This study aims to establish a method for the determination of As B,As C,DMA,As( Ⅲ),MMA and As( Ⅴ) by using HPLC-ICP-MS. A Dioncx Ion PacTMAS7( 4 mm×250 mm) column was used for the HPLC-ICP-MS method. The mobile phase was 100 mmol·L-1 ammonium carbonate-1. 5 mmol·L-1 ammonium dibasic phosphate( gradient elution) at a flow rate of 1 m L·min-1. The injection volume was 10 μL. The linear relationships of As B,As C,DMA,As( Ⅲ),MMA,As( Ⅴ) were good with the concentration of10-500 μg·L-1. The average recovery rates( n = 6) were 105. 7%,100. 5%,102. 9%,105. 7%,100. 2%,92. 69%. The RSD were0. 50%,2. 4%,0. 93%,1. 3%,0. 89%,1. 5%. The precision and repeatability of this method were good. In this study,six forms of arsenic were separated effectively by this method. With methodological validation and sample determination,this method can be used to determine the morphological valence of arsenic in content determination.
Arsenic/analysis* ; Arsenicals/analysis* ; Chromatography, High Pressure Liquid ; Mass Spectrometry

As arsenic widely exists in nature and has been used in the pharmaceutical preparations, the traditional Chinese medicine(TCM) with arsenic include realgar(As_2S_2 or As_4S_4), orpiment(As_2S_3), and white arsenic(As_2O_3). Among the above representative medicine, the TCM compound formulas with realgar are utilized extensively. Just in Chinese Pharmacopoeia(2020 edition), there are 37 Chinese patent medicines including realgar. The traditional element analysis focuses on the detection of the total amount of elements, which neglects the study on the speciation and valence of elements. The activity, toxicity, bioavailability, and metabolic pathways of arsenic in vivo are closely related to the existence of its form, and different forms of arsenic have different effects on organisms. Therefore, the study on the speciation and valence of arsenic is of great importance for arsenic-containing TCMs and their compound formulas. This paper reviewed four aspects of the speciation and valence of arsenic, including property, absorption and metabolism, toxicity, and analytical assay.
Arsenic/analysis* ; Arsenicals/analysis* ; Sulfides ; Arsenic Trioxide ; Medicine, Chinese Traditional ; Drugs, Chinese Herbal/analysis* ; Biological Products

OBJECTIVETo develop a method for determining arsenobetaine (AsB), arsenite (AsIII), arsenate (AsV), Monomethylarsonic acid (MMA) and Dimethylarsenic acid (DMA) with liquid chromatography (LC), on-line UV-decomposition (UV), hydride generation (HG) and atomic fluorescence spectrometry (AFS) in animal origin seafood samples.

METHODSArsenic compounds were extracted in an ultrasonic bath with methanol-water (9:1) solvent from the animal origin seafood samples. The extracts were evaporated with N2 and dissolved in water. The solvent was extracted by hexane to remove lipids. And then, the aqueous solution was diluted to 10 ml. The extracts were filtered before analysis by LC-AFS. The mobile phase consisted of 0.5 mmol/L NH4H2PO4 (pH 9.0) and 20 mmol/L NH4H2PO4 (pH 6.0). Arsenic species were separated with an anion exchange column Hamilton PRPX-100 and gradient elution, detected by LC-UV-HG-AFS.

RESULTSThe established separation condition could achieve a better separation for five arsenic species. Detection Limits (LOD) were ranged from 0.0025 to 0.0032 mg/L, AsB was the predominant arsenic species in the animal origin seafood samples. AsIII and DMA were detected in certain shellfish samples at trace level. The accuracy of total arsenic measurement was tested by the analysis of NBS 1566 (Oyster Tissue). The accuracy of arsenic species measurement was tested by the analysis of BCR 627 (Tuna Fish). The data were tallied with the certified value.

CONCLUSIONArsenic species were specifically detected by LC-UV-HG-AFS in the animal origin seafood samples.

Animals ; Arsenic ; analysis ; Arsenicals ; analysis ; Chromatography, High Pressure Liquid ; methods ; Food Inspection ; methods ; Seafood ; analysis ; Spectrometry, Fluorescence ; methods

OBJECTIVETo evaluate the association between metabolism of arsenic and DNA oxidative damage in workers in a arsenic mill.

METHODSUrinary organic arsenic and 8-hydroxydeoxyguanine were detected in 37 workers highly exposed to arsenic and 16 administrative and logistic staff with mild exposure in a arsenic mill in Yunnan province, and also 28 local people who did not have the exposure in the near past time. The correlation between metabolism of arsenic and DNA oxidative damage was evaluated.

RESULTSThe urinary organic arsenic concentration was respectively (0.48 +/- 0.37) mg/L and (0.08 +/- 0.05) mg/L for men with high and low exposure, and was respectively 0.11 mg/L and (0.30 +/- 0.24) mg/ L for women with high and low exposure, while it was lower than 0.02 mg/L in the controls. Urinary 8-hydroxydeoxyguanine concentration was (18.07 +/- 11.68) micromol/mol creatinine, (11.79 +/- 8.25) micromol/mol creatinine, (10.07 +/- 3.04) micromol/mol creatinine for the males with high and low exposure and of controls, respectively, (P < 0.05), and it was 84.35 micromol/mol creatinine, (21.27 +/- 5.89) micromol/mol creatinine, (14.43 +/- 2.58) micromol/mol creatinine for females with high and low exposure and of controls, respectively. The female workers exposed to arsenic had higher urinary 8-hydroxydeoxyguanine levels than males did (P < 0.05). The increased tendencies of urinary 8-hydroxydeoxyguanine levels with the organic arsenic concentration were found in workers (r(s) = 0.279, P = 0.019).

CONCLUSIONOccupational individuals exposed to arsenic have obvious DNA oxidative damage, which is more severe in females. The difference of metabolism of arsenic may play a key role.

Adult ; Arsenicals ; urine ; China ; Female ; Guanine ; analogs & derivatives ; urine ; Humans ; Male ; Middle Aged ; Occupational Exposure ; analysis

Arsenic ; urine ; Arsenicals ; analysis ; Environmental Monitoring ; methods ; Humans ; Sensitivity and Specificity ; Spectrometry, Fluorescence ; methods

Sepiae Endoconcha is a common marine animal medicine,which generally contains high concentration of arsenic( As).The Chinese Pharmacopoeia( 2010 edition,part I) stipulated that the total As content of Sepiae Endoconcha should not exceed 2 mg·kg~(-1),while this limit was revised to 10 mg·kg~(-1) in the 2015 edition. So far,there is no research on the speciation of As in Sepiae Endoconcha,which made it hard to accurately evaluate its security risk. In this study,32 batches of Sepiae Endoconcha from different sources were collected. The safety risk assessment was carried out by determining the total As content and As speciation,inorganic As[As( Ⅲ),As( Ⅴ) ]and organic As( MMA,DMA,As C,As B) by HPLC-ICP-MS,and then the limit standard was discussed. The results showed that As B was the main form of As in Sepiae Endoconcha,followed by DMA and As( Ⅴ) ． Of the 32 batches of Sepiae Endoconcha,9 batches( accounting for 28%) were detected possessing i As. The maximum concentration of As( Ⅲ) was 103. 3 μg·kg~(-1),and the maximum concentration of As( Ⅴ) was 222. 4 μg·kg~(-1). According to the limit of i As in food,18. 75% of the samples exceeded the standard. The results indicate that there is no simple positive correlation between total As and As morphology in Sepiae Endoconcha. Besides,there is a risk in the total As limit,especially after the relaxation of the total As limit. The problem of high i As content caused by pollution and other factors is difficult to regulate. Since the toxicity of inorganic As is much higher than that of organic As,it is of great practical significance to establish inorganic As form limits in Sepiae Endoconcha.
Animals ; Arsenic/analysis* ; Arsenicals/analysis* ; Chromatography, High Pressure Liquid ; Drug Contamination ; Environmental Pollution ; Mass Spectrometry ; Medicine, Chinese Traditional ; Sepia/chemistry*

OBJECTIVE: To explore the effect of realgar on the gene expression profiles of multiple myeloma cell line RPMI 8226 by apply cDNA microarray. METHODS: The gene expression of RPMI 8226 cells before and after 48 hours of realgar treatment was determined with a cDNA microarray representing 4096 human genes. RESULTS: At the mRNA level, 164 genes were differentially altered; 53 genes were up-regulated; and 111 genes were down-regulated. CONCLUSION The realgar treatment to RPMI 8226 cell line may induce a number of gene changes. Many genes may be involved in the pathogenesis of multiple myeloma. BTG1, ALK1, and TXNIP genes may play an important role in the apoptosis and differentiation of RPMI 8226 cells.
Arsenicals ; pharmacology ; Gene Expression Profiling ; Humans ; Multiple Myeloma ; pathology ; Oligonucleotide Array Sequence Analysis ; Sulfides ; pharmacology ; Tumor Cells, Cultured

Adult ; Air Pollutants, Occupational ; adverse effects ; analysis ; Arsenicals ; adverse effects ; analysis ; Female ; Humans ; Male ; Metallurgy ; Middle Aged ; Occupational Exposure ; adverse effects ; Young Adult

BACKGROUNDHepatitis B virus (HBV) X protein (HBx) and p53 could mutually down-regulate at transcriptional level and HBx could bind with p53 protein within its transactivation domain and inhibit the function of p53 protein. In recent years, effects of arsenic trioxide (As2O3) on the expression of p53 protein have been widely studied, while little is known about the activity of p53 protein. This study was undertaken to delineate the effect of HBV X gene and As2O3 on p53 protein expression (level and activity) in HepG2 cells by small hairpin RNA (shRNA)-mediated RNA interference (RNAi) technique.

METHODSCell line HepG2 and cells with stable expression of HBV X gene (HepG2-X) were treated with 2 micromol/L As2O3, with corresponding untreated cells serving as controls. Cell lysates and nuclear extracts were extracted. Total level and the relative activity of p53 protein were detected by modified enzyme-linked immunosorbent assay (ELISA). HBV X gene sequence-specific shRNA expression vector (pXi-1 and pXi-2) and sequence-unrelated control (pXi-3) were transfected into HepG2-X. Single cell clone with stable expression of shRNA was selected and exposed to propagating culture. The effect of As2O3 on p53 protein expression and activity was re-observed.

RESULTSTotal p53 protein level was up-regulated and its relative activity ratio was enhanced by As2O3 in HepG2 and HepG2-X cells. The total p53 protein level induced by As2O3 was up-regulated by HBV X gene expression, while its relative activity was significantly suppressed. The suppression was removed after HBV X gene expression was repressed by shRNA.

CONCLUSIONSAs2O3 up-regulates p53 protein expression and enhance its activity. HBV X up-regulates As2O3 induced-p53 protein expression while suppresses its activity.

Arsenicals ; pharmacology ; Cell Line, Tumor ; Enzyme-Linked Immunosorbent Assay ; Humans ; Oxides ; pharmacology ; RNA Interference ; Trans-Activators ; genetics ; Tumor Suppressor Protein p53 ; analysis

OBJECTIVETo establish the comparative proteomic profiles of retinoid acid (RA) resistant human acute promyelocytic leukemia (APL) NB4-R1 cells before and after apoptosis induced by realgar (tetra-arsenic tetra-sulfide, As4S4).

METHODSFirst a serial of assays were performed using MTT, transmission electron microscopy, Annexin V FITC/PI double-stain, flow cytometry and confocal laser scanning microscopy to qualitatively and quantitatively observe the in vitro apoptosis inducing effect of realgar on RA-resistant cells. Then the comparative proteomic profile before and after NB4-R1 apoptosis was established using high-resolution two-dimensional electrophoresis system.

RESULTSThe inhibition effect of realgar on NB4-R1 cell growth was dose and time dependent. The 24-h 50% inhibiting concentration (IC50) was 24.06 +/- 0.19 micromol/L, and the 48-h IC50 9.50 +/- 0.13 micromol/L, and 72-h IC50 6.55 +/- 0.03 micromol/L, respectively. 24 h and 48 h were the early and late phase of major NB4-R1 apoptotic cell populations induced by 25 micromol/L realgar respectively. Differential proteomic profiles before and after realgar induced NB4-R1 apoptosis were successfully established. Averagely 1069, 975 and 893 spots could be detected of the untreated group (R0), the 24-h treatment group (R24), and the 48-h treatment group (R48), respectively by ImageMaster 2D Platinum Software. The matching rate between R24 and R0 was 79.94% and that between R48 and R0 69.33%, and that between R24 and R48 71.91%.

CONCLUSIONDifferential proteomic profiles of realgar induced NB4-R1 apoptosis were successfully established for the first time, which provided a basis for comprehensively understanding the signal transduction of realgar induced apoptosis in RA-resistant APL cells, also for screening new bio-markers and drug targets of hematopoietic malignant tumor.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; drug effects ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Leukemia, Promyelocytic, Acute ; pathology ; Proteome ; analysis ; Sulfides ; pharmacology ; Tretinoin ; pharmacology