength was 210, 260 nm. Traditional Chinese Medicine(TCM) Chromatographic Fingerprint Similarity Evaluation System (2012 edition) combined with cluster analysis, principal component analysis(PCA) and partial least squares-discriminant analysis(PLS-DA) were used to evaluate the quality differences between different batches of Yanghetang benchmark samples, and to find the main chemical components responsible for the quality differences. ResultHPLC fingerprint of Yanghetang benchmark sample was established, 13 common peaks were identified and attributed to each common peak, including peaks 2 and 8 from Rehmanniae Radix Praeparata, peaks 10 and 11 from Cinnamomi Cortex, peaks 1, 3-6 from fried Sinapis Semen, peak 13 from Ephedrae Herba, and peaks 7, 9, 12 from Glycyrrhizae Radix et Rhizoma. Eight of them were identified by comparing with control substance, which were 5-hydroxymethylfurfural(peak 2), sinapine thiocyanate(peak 4), glycyrrhizin(peak 7), verbascoside(peak 8), cinnamic acid(peak 10), cinnamaldehyde(peak 11), glycyrrhizic acid(peak 12) and ephedrine hydrochloride(peak 13). The similarities of the HPLC fingerprints of 15 batches of Yanghetang benchmark samples with the control fingerprint were all greater than 0.80. The three chemometric methods could classify the samples into two categories. Eight differential components were screened out, among which 5-hydroxymethylfurfural, sinapine thiocyanate, verbascoside and ephedrine hydrochloride were identified. ConclusionThe established fingerprint analysis method is accurate, stable and reproducible, which basically reflects the overall chemical composition of Yanghetang benchmark sample, and can provide a basis for establishment of quality standards for compound preparations of this famous classical formula.
ABSTRACT
ObjectiveTo establish the specific chromatogram and thin layer chromatography(TLC) of Qingxin Lianziyin(QXLZY) benchmark samples, in order to clarify the key quality attributes and provide a reference for the quality evaluation of QXLZY. MethodHigh performance liquid chromatography(HPLC) specific chromatogram of QXLZY benchmark samples was developed by using a YMC Hydrosphere C18 column(4.6 mm×250 mm, 5 μm) with the mobile phase of acetonitrile(A)-0.2% formic acid aqueous solution(B) for gradient elution(0-10 min, 5%-20%A; 10-20 min, 20%A; 20-25 min, 20%-24%A; 25-40 min, 24%-30%A; 40-55 min, 30%-50%A; 55-65 min, 50%-100%A; 65-75 min, 100%A; 75-75.1 min, 100%-5%A; 75.1-90 min, 5%A), and the detection wavelength was 360 nm. Ultra-high performance liquid chromatography-linear ion trap/orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS) with electrospray ionization(ESI) was used to identify the components of QXLZY benchmark samples by accurate relative molecular weight and multilevel MS fragment ion information, the detection conditions were positive and negative ion modes and data dependency scanning mode. TLC identification methods for Ophiopogonis Radix, Lycii Cortex, Nelumbinis Semen, Poria, Astragali Radix and Ginseng Radix et Rhizoma in QXLZY were established. ResultA total of 15 characteristic peaks were identified from Glycyrrhizae Radix et Rhizoma, Plantaginis Semen and Scutellariae Radix, and the relative standard deviations of the retention times of 15 characteristic peaks in 15 batches of QXLZY benchmark samples were≤3% with peak 8(baicalin) as the reference peak. A total of 100 compounds, including flavonoids, organic acids, saponins, amino acids and others, were identified in the benchmark samples by UHPLC-LTQ-Orbitrap MS. The established TLC had good separation and was suitable for the identification of Ophiopogonis Radix, Lycii Cortex, Nelumbinis Semen, Poria, Astragali Radix and Ginseng Radix et Rhizoma in QXLZY. ConclusionThe material basis of QXLZY benchmark samples is basically determined by MS designation and source attribution. The established specific chromatogram and TLC of QXLZY are simple, stable and reproducible, which can provide a reference for the development and quality control of QXLZY.
ABSTRACT
ObjectiveBased on the supramolecular "imprinting template" theory, the autonomous action law of the component groups of Shentong Zhuyutang in the preparation process of medicinal materials-decoction pieces-formulas was studied to clarify the quantitative transfer law of its quality attributes. MethodUltra performance liquid chromatography(UPLC) fingerprint of Shentong Zhuyutang was established with mobile phase of 0.4% phosphoric acid aqueous solution(A)-acetonitrile(B) for gradient elution(0-2.5 min, 100%A; 2.5-6 min, 100%-96%A; 6-15 min, 96%-92%A; 15-25 min, 92%-88%A; 25-35 min, 88%-75%A; 35-50 min, 75%-65%A; 50-60 min, 65%-50%A; 60-65 min, 50%-30%A; 65-70 min, 100%A) and detection wavelength of 235 nm, and the total statistical moments, information entropy and primary feeding amount of fingerprint of medicinal materials, decoction pieces and benchmark samples were calculated. Dry extract rate of the benchmark samples, the transfer rates and the addition parameters of medicinal materials-decoction pieces-formulas were calculated. ResultSimilarities of the total statistical moments of UPLC fingerprint of 15 batches of medicinal materials and decoction pieces were>0.89, the relative standard deviations(RSDs) of information entropy of UPLC fingerprint of 12 medicinal materials and decoction pieces were<10%. RSDs of total first-order moment(MCRTT) and information entropy of Shentong Zhuyutang(medicinal materials) were 5.5% and 2.3%, while the RSDs of MCRTT and information entropy of Shentong Zhuyutang(decoction pieces) were 4.8% and 2.6%, respectively. The dry extract rate of 45 batches of Shentong Zhuyutang was 17.2%-20.2%. The transfer rate of medicinal materials to decoction pieces was within the range of data fluctuation, which was 70%-130% of the average value. The overall transfer rates of medicinal materials to decoction pieces and decoction pieces to benchmark samples were 101.8% and 83.0%, respectively. ConclusionThe quality properties of Shentong Zhuyutang benchmark samples can be studied by total statistical moment analysis and primary feeding amount analysis, which can confirm the supramolecular "imprinting template" theory to a certain extent.
ABSTRACT
Proteins equipped with flavin adenine dinucleotides (FAD) or flavin mononucleotides (FMN) are named flavoproteins and constitute about 1% of all existing proteins. They catalyze redox, acid-base and photochemical reactions in a variety of biochemical phenomena that goes from energy metabolism to DNA repair and light sensing. The versatility observed in flavoproteins is ultimately a balance of flavin intrinsic properties modulated by a protein environment. This thesis aims to investigate how flavoproteins work by systematic evaluating flavin properties and reactivity. In particular, the mechanism of fumarate reduction by the flavoenzyme fumarate reductase Fcc3 was determined. Electronic-structure calculations were used for this task based on rigorous calibration with experimental data and error assessment. Flavin properties at chemical accuracy were obtained with single reference coupled-cluster CCSD(T) calculations at the complete basis set limit. Density functional theory was demonstrated an excellent alternative with lower computational costs and slightly less accuracy. Flavin protonation and tautomerism were shown to be important modulators of flavin properties and reactivity, with the possibility of various tautomers existing at neutral pH. Regarding flavin redox properties, an analysis based on multiconfigurational wave function weights was proposed for categorizing flavin redox reactions as hydride or hydrogen-atom transfers. This analysis is an upgrade over traditional partial charges methods and can be applied not only to flavin reactions but to any protoncoupled electron transfer. In the investigation of the enzymatic mechanism of fumarate reduction, the reaction was determined as a nucleophilic addition by hydride transfer with carbanion formation. Fumarate reductase employs electrostatic catalysis in contrast to previous proposals of substrate straining and general-acid catalysis. Also, hydride transfer was shown to be vibronically adiabatic with low tunneling contribution. These findings give new insights into the mechanisms of fumarate reductases and provide a framework for future computational studies of flavoproteins in general. The analyses and benchmark studies presented can be used to build better models of properties and reactivity of flavins and flavoproteins
Proteínas equipadas com dinucleotídeos de flavina-adenina (FAD) e mononucleotídeos de flavina (FMN) são chamadas flavoproteínas e constituem cerca de 1% de todas as proteínas existentes. Elas catalisam reações redox, ácido-base e fotoquímicas numa variedade de fenômenos bioquímicos que vão desde o metabolismo energético até reparo de DNA e captação de luz. A versatilidade observada em flavoproteínas é em última instância um balanço das propriedades intrínsecas de flavinas moduladas por um ambiente proteico. Esta tese busca investigar como flavoproteínas funcionam através de avaliações sistemáticas de propriedades e reatividade de flavinas. Em particular, o mecanismo de redução de fumarato pela flavoenzima fumarato redutase Fcc3 foi determinado. Cálculos de estrutura eletrônica foram usados para esta tarefa com base em rigorosa calibração com dados experimentais e avaliação de erros. As propriedades de flavinas foram determinadas com acurácia química com cálculos monoconfiguracionais de coupled-cluster CCSD(T) no limite de conjunto base completo. A teoria do funcional da densidade mostrou-se uma alternativa excelente com menor custo computacional e um pouco menos de acurácia. Protonação e tautomerismo de flavinas mostraram-se moduladores importantes de suas propriedades e reatividade, com a possibilidade de vários tautômeros existirem em pH neutro. Em relação às propriedades redox de flavinas, uma análise baseada nos pesos de funções de onda multiconfiguracionais foi proposta para categorizar as reações redox de flavinas como transferências de hidreto ou hidrogênio. Esta análise é uma melhoria em relação aos métodos tradicionais de cargas parciais e pode ser aplicada não apenas para reações de flavinas mas para qualquer transferência de próton acoplada a elétrons. Na investigação do mecanismo enzimático de redução de fumarato, a reação foi designada como uma adição nucleofílica por transferência de hidreto e formação de carbânion. A fumarato redutase usa catálise eletrostática diferentemente de prospostas anteriores envolvendo distorção do substrato e catálise ácida geral. Além disso, a transferência de hidreto mostrou-se vibronicamente adiabática com pouca contribuição de tunelamento. Estas descobertas abrem novas perspectivas sobre os mecanismos de fumarato redutases e fornecem uma base para estudos computacionais futuros sobre flavoproteínas em geral. As análises e estudos comparativos apresentados podem ser usados para construir melhores modelos para propriedades e reatividade de flavinas e flavoproteínas
Subject(s)
Comparative Study , Flavins/analysis , Flavoproteins/analysis , Calculi/chemistry , Static Electricity/adverse effects , FumaratesABSTRACT
Objective@#The present study was undertaken to evaluate the subchronic oral toxicity of sodium dehydroacetate (DHA-Na) and to determine the point of departure (POD), which is a critical factor in the establishment of an acceptable dietary intake.@*Methods@#DHA-Na was administered once daily by gavage to Sprague-Dawley rats at dose levels of 0.0, 31.0, 62.0, and 124.0 mg/kg BW per day for 90 days, followed by a recovery period of 4 weeks in the control and 124.0 mg/kg BW per day groups. The outcome parameters were mortality, clinical observations, body weights, food consumption, hematology and clinical biochemistry, endocrine hormone levels, and ophthalmic, urinary, and histopathologic indicators. The benchmark dose (BMD) approach was applied to estimate the POD.@*Results@#Significant decreases were found in the 62.0 and 124.0 mg/kg BW groups in terms of the body weight and food utilization rate, whereas a significant increase was found in the thyroid stimulating hormone levels of the 124.0 mg/kg BW group. Importantly, the 95% lower confidence limit on the BMD of 51.7 mg/kg BW was modeled for a reduction in body weight.@*Conclusion@#The repeated-dose study indicated the slight systemic toxicity of DHA-Na at certain levels (62.0 and 124.0 mg/kg BW) after a 90-day oral exposure.
Subject(s)
Animals , Rats , Body Weight , Organ Size , Pyrones , Rats, Sprague-DawleyABSTRACT
Fingerprints of 18 batches of substance benchmark of Shentong Zhuyu Decoction(SZD) were established by UPLC under the following conditions: Waters Sun Fire C_(18) column(3.0 mm×150 mm, 3.5 μm), column temperature of 35 ℃, gradient elution with mobile phase of acetonitrile(A)-0.1% phosphoric acid aqueous solution(B) at the flow rate of 0.4 mL·min~(-1), and detection by wavelength switching. A total of 16 common peaks were identified. The similarities among the fingerprints were calculated by Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012 Edition) and the result showed they were in the range of 0.911-0.988. Based on the 16 common peaks, cluster analysis(CA), principal component analysis(PCA), and partial least square discriminant analysis(PLS-DA) all categorized the 18 batches of samples into two groups(S1, S2, S5-S8, S14, and S17 in one group, and S1, S2, S5-S8, S14, and S17 in another), and 11 most influential components were screened. Five known components with great difference among samples(hydroxysafflor yellow A, ferulic acid, benzoic acid, ecdysone, and ammonium glycyrrhizinate) were determined. The combination of multi-component content determination and fingerprints can reflect the overall cha-racteristics of the primary standards of SZD, which is simple, feasible, reproducible, and stable. This study can serve as a reference for the quality control of the primary standards of SZD.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/standards , Quality ControlABSTRACT
Ultra-performance liquid chromatography-quadrupole-electrostatic field Orbitrap mass spectrometry(UHPLC-Q-Exactive Orbitrap MS/MS) was used for rapid identification of the chemical components in Kaixin San substance benchmark. The gradient elution was performed through a Waters ACQUITY~(TM) BEH C_(18) column(2.1 mm×150 mm, 1.7 μm) with water-acetonitrile as mobile phase, a column temperature of 30 ℃, a flow rate of 0.3 mL·min~(-1), and a sample size of 1 μL. The scanning was performed in the negative ion mode. The complex component groups in Kaixin San substance benchmark were quickly and accurately identified and clearly assigned based on the comparison of the retention time and MS data with those of the reference substance as well as the relative molecular weight of the same or similar components in the mass spectrum database and literature. A total of 77 compounds were identified, including 26 saponins, 13 triterpenoid acids, 20 oligosaccharide esters, 5 xanthones, and 13 other compounds. The qualitative method established in this study can systematically, accurately, and quickly identify the chemical components in Kaixin San substance benchmark, which can provide a basis for the further analysis of its active components in vivo and the establishment of its quality control system.
Subject(s)
Benchmarking , Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/chemistry , Tandem Mass Spectrometry/methodsABSTRACT
The methods for determining the characteristic chromatogram and index components content of Xuanfu Daizhe Decoction were established to provide a scientific basis for the quality evaluation of substance benchmarks and preparations. Eighteen batches of Xuanfu Daizhe Decoction were prepared with the decoction pieces of different batches and of the same batch were prepared respectively, and the HPLC characteristic chromatograms of these samples were established. The similarities of the chromatographic fingerprints were analyzed. With liquiritin, glycyrrhizic acid, 6-gingerol, ginsenoside Rg_1, and ginsenoside Re as index components, the high performance liquid chromatography was established for content determination with no more than 70%-130% of the mass average as the limit. The results showed that there were 19 characteristic peaks corresponding to the characteristic chromatograms of 18 batches of Xuanfu Daizhe Decoction, including 8 peaks representing liquiritin, 1,5-O-dicaffeoylqunic acid, ginsenoside Rg_1, ginsenoside Re, 1-O-acetyl britannilactone, ginsenoside Rb_1, glycyrrhizic acid, and 6-gingerol, and the fingerprint similarity was greater than 0.97. The contents of liquiritin, glycyrrhizic acid, 6-gingerol, and ginsenosides Rg_1 + Re in the prepared Xuanfu Daizhe Decoction samples were 0.53%-0.86%, 0.61%-1.2%, 0.023%-0.068%, and 0.33%-0.66%, respectively. Except for several batches, most batches of Xuanfu Daizhe Decoction showed stable contents of index components, with no discrete values. The characteristic chromatograms and index components content characterized the information of Inulae Flos, Ginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma, and Zingiberis Rhizoma Recens in Xuanfu Daizhe Decoction. This study provides a scientific basis for the further research on the key chemical properties of substance benchmark and preparations of Xuanfu Daizhe Decoction.
Subject(s)
Benchmarking , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/chemistry , Ginsenosides/analysis , Glycyrrhizic Acid/analysis , Quality ControlABSTRACT
Objective To investigate the situation of drinking water fluorosis in Tianjin, to study the benchmark dose of urinary fluoride, and to provide a reference for the formulation of the standard for urinary fluoride. Methods Three fluorosis endemic areas were selected in Tianjin, and three villages in each area were selected in this study. The water fluoride was detected, and the prevalence of dental fluorosis and the urinary fluoride of children aged 8-12 years old were investigated. Results The prevalence of dental fluorosis and urinary fluoride content in children in the villages with water fluoride exceeding the standard were significantly higher than those in the villages with water fluoride qualified, and the difference was statistically significant (χ2=88.821, P<0.001; Z=6.089, P<0.001). The analysis of the prevalence of dental fluorosis in children showed that the younger the age, the lower the prevalence and severity of dental fluorosis (χ2trend=14.584, P2trend= 20.525, P2=0.736, P=0.391; χ2=3.649, P=0.456). There were significant differences in urinary fluoride between children with different ages and genders (H=14.768, P=0.011; Z=-2.526, P=0.012). According to the level of urinary fluoride, the children were divided into 5 groups. With the increase of urinary fluoride concentration, the prevalence of dental fluorosis gradually increased and the difference was statistically significant (χ2trend=16.348, P<0.001), showing a dose-effect relationship. Based on the prevalence of dental fluorosis in children, BMDS was used to calculate the BMD and BMDL, which were 2.20 mg/L and 1.54 mg/L, respectively. Conclusion The water reform and fluoride reduction in Tianjin has achieved certain results. There is a dose-effect relationship between urinary fluoride and prevalence of dental fluorosis in children, and the reference dose value is slightly higher than the current standard, suggesting that the current standard can be updated timely.
ABSTRACT
ObjectiveTo establish the determination for index components in benchmark samples of Erdongtang, and clarify the content and transfer rate rages of index components in 15 batches of benchmark samples, and to explore the quantity transfer of index components of decoction pieces to benchmark samples. MethodFifteen batches of benchmark samples were prepared, the contents of mangiferin, baicalin and glycyrrhizic acid were determined by high performance liquid chromatography (HPLC)-diode array detector (DAD), the mobile phase was acetonitrile (A)-0.1% formic acid aqueous solution (B) for gradient elution (0-10 min, 10%-17%A; 10-25 min, 17%-19%A; 25-28 min, 19%-25%A; 28-45 min, 25%-33%A; 45-46 min, 33%-45%A; 46-60 min, 45%-55%A), detection wavelength was set at 254 nm. Contents of timosaponin BⅡ and the sum of protoneodioscin and protodioscin were determined by HPLC-evaporative light scattering detector (ELSD), the mobile phase was acetonitrile (A)-water (B) for gradient elution (0-20 min, 24%A; 20-25 min, 24%-27%A; 25-33 min, 27%-28%A; 33-36 min, 28%-90%A; 36-41 min, 90%-24%A). ResultThe methodological verification of the established method was good, which could be used for determination of five index components in benchmark samples. The content ranges of mangiferin, baicalin, glycyrrhizic acid, timosaponin BⅡ, and the sum of protoneodioscin and protodioscin in 15 batches of benchmark samples of Erdongtang were 0.14%-0.23%, 2.40%-3.37%, 0.07%-0.44%, 0.43%-0.95%, and 0.15%-0.47%, the transfer rate ranges of them were 33.90%-52.15%, 84.46%-105.61%, 22.59%-93.86%, 38.07%-61.43%, and 53.28%-96.11%, respectively. ConclusionThe consistencies of transfer rate of mangiferin, baicalin, timosaponin BⅡ and the sum of protoneodioscin and protodioscin (except glycyrrhizic acid) between decoction pieces and benchmark samples of Erdongtang are good, indicates that the transfer rates of 4 index components are stable during the preparation process of benchmark samples, which can provide data support for research and development of the compound preparation of this formula.
ABSTRACT
ObjectiveTo establish the quality standard of Liangditang benchmark samples. MethodUltra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) was used to qualitatively analyze the chemical composition of Liangditang on the basis of molecular and fragment ion peak information with cracking law. The mobile phase was methanol (A)-0.05% phosphate aqueous solution (B) for gradient elution (0-10 min, 5%-23.5%A; 10-20 min, 23.5%A; 20-58 min, 23.5%-63%A; 58-60 min, 63%-90%A), the flow rate was 0.8 mL·min-1, and the detection wavelength was 254 nm. Electrospray ionization was employed under positive ion mode, the detection range was m/z 100-1 700. Key quality attributes and sources were determined by comparing with single medicine and reference substances. Through mass transfer analysis of multiple batches from decoction pieces to benchmark samples, high performance liquid chromatography (HPLC) for determining the contents of index components and HPLC detection of characteristic maps were established. Through the determination of 15 batches of benchmark samples, the content range of the index components and the common peaks of the characteristic map were determined. Thin layer chromatography (TLC) was applied to the identification of 5 medicines in the formula. Moisture and dry extract yield of the benchmark samples were determined by drying method. ResultA total of 27 compounds were inferred from the benchmark samples of Liangditang, among which 9 compounds were confirmed by comparison with the control, including catalpol, harpagide, gallic acid, albiflorin, paeoniflorin, verbascoside, angoroside C, cinnamic acid and harpagoside. A method for determining the characteristic maps of the benchmark samples were established and 13 peaks were assigned, and the characteristic peaks were mainly derived from wine-processed products of Rehmanniae Radix, Scrophulariae Radix and wine-processed products of Paeoniae Radix Alba. The similarity between the characteristic map of 15 batches of benchmark samples and the control characteristic map was >0.9. Methods for the determination of paeoniflorin, harpagoside, L-hydroxyproline and glycine were established, and the contents of these four components in 15 batches of benchmark samples were within ±30% of the corresponding mean value, and the transfer rate of decoction pieces to the benchmark samples was stable and controllable. TLC was established to identify 5 prescription drugs (except Ejiao) with two kinds of test solutions, and the results showed that the method had good specificity. The average dry extract yield was 48.06%, and the average moisture was 5.58%, which were within the range of ±10% and ±30% of their mean values, respectively. ConclusionThe quality standard of Liangditang benchmark samples was as follows:the similarity between the benchmark samples and the control characteristic map is >0.9, the contents of paeoniflorin, harpagoside, L-hydroxyproline and glycine are 217-403, 24-46, 634-1 178, 1 253-2 328 mg per dose, the dry extract yield is 43.0%-53.0%, the moisture is 4.0%-7.0%, under the set detection conditions, the benchmark samples have corresponding characteristic spots by comparing with the control herbs of 5 medicines. This quality standard is stable and reliable, which fills the gap in the quality control of Liangditang, and can provide a reference for the establishment of the quality standard of Liangditang granules.
ABSTRACT
Background The optimal model method for estimation of benchmark dose (BMD) does not consider the uncertainty of model selection. There is a lack of studies on using Bayesian model averaging (BMA) to estimate BMD. Objective To apply BMA to the exposure assessment of cadmium pollution in China, discuss the role of BMA in estimating BMD based on dose-response models, and to provide methodological support for health risk assessment of hazardous substances. Methods The parameters of five dose-response models (Gamma, Log-logistic, Log-probit, Two-stage, and Weibull models) estimated from the data from a cadmium-contaminated area in Baiyin City of Gansu Province and the urinary cadmium ranges in five cadmium-contaminated areas in China were used to simulate the data of varied correct models with different numbers of dosage groups (5 and 8) and different sample sizes (50, 100, and 200), then the performance of BMA and traditional optimal model were compared. The case analysis used the cadmium exposure data in Baiyin, Gansu Province. All analyses set urinary cadmium as the indicator of cadmium exposure, the abnormal rate of β2-microglobulin as the effect indicator, and the benchmark response to 10%. The correct model (the model used when simulating data), optimal model [the model with smallest Akaike information criterion (AIC)], and BMA were used to estimate BMD and lower confidence limit of benchmark dose (BMDL); the BMDs, BMDLs, and relative deviations from different methods were compared. Results In the simulation study, with increasing sample size or the number of dosage groups, the intervals of the 5th percentile and the 90th percentile of BMD tended to be narrower; when the correct model was a single model, the relative deviation of BMD estimation by BMA was greater than that of the traditional optimal model; when the correct model was an equal weight mixed model, the relative deviation of BMD estimation by BMA was less than that by the traditional optimal model. For the data of cadmium-contaminated areas, the optimal model was a Log-probit model (AIC=1814.46), followed by a Log-logistic model (AIC=1814.57); the BMDs (BMDLs) estimated by the Log-probit model, the Log-logistic model, and BMA were 3.46 (2.68), 3.16 (2.33), and 2.92 (2.07) μg·g−1, respectively. Conclusion The traditional optimal model is still recommended when the correct model is known. However, when the dose-response relationship of a hazardous substance is uncertain or with different sources or exposure grouping, compared with the traditional optimal model, BMA theoretically provides more stable estimation of BMD and BMDL by considering multiple possible alternative models.
ABSTRACT
By preparing 15 batches of lyophilized powder samples of substance benchmark in Houpo Wenzhong Decoction,the fingerprint,index component content and extract rate were determined,and the characteristic peaks,the range of similarity with the reference map,the content range and transfer rate range of magnolol,hesperidin,glycyrrhizic acid and pinocembrin,the extract rate range and the change range were clarified. The results showed that the similarity between the fingerprint of substance benchmark and the reference map R generated from the 15 batches of substance benchmark samples was higher than 0. 90. The assignment of the characteristic peaks in the full prescription's fingerprint of the herbs except Poria cocos was clarified. Nineteen characteristic peaks were assigned,and 12 characteristic peaks were assigned by the reference substance,of which 4 were from Magnolia ocinalis Cortex,5 from Exocarpium Citri Rubrum,2 from Radix aucklandiae,3 from Glycyrrhiza Radix et Rhizoma,4 from Semen Alpiniae Katsumadai,and one from Rhizoma Zingiberis and Zingiber officinale Roscoe. The index component content range and transfer rate range were 0. 80%-1. 14% and 20. 25%-39. 61% for hesperidin,0. 49%-0. 79% and 23. 09%-33. 87%for glycyrrhizic acid,0. 03%-0. 07% and 3. 55%-10. 09% for pinocembrin,0. 15%-0. 38% and 8. 08%-24. 35% for magnolol. The extract rate range and the change range were22. 60%-25. 57% and 12. 67%-23. 68% respectively. In this study,we introduced the concepts of index component content,fingerprint,extract rate,explored the transfer relation of quality value transmitting of substance benchmark in Houpo Wenzhong Decoction,and initially established the quality standard of Houpo Wenzhong Decoction,all of which would provide ideas for the development and research of similar prescriptions.
Subject(s)
Benchmarking , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Glycyrrhiza , Quality ControlABSTRACT
Based on the textual research on literature, the key information of Wenjing Decoction were tested and identified, and 15 batches of lyophilized powder samples of Wenjing Decoction were prepared. The specific components, including paeoniflorin, glycyrrhizin, ginsenosides(Rg_1, Re and Rb_1), glycyrrhizic acid, and paeonol, were used as indexes to establish the HPLC method for quantitative evaluation, and the content ranges and transfer rates of these components were determined. The results showed that the contents of paeoniflorin, glycyrrhizin, ginsenosides Rg_1 + Re, ginsenoside Rb_1, glycyrrhizic acid, and paeonol in the 15 batches of samples were 0.62%-0.86%, 0.25%-0.76%, 0.14%-0.30%, 0.07%-0.21%, 0.63%-1.16%, and 0.09%-0.25%, respectively, and their transfer rates from the decoction pieces to the reference materials were 14.99%-19.42%, 28.11%-40.93%, 25.92%-61.88%, 25.03%-64.06%, 23.43%-35.53%, and 5.34%-10.44%, respectively. The consistency of the transfer rates between batches indicated that the preparation process was stable. It is suggested that the contents of paeoniflorin, glycyrrhizin, ginsenosides Rg_1 + Re, ginsenoside Rb_1, glycyrrhizic acid, and paeonol in Wenjing Decoction should not be less than 0.52%, 0.35%, 0.15%, 0.10%, 0.63%, and 0.12%, respectively. In this study, we determined the contents and analyzed the quantity transfer process of the index components in Wenjing Decoction, which can provide a basis for the follow-up development of Wenjing Decoction and the quality control of related preparations.