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An autopsy case of acute selenium (selenious acid) poisoning and selenium levels in human tissues
87
Forensic Science International, 31 (1986) 87-9 2 Elsevier Scientific Publishers Ireland Ltd.
AN AUTOPSY CASE OF POISONING AND SELENIUM
ACUTE LEVELS
SELENIUM (SELENIOUS IN HUMAN TISSUES
RYOJI MATOBA, HIROSHI KIMURA, EIKOH UCHIMA*, YAMADAa, YOICHI MITSUKUNIa and ICHIRO SHIKATA
TAKASHI
Department of Legal Medicine, Faculty of Medicine, Osaka Medical Examiner’s Office, Nakanoshima, qsaka 530 (Japan)
ACID)
ABE, TADASHI
University
and
“Osaka
(Received November 4, 1985) (Revision received December 23, 1985) (Accepted January 17, 1986)
Summary A case of fatal suicidal ingestion of “Super Blue (Gun Blue)” (gun-blueing) is presented. Post-mortem examination of the patient revealed pulmonary edema with pleural effusion and congestion of the kidney. Necrosis of proximal tubules was found in the kidney by histological examination. “Super Blue” contains 4% selenious acid and 2.5% cupric sulfate in HCl. Levels of selenium and copper in tissues of the toxic case and normal individuals were determined. The selenium levels of tissues of the patient were 9 -90-fold higher than that of normal subjects, whereas concentrations of copper were about 2-fold compared to that of control levels. The highest levels of selenium in the tissues of the patient were found in the lung, kidney and stomach contents. Key
words:
Selenium poisoning; Tissue selenium; Autopsy
Introduction Selenium is one of the essential trace elements in human and animal nutrition [l-3] and is known as an integral part of the enzyme of glutathione peroxidase of human erythrocytes [4]. It is known that 0.04 -0.1 ppm of selenium is required in the diet for health, while a deficient or toxic effect appears if the diet contains low selenium or more than 4 ppm, respectively [l]. Selenium deficiency is known as “white muscle disease” for animals and Keshan disease, an endemic cardiomyopathy in China, for humans [2]. Some relationships between selenium deficiency and human cardiovascular diseases have been suggested from epidemiological studies [5]. Acute and *Present address: Department of Legal Medicine, School of Medicine, University of the Ryukyus, Nishihara, Okinawa 90301, Japan. Address all correspondence to: Dr. Ryoji Matoba, Department of Legal Medicine, Faculty of Medicine, Osaka University, Nakanoshima, Osaka 530, Japan. 0379-0738/86/$03.50
0 1986 Elsevier Scientific Publishers Ireland Ltd. Printed and Published in Ireland
chronic poisoning of selenium in animals was first known in the 1930’s when they grazed on soils that contain large amounts of selenium, while only an industrial problem has been posed for man [ 11. “Blind staggers” and “alkali disease” are known as acute and chronic poisoning of animals, respectively. The former is characterized by defective vision and the latter by general wasting away of the body with many symptoms. Although there are many reports on selenium poisoning in animals, reports in man are rare [ 1,6]. In this report, we describe an autopsy case of acute selenium poisoning, in which a 40-year-old female ingested “Gun Blue” for suicide, and selenium levels were determined in the tissues of the case and normal individuals. The concentrations of selenium in various human tissues are needed to evaluate chronic selenium poisoning in man and also the nutritional and functional significances of selenium in human tissues. Materials and methods For the determinations of selenium and some other trace metals, various tissues were dissected and stored at -20” C until analysis. Tissues of patients who died by violence and in whom no diseases were found were used for control (nine males and six females, aged 40-59 years old). Determination of selenium and heavy metals Heavy metals were determined by an atomic absorption spectrometer (Hitachi 208) with the exception of mercury which was determined by a Zeeman effect mercury analyzer (Hitachi 501). Selenium was determined by gas chromatography after the conversion into 5-chloropiaselenol by 4-chloroo-phenylenediamine in acidic solution by the method of Nakanishi and Toei [ 71 with modifications [8]. After hydrolysis of tissues (approx. 1 g) with 5 ml of a mixture of nitric acid and perchloric acid (3: 2) and 1 ml of sulfuric acid at 180°C for about 2 h, samples were adjusted to pH 1 using ammonium hydroxide, diluted to 10 ml with distilled water and washed twice with ethyl acetate. 5Chloropiaselenol was synthesized by heating samples in a boiling water bath for 5 min after additions of 5 ml of 0.01 M EDTA and 2 ml of 0.5% 4-chloro+phenylenediamine and then extracted with 2 ml of benzene. Authentic li-chloropiaselenol was also synthesized as described above from selenium dioxide. Samples were subjected to gas chromatograph equipped with an electron-capture detector and a glass column (0.3 X 200 cm) packed with 4% OV-17 Uniport HPS (Gasukuro Kogyo Inc.). The column and detector temperatures were maintained at 190°C to obtain a convenient separation. The limit of determination of selenium by the method was 0.02 pg and the recovery of added selenium (0.1-0.5 pg) to various tissues was 98%. 5-Chloropiaselenol derived from stomach contents was identified by a gas chromatograph-mass spectrometer (JMS-D300).
89
Case report A 40-year-old female was admitted to the hospital an hour after ingestion of a toxic substance in a suicide attempt. An almost empty bottle (90 ml) labeled “Super Blue (Gun Blue)” (4% H,Se03 and 2.5% CuS04 in 1.7 N HCl) was found in her room. She had severe vomiting of dirty, dark green-brown liquid with a nasty smell, diarrhea and delusional stupor. She was also violently confused and restless. Her heart rate was 120 bpm and the blood pressure was 102/62 mmHg at admission and decreased to 70/50 mmHg 4 h after admission. She complained sometimes of severe burning in her chest and abdomen. She became anuric and her condition progressively deteriorated until she went into cardiac arrest 8 h after ingestion of “Super Blue”. Clinical records of the patient were very poor and no laboratory test was done. Autopsy
findings
The patient’s height was 160 cm and her weight was 40 kg. Post-mortem examination revealed pulmonary edema with pleural effusion (200 ml each) and congestion in the kidney on sectioning. Stomach mucosa was severely hemorrhagic and erosive and about 100 ml of dirty, dark brown liquid was found in the stomach. A general congestion was found as a circulatory disturbance. Histologically, the lung showed moderate edema, infiltration of macrophages and fibrinoid necrosis in the alveolar wall. Interstitial edema, necrosis of proximal tubules and dilatation of glomerulus were found in the kidney. The liver had no pathological change except congestion. Levels of selenium viduals
and copper
in tissues of the patient
and normal
indi-
Some heavy metals in the stomach contents were surveyed and we found 15, 0.4 and 0.24 ccg/g of Zn, Cd and Hg, respectively. No As, Pb, Mn and Cr was detected and no alcohol, cyanate, paraquat or methamphetamine was detected. Selenium in stomach contents was identified by gas chromatography-mass spectrometry after conversion to 5-chloropiaselenol. EI-mass spectrum of 5-chloropiaselenol derived from the stomach contents was in agreement with that of the authentic one (Fig. 1). Table 1 shows tissue levels of selenium and copper of the patient and normal subjects. Levels of selenium and copper in tissues of normal individuals we determined are similar to those reported by Schroeder et al. [3] and Sumino et al. [9], respectively. Selenium levels in tissues of the patient were found to be 9-90-fold higher than that of control levels, whereas concentrations of copper were about 2-fold compared to that of normal subjects. The highest
90 TABLE
1
CONCENTRATION OF SELENIUM AND NORMAL INDIVIDUALS
Blood Brain Heart Lung Liver Spleen Pancreas Kidney Stomach contents Muscle Testes Small intestine a From b Mean ’ From
AND
COPPER
IN TISSUES
Selenium
(t&g wet)
Toxic case
Normal (n = 15)
Normal a (n = 6)
0.09 0.13 0.15 0.14 0.36 0.26 0.20 0.87
o.2c 0.13 0.28 0.15 0.54 0.34 0.30 1.09
2.6 1.2 12.7 5.4 3.5 14.2 18.0
+ * c + + + f *
OF THE PATIENT
Comer
0.03b 0.06 0.06 0.06 0.09 0.11 0.10 0.25
Toxic case
6.1 3.7 14.5 2.4 4.6 40.0
(fig/g wt) Normal (n = 44)
4.5 3.9 1.4 6.3
? f * t
1.5b 0.9 0.3 3.8
1.5 ? 0.5 2.6 ? 0.6
0.24 0.30 0.21
Schroeder et al. [3]. + S.D. Yang et al. [2].
levels of selenium kidney.
in tissues
of the
patient
were
found
in the lung and
Discussion We have described autopsy findings of acute selenium (selenious acid) poisoning and normal and toxic levels of selenium in human tissues in this report. Carter [6] has reported a lethal case of acute selenium poisoning with vomiting, moaning, purposeless movements’ of body and limbs, comatose condition and a characteristic strong smell of garlic or rotten onions on the patient’s breath, in which a boy aged 3 years ingested “gun-blueing compound” that contained 1.81% selenious acid. Symptoms and signs in the present case were similar to that reported by Carter [6], disturbances of the gastrointestinal tract and central nervous system. We were unable to diagnose the case as selenium poisoning since symptoms and signs and autopsy findings were similar to that of acute poisoning of heavy metals although selenium is not a heavy metal. We have diagnosed the present case as an acute selenium poisoning because toxic levels of selenium were found in blood and tissues examined (Table 1). Shortridge et al. [lo] found 1.63.1 yg selenium/g tissue in the liver and kidney of cattle of acute selenium poisoning, however, the selenium level in tissues of acute poisonings in humans were not determined [6]. We found 12.7 and 14.2 clg selenium/g
91
%_
Stomach
+ 218
content
103 76 183 I
250
%.
m/e
’ 218
5-Chloropiaselenol v = Se -A io3
0\ dr
76
183 II 50
I 100
1 150
I ,I, 200
250
m/e
Fig. 1. EI-mass spectra of 5-chloropiaselenol and stomach contents after conversion with 4-chloro-o-phenylenediamine. Apparatus: JEOL JMSD300 Mass spectrometer fitted with electron ionization source and JMA-2000 data system. Column: glass tube (0.2 X 200 cm) packed 4% OV-17 uniport HPS. Carrier gas: helium, 0.8 kg/cm’. Temperature: injection port, 220°C; column oven, 190°C; separator, 200°C; ion source, 250°C. Ionization voltage, 24 eV. Ion multiplier voltage, 1.9 kV. Switching rate, 2.3 s. Source pressure, 0.3 x lo-’ torr. Retention time, 2.5 min.
tissue in the lung and kidney of the patient, respectively, and 1.2-5.4 pg selenium/g tissue in various other tissues examined (Table 1). If it is supposed that the patient had ingested the total volume of a bottle of “Super Blue” (90 ml), we calculated that she ingested 90 mg selenious acid/kg. It is clear that this is far beyond LlX 5,, reported in animals (approx. 5 mg selenious acid/kg) [ 1,6]. A lethal dose of cupric sulfate administered orally
92
to a rat is known to be about 960 mg/kg [12] and the patient’s ingestion was estimated to be a maximum of 60 mg/kg. Autopsy and histological findings in the present case were the congestion, edema, hemorrhage in tissues and dilatation of capillaries of the lung and kidney. These were in agreement with previous reports [6,11]. We also determined normal levels of selenium in human tissues since very few data are available and found that less than 1 pg/g tissue in tissues examined which coincided with the report of Schroeder et al. [3]. The highest amounts were found in the kidney and then in the liver and spleen. Acknowledgement HK thanks Marc F. Wathen, advice.
Lecturer,
Osaka YMCA college,
for linguistic
References 1 C.G. Wilber, Toxicology of selenium: A review. Clin. Toxicol., 17 (1980) 171~-230. 2 G. Yang, J. Chen, Z. Wen, K. Ge, L. Zhu, C. Chen and X. Chen, The role of selenium in Keshan disease. Adu. Nutr. Res., 6 (1984) 203-231. 3 H.A. Schroeder, D.V. Frost and J.J. Balassa, Essential trace metals in man: Selenium.
J. Chronic Dis., 23 (1970) 4 Y.C. Awasthi, E. Beutler 5 6
7 8
9
10 11 12
227-243.
and S.K. Srivastova, Purification and properties of human erythrocytes glutathione peroxidase. J. Biol. Chem., 250 (1975) 5144-5149. R.J. Shamberger and C.E. Willis, Epidemiological studies on selenium and heart 578 (abstract). disease. Fed. Proc., 35 (1976) R.F. Carter, Acute selenium poisoning. Med. J. Aust., 1 (1966) 525-528. S. Nakashima and K. Toei, Determination of ultramicro amounts of selenium by gas chromatography. Talanta, 15 (1968) 1475-1476. K. Nakamuro, Y. Sayato, M. Tonomura and Y. Ose, Studies on selenium related compounds. I. The determination of selenium by gas chromatography. J. Hyg. Chem., 18 (1972) 237-242 (in Japanese). K. Sumino, K. Hayakawa, T. Shibata and S. Kitamura, Heavy metals in normal Japanese tissues. Amounts of 15 heavy metals in 30 subjects. Arch. Environ. Health, 30 (1975) 487-494. E.H. Shortridge, P.J. O’Hara and P.M. Marshall, Acute selenium poisoning in cattle. N. 2. Vet. J., 19 (1971) 47-50. J.H. Draize and O.A. Beath, Observations on the pathology of “blind staggers” and “alkali disease.” J. Am. Vet. Med. Assoc., 86 (1935) 753-763. H.F. Smyth, C.P. Carpenter, C.S. Weil, U.C. Striegel and J.S. Nycum, Range-finding toxicity data. VII. Am. Ind. Hyg. Assoc. J., 30 (1969) 470-476.
Forensic Science International, 31 (1986) 87-9 2 Elsevier Scientific Publishers Ireland Ltd.
AN AUTOPSY CASE OF POISONING AND SELENIUM
ACUTE LEVELS
SELENIUM (SELENIOUS IN HUMAN TISSUES
RYOJI MATOBA, HIROSHI KIMURA, EIKOH UCHIMA*, YAMADAa, YOICHI MITSUKUNIa and ICHIRO SHIKATA
TAKASHI
Department of Legal Medicine, Faculty of Medicine, Osaka Medical Examiner’s Office, Nakanoshima, qsaka 530 (Japan)
ACID)
ABE, TADASHI
University
and
“Osaka
(Received November 4, 1985) (Revision received December 23, 1985) (Accepted January 17, 1986)
Summary A case of fatal suicidal ingestion of “Super Blue (Gun Blue)” (gun-blueing) is presented. Post-mortem examination of the patient revealed pulmonary edema with pleural effusion and congestion of the kidney. Necrosis of proximal tubules was found in the kidney by histological examination. “Super Blue” contains 4% selenious acid and 2.5% cupric sulfate in HCl. Levels of selenium and copper in tissues of the toxic case and normal individuals were determined. The selenium levels of tissues of the patient were 9 -90-fold higher than that of normal subjects, whereas concentrations of copper were about 2-fold compared to that of control levels. The highest levels of selenium in the tissues of the patient were found in the lung, kidney and stomach contents. Key
words:
Selenium poisoning; Tissue selenium; Autopsy
Introduction Selenium is one of the essential trace elements in human and animal nutrition [l-3] and is known as an integral part of the enzyme of glutathione peroxidase of human erythrocytes [4]. It is known that 0.04 -0.1 ppm of selenium is required in the diet for health, while a deficient or toxic effect appears if the diet contains low selenium or more than 4 ppm, respectively [l]. Selenium deficiency is known as “white muscle disease” for animals and Keshan disease, an endemic cardiomyopathy in China, for humans [2]. Some relationships between selenium deficiency and human cardiovascular diseases have been suggested from epidemiological studies [5]. Acute and *Present address: Department of Legal Medicine, School of Medicine, University of the Ryukyus, Nishihara, Okinawa 90301, Japan. Address all correspondence to: Dr. Ryoji Matoba, Department of Legal Medicine, Faculty of Medicine, Osaka University, Nakanoshima, Osaka 530, Japan. 0379-0738/86/$03.50
0 1986 Elsevier Scientific Publishers Ireland Ltd. Printed and Published in Ireland
chronic poisoning of selenium in animals was first known in the 1930’s when they grazed on soils that contain large amounts of selenium, while only an industrial problem has been posed for man [ 11. “Blind staggers” and “alkali disease” are known as acute and chronic poisoning of animals, respectively. The former is characterized by defective vision and the latter by general wasting away of the body with many symptoms. Although there are many reports on selenium poisoning in animals, reports in man are rare [ 1,6]. In this report, we describe an autopsy case of acute selenium poisoning, in which a 40-year-old female ingested “Gun Blue” for suicide, and selenium levels were determined in the tissues of the case and normal individuals. The concentrations of selenium in various human tissues are needed to evaluate chronic selenium poisoning in man and also the nutritional and functional significances of selenium in human tissues. Materials and methods For the determinations of selenium and some other trace metals, various tissues were dissected and stored at -20” C until analysis. Tissues of patients who died by violence and in whom no diseases were found were used for control (nine males and six females, aged 40-59 years old). Determination of selenium and heavy metals Heavy metals were determined by an atomic absorption spectrometer (Hitachi 208) with the exception of mercury which was determined by a Zeeman effect mercury analyzer (Hitachi 501). Selenium was determined by gas chromatography after the conversion into 5-chloropiaselenol by 4-chloroo-phenylenediamine in acidic solution by the method of Nakanishi and Toei [ 71 with modifications [8]. After hydrolysis of tissues (approx. 1 g) with 5 ml of a mixture of nitric acid and perchloric acid (3: 2) and 1 ml of sulfuric acid at 180°C for about 2 h, samples were adjusted to pH 1 using ammonium hydroxide, diluted to 10 ml with distilled water and washed twice with ethyl acetate. 5Chloropiaselenol was synthesized by heating samples in a boiling water bath for 5 min after additions of 5 ml of 0.01 M EDTA and 2 ml of 0.5% 4-chloro+phenylenediamine and then extracted with 2 ml of benzene. Authentic li-chloropiaselenol was also synthesized as described above from selenium dioxide. Samples were subjected to gas chromatograph equipped with an electron-capture detector and a glass column (0.3 X 200 cm) packed with 4% OV-17 Uniport HPS (Gasukuro Kogyo Inc.). The column and detector temperatures were maintained at 190°C to obtain a convenient separation. The limit of determination of selenium by the method was 0.02 pg and the recovery of added selenium (0.1-0.5 pg) to various tissues was 98%. 5-Chloropiaselenol derived from stomach contents was identified by a gas chromatograph-mass spectrometer (JMS-D300).
89
Case report A 40-year-old female was admitted to the hospital an hour after ingestion of a toxic substance in a suicide attempt. An almost empty bottle (90 ml) labeled “Super Blue (Gun Blue)” (4% H,Se03 and 2.5% CuS04 in 1.7 N HCl) was found in her room. She had severe vomiting of dirty, dark green-brown liquid with a nasty smell, diarrhea and delusional stupor. She was also violently confused and restless. Her heart rate was 120 bpm and the blood pressure was 102/62 mmHg at admission and decreased to 70/50 mmHg 4 h after admission. She complained sometimes of severe burning in her chest and abdomen. She became anuric and her condition progressively deteriorated until she went into cardiac arrest 8 h after ingestion of “Super Blue”. Clinical records of the patient were very poor and no laboratory test was done. Autopsy
findings
The patient’s height was 160 cm and her weight was 40 kg. Post-mortem examination revealed pulmonary edema with pleural effusion (200 ml each) and congestion in the kidney on sectioning. Stomach mucosa was severely hemorrhagic and erosive and about 100 ml of dirty, dark brown liquid was found in the stomach. A general congestion was found as a circulatory disturbance. Histologically, the lung showed moderate edema, infiltration of macrophages and fibrinoid necrosis in the alveolar wall. Interstitial edema, necrosis of proximal tubules and dilatation of glomerulus were found in the kidney. The liver had no pathological change except congestion. Levels of selenium viduals
and copper
in tissues of the patient
and normal
indi-
Some heavy metals in the stomach contents were surveyed and we found 15, 0.4 and 0.24 ccg/g of Zn, Cd and Hg, respectively. No As, Pb, Mn and Cr was detected and no alcohol, cyanate, paraquat or methamphetamine was detected. Selenium in stomach contents was identified by gas chromatography-mass spectrometry after conversion to 5-chloropiaselenol. EI-mass spectrum of 5-chloropiaselenol derived from the stomach contents was in agreement with that of the authentic one (Fig. 1). Table 1 shows tissue levels of selenium and copper of the patient and normal subjects. Levels of selenium and copper in tissues of normal individuals we determined are similar to those reported by Schroeder et al. [3] and Sumino et al. [9], respectively. Selenium levels in tissues of the patient were found to be 9-90-fold higher than that of control levels, whereas concentrations of copper were about 2-fold compared to that of normal subjects. The highest
90 TABLE
1
CONCENTRATION OF SELENIUM AND NORMAL INDIVIDUALS
Blood Brain Heart Lung Liver Spleen Pancreas Kidney Stomach contents Muscle Testes Small intestine a From b Mean ’ From
AND
COPPER
IN TISSUES
Selenium
(t&g wet)
Toxic case
Normal (n = 15)
Normal a (n = 6)
0.09 0.13 0.15 0.14 0.36 0.26 0.20 0.87
o.2c 0.13 0.28 0.15 0.54 0.34 0.30 1.09
2.6 1.2 12.7 5.4 3.5 14.2 18.0
+ * c + + + f *
OF THE PATIENT
Comer
0.03b 0.06 0.06 0.06 0.09 0.11 0.10 0.25
Toxic case
6.1 3.7 14.5 2.4 4.6 40.0
(fig/g wt) Normal (n = 44)
4.5 3.9 1.4 6.3
? f * t
1.5b 0.9 0.3 3.8
1.5 ? 0.5 2.6 ? 0.6
0.24 0.30 0.21
Schroeder et al. [3]. + S.D. Yang et al. [2].
levels of selenium kidney.
in tissues
of the
patient
were
found
in the lung and
Discussion We have described autopsy findings of acute selenium (selenious acid) poisoning and normal and toxic levels of selenium in human tissues in this report. Carter [6] has reported a lethal case of acute selenium poisoning with vomiting, moaning, purposeless movements’ of body and limbs, comatose condition and a characteristic strong smell of garlic or rotten onions on the patient’s breath, in which a boy aged 3 years ingested “gun-blueing compound” that contained 1.81% selenious acid. Symptoms and signs in the present case were similar to that reported by Carter [6], disturbances of the gastrointestinal tract and central nervous system. We were unable to diagnose the case as selenium poisoning since symptoms and signs and autopsy findings were similar to that of acute poisoning of heavy metals although selenium is not a heavy metal. We have diagnosed the present case as an acute selenium poisoning because toxic levels of selenium were found in blood and tissues examined (Table 1). Shortridge et al. [lo] found 1.63.1 yg selenium/g tissue in the liver and kidney of cattle of acute selenium poisoning, however, the selenium level in tissues of acute poisonings in humans were not determined [6]. We found 12.7 and 14.2 clg selenium/g
91
%_
Stomach
+ 218
content
103 76 183 I
250
%.
m/e
’ 218
5-Chloropiaselenol v = Se -A io3
0\ dr
76
183 II 50
I 100
1 150
I ,I, 200
250
m/e
Fig. 1. EI-mass spectra of 5-chloropiaselenol and stomach contents after conversion with 4-chloro-o-phenylenediamine. Apparatus: JEOL JMSD300 Mass spectrometer fitted with electron ionization source and JMA-2000 data system. Column: glass tube (0.2 X 200 cm) packed 4% OV-17 uniport HPS. Carrier gas: helium, 0.8 kg/cm’. Temperature: injection port, 220°C; column oven, 190°C; separator, 200°C; ion source, 250°C. Ionization voltage, 24 eV. Ion multiplier voltage, 1.9 kV. Switching rate, 2.3 s. Source pressure, 0.3 x lo-’ torr. Retention time, 2.5 min.
tissue in the lung and kidney of the patient, respectively, and 1.2-5.4 pg selenium/g tissue in various other tissues examined (Table 1). If it is supposed that the patient had ingested the total volume of a bottle of “Super Blue” (90 ml), we calculated that she ingested 90 mg selenious acid/kg. It is clear that this is far beyond LlX 5,, reported in animals (approx. 5 mg selenious acid/kg) [ 1,6]. A lethal dose of cupric sulfate administered orally
92
to a rat is known to be about 960 mg/kg [12] and the patient’s ingestion was estimated to be a maximum of 60 mg/kg. Autopsy and histological findings in the present case were the congestion, edema, hemorrhage in tissues and dilatation of capillaries of the lung and kidney. These were in agreement with previous reports [6,11]. We also determined normal levels of selenium in human tissues since very few data are available and found that less than 1 pg/g tissue in tissues examined which coincided with the report of Schroeder et al. [3]. The highest amounts were found in the kidney and then in the liver and spleen. Acknowledgement HK thanks Marc F. Wathen, advice.
Lecturer,
Osaka YMCA college,
for linguistic
References 1 C.G. Wilber, Toxicology of selenium: A review. Clin. Toxicol., 17 (1980) 171~-230. 2 G. Yang, J. Chen, Z. Wen, K. Ge, L. Zhu, C. Chen and X. Chen, The role of selenium in Keshan disease. Adu. Nutr. Res., 6 (1984) 203-231. 3 H.A. Schroeder, D.V. Frost and J.J. Balassa, Essential trace metals in man: Selenium.
J. Chronic Dis., 23 (1970) 4 Y.C. Awasthi, E. Beutler 5 6
7 8
9
10 11 12
227-243.
and S.K. Srivastova, Purification and properties of human erythrocytes glutathione peroxidase. J. Biol. Chem., 250 (1975) 5144-5149. R.J. Shamberger and C.E. Willis, Epidemiological studies on selenium and heart 578 (abstract). disease. Fed. Proc., 35 (1976) R.F. Carter, Acute selenium poisoning. Med. J. Aust., 1 (1966) 525-528. S. Nakashima and K. Toei, Determination of ultramicro amounts of selenium by gas chromatography. Talanta, 15 (1968) 1475-1476. K. Nakamuro, Y. Sayato, M. Tonomura and Y. Ose, Studies on selenium related compounds. I. The determination of selenium by gas chromatography. J. Hyg. Chem., 18 (1972) 237-242 (in Japanese). K. Sumino, K. Hayakawa, T. Shibata and S. Kitamura, Heavy metals in normal Japanese tissues. Amounts of 15 heavy metals in 30 subjects. Arch. Environ. Health, 30 (1975) 487-494. E.H. Shortridge, P.J. O’Hara and P.M. Marshall, Acute selenium poisoning in cattle. N. 2. Vet. J., 19 (1971) 47-50. J.H. Draize and O.A. Beath, Observations on the pathology of “blind staggers” and “alkali disease.” J. Am. Vet. Med. Assoc., 86 (1935) 753-763. H.F. Smyth, C.P. Carpenter, C.S. Weil, U.C. Striegel and J.S. Nycum, Range-finding toxicity data. VII. Am. Ind. Hyg. Assoc. J., 30 (1969) 470-476.