Arsenic Speciation of Arsine-Exposed Blood Samples by High-Performance Liquid Chromatography-Inductively Coupled Plasma Mass Spectrometry and As-Adduct, A Possible Indicator of AsH3 Exposure

Abstract

Arsine (AsH(3))-exposed human blood samples were analyzed by high-performance liquid chromatography with inductively coupled plasma mass spectrometry (HPLC-ICP-MS) for arsenic speciation. After exposure of human blood samples to AsH(3) vapor for 90 min at room temperature, partial hemolysis was observed. Plasma samples from these whole blood samples were prepared by centrifugation at 1600 x g for 10 min and analyzed by HPLC-ICP-MS. In addition to arsenite [As(III); degraded from AsH(3)], an unidentified arsenic species (As-adduct) was detected at a retention time of 1.1 min. Following ultrafiltration of the plasma samples using a molecular weight cut-off of 10 kDa, As-adduct was not detected in the filtrate. To clarify the origin of As-adduct, AsH(3) was added to blank plasma and As(III) was added to both whole blood and hemolyzed blood. Although As(III) was detected in all samples, As-adduct was not detected. These results indicate that As-adduct was derived from erythrocytes during the process of hemolysis by AsH(3) and further suggest that As(III) and plasma ingredients do not contribute to As-adduct production. Therefore, the presence of As-adduct in blood could represent an indicator of acute arsine poisoning.

Speciation analysis of arsenic compounds in the serum and urine of a patient with acute arsine poisoning

“…A slight decrease was found in the sum of the concentrations of the four As species, expressed as extracted As (E-As), and the BHT was 115.5 h. When a human blood sample was exposed to arsine vapor for 90 min at room temperature, partial hemolysis was observed, and AsIII and As-adduct were detected in the plasma. However, after ultrafiltration using a cut-off of 10 kDa, the As-adduct derived from erythrocytes was not detected in the filtrate (Higashikawa et al, 2008). AsIII and erythrocyte constituents are not involved in the production of As-adduct (Higashikawa et al, 2008).…”

“…However, after ultrafiltration using a cut-off of 10 kDa, the As-adduct derived from erythrocytes was not detected in the filtrate (Higashikawa et al, 2008). AsIII and erythrocyte constituents are not involved in the production of As-adduct (Higashikawa et al, 2008). In our case, we did not detect As-adduct after ultrafiltration with a 10 kDa cut-off level, however, the ratio of E-As to T-As increased with time from 33.3% at 34 h to 53.5% at 112 h. On the contrary, constant E-As/T-As ratios were observed in the blood of a patient with acute promyelocytic leukemia (APL) after remission induction therapy using AsIII (Yoshino et al, 2009).…”

Possible production of arsenic hemoglobin adducts via exposure to arsine

“…Furthermore, a ternary DMA III -hemoglobin-haptoglobin (As-Hb-Hp) complex was detected in the plasma of rats to which arsenite was orally administered 33) . However, arsenic adduct formation was not observed in plasma when whole human blood or hemolyzed blood was mixed with trivalent inorganic arsenic in vitro 9) , and there are no reports of hemolysis due to trivalent arsenic exposure. Therefore, it is likely that trivalent arsenic does not induce hemolysis and that the arsenic adduct is not detected in plasma, although the trivalent arsenic has affinity to hemoglobin in RBCs.…”

“…Arsine was generated according to Japanese Industrial Standard (JIS) K 0102; by reducing 2 mg of arsenic trioxide with hydrochloric acid and zinc powder. The preserved blood was exposed to the generated arsine for 10 minutes as described previously 9) . Whole blood was diluted with an equal volume of Alsever's anticoagulant solution (2.05 g glucose, 0.42 g sodium chloride, 0.80 g sodium citrate, and 0.055 g citric acid per 100 ml).…”

“…Two of them Thus, a highly specific biomarker of arsine intoxication is also required. Recent reports have described an unidentified arsenic compound derived from blood cells with a molecular weight over 10,000 after in vitro exposure of blood to arsine 9) . Furthermore, it was indicated that the arsenic coexisted with hemoglobin in red blood cells separated from mice exposed to arsine 10) .…”