??????? ???pa?, BPharm, BChem, BBA, PhD

1
??T????? ????????G??S

• ??????? ???pa?, BPharm, BChem, BBA, PhD
• ??ap????t?? ?a????t?? Fa?µa?e?t????

2
??t??e?µe?? ?a? S??p?? t?? ??????????a?

• ? ??????????a ap?te?e? t?? ep?st?µ? p??
  a?t??e?µe?? t?? ??e? t?? ep?ß?aße?? d??se?? p??
  e?as???? ??s?e? d?af???? d?µ?? ?a? p????e?s??
  (xenobiotics) st??? ???a??sµ??? ?a? ?d?a?te?a
  st?? ?????p?.
• ?et? t? 1900 s??e???e? ?a a?apt?sseta? ?a? ?a
  eµp???t??eta? µe s??????e? ???se?? ?a? µe??d???
  ??a t?? ?ata??af? t?? t?????? ep?d??se??, t??
  a????e?s? (ta?t?p???s? ?a? p?s?t??? p??sd????sµ?)
  t?? d???t????? ?a? t? ?e?ape?a t?? d???t????se??.

3
??t??e?µe?? ?a? S??p?? t?? ??????????a?

• ? ??????????a µe?et? t??? µ??a??sµ??? d??s??
  t?????? ??s??? p?? p???a???? ??e?e? ? ?????e?
  d???t????se?? ?a??? ?a? t?? a????e?s?,
  ta?t?p???s? ?a? ap?t?µ?s? t?? ???d???? p??
  ap??????? ap? t? ???s? d?af???? ??s??? p?? µp??e?
  ?a p???a??s??? t?????t?ta.
• ? ??????? ??????????a as???e?ta? µe t?? ?ata??af?
  t?? s?µpt?µ?t?? ap? t?? t????? d??s? d?af????
  ??s??? ?a? t?? a??pt??? a?t?d?t?? ?a?
  ?e?ape?t???? s??µ?t?? ????? ?a? ???????
  d???t????se??.

4
?st????? ???????

• ???f??a d???t???a f?t???? ? ?????? p????e?s??
  ???s?µ?p??????a? ??a t? ??????, p???µ???,
  d???f???e?, a?t??t???e?.
• ??p???? Ebers (?1500 p.?.) ?e???aµß??e?
  p????f???e? ??a d??f??a d???t???a, p.?. a????t?,
  ?p??, ßa??a µ?ta??a (µ???ßd??, ?a????). ???µ? ??a
  t? da?t???t?da, a??a??e?d? Atropa, ?.?.
• ?pp????t?? (?400 p.?.) ?a????µ?se ta d???t???a
  ?a? ??ese t?? ß?se?? t?? ???????? ??????????a?.

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?st????? ???????

• Te?f?ast?? (370-286 p.?.) ?a??t?? t??
  ???st?t???, a?af??eta? se d???t????d? f?t? st?
  ???? t?? De Historica Plantarum.
• ???s?????d?? ?a????µ?se ta d???t???a a?????a µe
  t?? p????e?s? t??? (f?t????, ??????, ????t??
  p????e?s??).
• S????t?? ????f??????e µe t? ???e??.
• ?????d?t?? ?p????se ta a?t?d?ta (?????dat?sµ??).

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?st????? ???????

• ?a???e?s?? (1493-1541) ?????e? ? ?p?st?µ?????
  a?a??t?s? ?a? te?e???e? ? µ?st???sµ?? ap? ta
  ?????a t?? ???st?t???. ?p?st????e ?t? ?
  d???t????d?? d??s? ?fe??eta? st?? pa???s?a µ?a?
  ??µ???? ??s?a? p?? d?a t?????. ?d?a??se t??
  ep?st?µ????? s???? ?a? t?? pe??aµat?sµ? ??a t??
  e??tas? t?? t?????? ep?d??se?? ?a? d?a????se t??
  t?????? ap? t?? ?e?ape?t???? ?d??t?te? t??
  ??µ???? ??s???.
• ?p?st????e ?t? ?? t?????? ??s?e? pa???s??????
  e?e?d??e?µ??? d??s?, ?e????? p?? a???te?a ?d???se
  st?? ep????s? t?? ?e?ape?t???? de??t? ?a? t??
  d?s?-e?a?t?µe?e? d??se?? t?? t?????? ??s???.

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?st????? ???????

• Orfila ?spa??? ??at??? st? Ga???a. ? p??t??
  t?????????? p?? ???s?µ?p???se ta de??µata
  a?t???a? ?a? t? ??µ??? a????s? ?? ap?de??t???
  st???e?a ??a t? d?????s? d???t????se??. Te??e?ta?
  ? ?eµe???t?? t?? ?at??d??ast???? ??????????a?.
• Claude Bernard ?e??t?se t? ???????? ?a? s???ßa?e
  st?? a??pt??? t?? ??????????a? ?a? t??
  Fa?µa??????a?.

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S??????? ??????????a

• ??apt?????e ta te?e?ta?a 100 ?????a. ????se st?
  d????e?a t?? 2?? pa???sµ??? p???µ?? µe t? s???es?
  d?af???? fa?µ????, e?t?µ??t????, p??eµ???? ae????
  ?a? ß??µ??a????? ??s???.
• ?? Curie ?a? Becquerel a?a?????a? t?? ?ad?e?e????
  ??s?e? (t??????). ? a?a?????? t?? ß?taµ????
  ?d???se st?? a??pt??? µe??d?? ??a t?? p?s?t???
  t??? p??sd????sµ? (??a ap?f??? t?????t?ta? ????
  ?pe?d?s?????a?).

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S??????? ??????????a

• ?et? t? 1920 ????se ? e?tat??? µe??t? t?? e??se??
  a?se????? (?at? t?? s?f????) ?? ?p??e? p???a????
  ??e?e? ?a? ?????e? d???t????se??.
• ? a?a?????? t?? d??????d?fa?????t???????a??a????,
  (DDT - e?t?µ??t???) p?????se t?? t?????????a t??
  pa?as?t??t???? fa?µ????.
• ? a?a?????? t?? s???f??aµ?d??? (p?????esa?
  ?a??t??? ap? ?ef???? a?ep???e?a) ep?ßa?e t???
  t????????????? e??????? st?? fa?µa?e?t???? ??s?e?
  ?a? ?d???se st?? ?d??s? t?? FDA (1938).

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S??????? ??????????a

• ?eta?? 1940 ?a? 1946 a?a?a??f???a? d??f??a
  ???a??f?sf????? e?t?µ??t??a ?a? pa?as?t??t??a.
• Sta p?a?s?a t?? ??????????a? ?a? t??
  Fa?µa??????a? a?apt?????a? ?? ß?se?? t?? ??µ????
  ?a????????es?? ?a? t?? µetaß?????? e?e???p???s??
  t?? ??µ???? ?a??????????
• ?et? t? 2? pa???sµ?? p??eµ? ????se ? ??sp?s?
  ?a????? asfa?e?a? ??a t?? ?ata????s? t?? t??f?µ??
  ?a? t?? fa?µ???? ?a? ? ??e???? t?????t?ta? ??a
  p???? p?????ta. ? ?ata??af? ???sµ????
  epa??e?µat???? ??s?? ?d???se st?? a??pt??? t??
  ?p?d?µ????????? ??????????a?.

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S??????? ??????????a

• ?? de?aet?a t?? 1960, ? pe??pt?s? t?? ?a??d?µ?d??
  p?????ese te?at??e??se?? (f???µe??a) sta ?e????
  ?a? ?at?st?se a?a??a?a t?? t???????????
  a???????s? t?? fa?µ???? p?? ep??e????? t??
  eµß??????es? (te?at????a).
• ??te a?apt?????e ?a? ? ??a??t??? ??????????a, µe
  t?? a??pt??? a?a??t???? µe??d?? ??a t?? p???t???
  (ta?t?p???s?) ?a? p?s?t??? p??sd????sµ? t??
  t?????? ??s???.
• Bruce Ames ?? t?????? ??s?e? ß?????e ?t?
  p???a???? µeta????e?? µet? ap? µetaß?????
  e?e???p???s? ?a? a?t? ?d???se st?? a??pt???
  ???se?? ??a t? µeta??a??????es? ?a? t??
  ?a????????es?.

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S??????? ??????????a

• ?e?aet?a 1980 ??pa?s? t?? Love Canal ap?
  t????? ß??µ??a???? ap?ß??ta, ?d???se st??
  a??pt??? t?? ???µ??a????? ?a? ?e??ßa????t????
  ??????????a? p?? ap?s??p??? st? µe??t? t??
  ??pa?s?? t?? a??a, t?? ?d?t???? ?a? edaf????
  pe??ß?????t?? ap? t?????? ??s?e?, ??µ??? ap?ß??ta
  ?a? ß??µ??a???? ??µata.
• S?µe?a p?? ? ??pa?s? t?? pe??ß?????t?? ??e? ??ße?
  te??st?e? d?ast?se?? ?a? a??????e ? ?ata????s?
  t?? t??f?µ?? ?a? t?? fa?µ????, ? t????????????
  ??e???? e??a? apa?a?t?t?? ?a??? ?a? ? s?µas?a t??
  ??????????a?.

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??e?d??e?se?? ??????????a? (??µe??)

• ?e????af??? ??????????a ??et??e? t?? d???µas?e?
  p?? apa?t???ta? ??a t?? ap?t?µ?s? t?? ???d????
  ap? t?????? ??s?e? ?a? t? ??sp?s? ?a?????
  asf??e?a? ??a t? ???s? ??µ???? ??s???.
• ????a?? ??????????a ??et??e? t??? µ???a????
  µ??a??sµ??? d??s?? t?? t?????? ??s???. ? ???s?
  t?? µ??a??sµ?? d??s?? e??a? p???t?µ? ??a t?
  ?e?ape?a d???t????se?? ?a? t?? a??pt??? µe??d??
  a???????s?? t?? ???d???? ap? t?????? ??s?e?.

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??e?d??e?se?? ??????????a? (??µe??)

• ??????? ??????????a ?s???e?ta? µe 1)
  S?µpt?µat?????a d???t????se??, 2) ta µ?t?a
  ?e?ape?t???? a?t?µet?p?s??, 3) ta a?t?d?ta ?a? 4)
  t?? d?ad??as?e? ap?t?????s??.
• ?e??ßa????t??? ??????????a ?e?et? t?? ep?d??se??
  t?? t?????? ??s??? st? a????, ?d?t??? ?a? edaf???
  pe??ß?????.
• ????t?????????a ?s???e?ta? µe t?? ep?d?as?
  t?????? ??s??? se d??f??a ????s?st?µata.
  ??t??e?µe?? t?? ap?te???? ? µetaf???, µetaß?????
  t??? ?a? a????ep?d??se?? t?? t?????? ??s??? µe t?
  pe??ß?????.

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??e?d??e?se?? ??????????a? (??µe??)

• ??a??t??? ??????????a ?s???e?ta? µe t?? a??pt???
  ?a? efa?µ??? a?a??t???? µe??d?? ??a t?? ta?? ?a?
  a???p?st? p???t??? ?a? p?s?t??? p??sd????sµ? t??
  t?????? ??s??? se d??f??a ?p?st??µata.
• ?at??d??ast??? ??????????a ???????e ap? t??
  ??a??t??? ?a? ??????? ??????????a ?a? as???e?ta?
  µe t? d?????s? ?a?at?f???? d???t????se?? ?a? t??
  ?a????sµ? t?? s??????? ??t? ap? t?? ?p??e? ep???e
  ? ???at??.

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??????t?ta ????t?????

• ????t???? ?e??e?ta? ???e ??s?a p?? µp??e? ?a
  ep?f??e? ep?ß?aße?? ep?d??se?? ? ?a? ???at? se
  ??a? ???ta ???a??sµ?.
• ? t?????t?ta µ?a? ??s?a? e?a?t?ta? ap? t? d?s?
  p?? ?aµß??eta? ?a? s?ed?? ??e? ?? ??s?e? µp?????
  ?a e??a? d???t???a se s???e???µ??e? d?se??
  (d?s?e?a?t?µe?? t?????t?ta ?a???e?s??).
• ? LD50 (µ?s? ?a?at?f???? d?s?) ??a t? d??????
  e??a? 0.001 mg/kg s?µat??, e?? ??a ???e? ??s?e?
  e??a? p??? µe?a??te??, p.?. g/kg.

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??????t?ta ????t?????

• ? LD50 e??a? de??t?? µ??? t?? ??e?a? t?????t?ta?
  ?a? de? ??e? ???s? st?? ?????e? t?????t?te?.
  ?e????? ??s?e? se p?s?t?te? µ????te?e? t?? LD50
  µp????? ?a d???? sa? ?a????????a, te?at????a ?
  a??e???????a.
• ? t????? d??s? µ?a? ??s?a? e?a?t?ta? ap? t??
  p?s?t?t? t??, t?? ?d? e?s?d?? st?? ???a??sµ?, t?
  µetaf??? t?? st?? t?p? d??s?? (???a?a-st????), t?
  d????e?a d??s?? ?a? t?? e?a?s??s?a t?? ???a??sµ??
  st?? ??s?a.

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?d?? ??s?d?? ?a? ??p?? ???s??

• ?? t?????? ??s?e? e?s?????ta? ap? t? G?S (????a
  t? st?µa), d??µa, p?e?µ??e? ? pa?e?te????. ?
  pa?e?te???? e?s?d?? p???a?e? ta??te?? ?a?
  ap?te?esµat???te?? d??s?.
• ? t?????t?ta e?a?t?ta? ep?s?? ?a? ap? t? f???a
  µetaf???? t?? t?????? ??s?a?, p.?. a? ? f???a?
  e??a? ??p?f???? ? ?d??f???? p?? µp??e? ?a a???se?
  ? ?a d?e??????e? t?? ap????f?s? t?? t??????
  ??s?a?.

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?????e?a ?a? S????t?ta ???es??

• ? t?????t?ta e?a?t?ta? ??? µ??? ap? t? d????e?a
  ???es?? a??? ?a? ap? t? s????t?ta ???es?? st??
  t????? ??s?a.
• ??e?e? d???t????se?? pa?at?????ta? µ?sa se 24
  ??e? ? ?a? ????te?a s?????? ap? ef?pa? ??????s?
  t?? t?????? ??s?a?. ?????e? d???t????se?? µp??e?
  ?a eµfa??s???? µet? ap? ?µ??e?, µ??e? ? ?t?, µet?
  t?? ???es? t?? ???a??sµ?? st?? t????? ??s?a ???e
  ?µ??a, eßd?µ?da, µ??a ? ?t?.
• St?? ?????a t?????t?ta pa??e? ???? ? d????e?a ?a?
  s????t?ta ???es?? ?a??? ?a? ? ßa?µ?? s?ss??e?s??
  t?? t?????? ??s?a? ap? t?? ???a??sµ?.

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??d? ??????? ??e??e???

• ???e?????? ??t?d??se??
• ???s???????? a?t?d??se?? t?? ???a??sµ?? ????
  p??????µe??? e?a?s??t?p???s?? st?? t????? ??s?a
  (?pe?e?a?s??s?a). ?? d?s?-e?a?t?µe?e? a??e??????
  a?t?d??se?? µp??e? ?a p???a??s??? a??µ? ?a?
  ???at?.
• ?? a??s?p???t??? s?st?µa d?e?e??eta? ?a?
  pa?????ta? a?t?s?µata. ? e? ???? ???es? t??
  ???a??sµ?? st?? ?d?e? ??s?e? ?d??e? se a??e??????
  a?t?d??se?? ?pe?e?a?s??s?a? p?? µp????? ?a
  ?ata?????? ?a? se a?af??a?t??? s??.
• ?? a??e?????? a?t?d??se?? e?d??????ta? µe
  de?µat?t?da, e?????µa, ???sµ? ? e????µa µat???
  ?a? µe????? f???? µe ß?????spasµ?.

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??d? ??????? ??e??e???

• ??t?d??se?? ?d??s????as?a?
• ???µa?? a?t?d?as? t?? ???a??sµ?? se µ?a ??s?a p??
  ?a?????eta? ?e?et??? (p.?. ? s??????????????
  p???a?e? µ???a????s? ?a? ?p???a ??a a??et?? ??e?,
  ???? ?t?p?? ?e?d??????este??s??).
• ?µese? ?a? ?????e? ??????? ?p?d??se??
• ?.?. ? d?a?????st??ßest???? ?d??e? se ap?ß????
  eµß???? ?a? a??pt??? ?a?????? t?? µast??.

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??d? ??????? ??e??e???

• ??p???? ?a? S?st?µat???? ??????? ???se??
• ?.?. ?s???? ???a, ß?se??, ??e?d?t??? p???a????
  t?p???? d??se?? sta s?µe?a t?? s?µat?? µe ta
  ?p??a ?????ta? se epaf?.
• ? tet?aa?????????? µ???ßd?? e?t?? ap? t?p????
  d??se?? p???a?e? ?a? s?st?µat??? d??s? st? ??S
  ?a? t??? ?ef????.
• ? t????? d??s? µp??e? ?a e?d????eta? se ??a ?
  pe??ss?te?a ???a?a, p.?. ?pa?, ?ef???, µ?e??
  ?st?? (???a?a-st????).

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??d? ??????? ??e??e???

• ?????ep?d??se?? ??????? ??s??? ?p?? ?a? µe t??
  a????ep?d??se?? fa?µ????, ?? t?????? ??s?e?
  µp????? ?a a????ep?d??s??? µe f??µa?a, ???e?
  t?????? ??s?e? ? t??f?µa µe ap?t??esµa ?a
  ep??eas?e? ? ap????f?s? t???, ? ßa?µ?? s??des??
  t??? µe t?? p??te??e? t?? p??sµat??, ?
  µetaß???sµ?? t??? ?a? ? ap?????s? t???
  (fa?µa??????t???? a????ep?d??se??), µe ap?t??esµa
  t?? a???s? ? µe??s? t?? t?????t?t?? t???.
• ?p?????? ?a? ?? fa?µa??d??aµ???? a????ep?d??se??
  (?????s? ? µe??s? t?????? d??s?? ? s??????a).
  ?a??de??µa s??????a? ?s?? af??? t??
  ?pat?t?????t?ta ap?te?e? ? s??d?asµ?? a??a?????
  ?a? tet?a????????a?a. ?a?ade??µata e???det???s??
  (ap?t?????s??) t?? t?????? d??s?? ap?te???? ta
  d??f??a a?t?d?ta (e?d??? ? ?e????).

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??d? ??????? ??e??e???

• ????? st?? ??????? ??s?e?
• ?e????? f???? pa?at??e?ta? µe??µ??? e?a?s??s?a se
  µ?a t????? ??s?a, st?? ?p??a ta ?t?µa ?????
  e?te?e? ??a ??p??? ??????? d??st?µa (a??pt???
  a?t???? ? a?????, p.?. e?a?t?s?????e? ??s?e?).
  ??t? µp??e? ?a ?fe??eta? se µe??µ??? e?a?s??s?a
  t?? ?st?? ? t?? ???????, ? se a?epa???? p?s? t??
  t?????? ??s?a? p?? f???e? st?? t?p? d??s??.
  ?e????? f???? ? a?t??? µp??e? ?a ?fe??eta? st??
  epa???? p??te???? p?? desµe???? t? d???t????
  (p.?. ?? µeta????e?????e? desµe???? t? ??dµ?? ?a?
  t? ad?a??p?????).

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??d? ??????? ??e??e???

• ??s?-e?a?t?µe?? ??????t?ta
• ?as??? a??? t?? ??????????a? e??a? ?t? ? ßa?µ??
  t?????t?ta? e?a?t?ta? ap? t? d?s?. ?????? t??????
  ??s?e? pa???s?????? d?af??et??? ßa?µ? t?????t?ta?
  sta d??f??a ???a?a ? ?st???. ?e???? s?µas?a ??e?
  ? s?ss??e?s?, ? d??s? ?a? ? µetaß???sµ?? µ?a?
  t?????? ??s?a? se ??a s???e???µ??? ???a?? ? ?st?.

26
??s? Ta?at?f???? ??s? (LD50)

• ?p?te?e? t?? ef?pa? d?s? µ?a? t?????? ??s?a? p??
  p???a?e? ???at? st? 50 t?? pe??aµat?????.
  ?p?te?e? t?? p??t? pe??aµat??? d?ad??as?a t??
  t?????t?ta? µ?a? ??µ???? ??s?a?.
• G?a t?? p??sd????sµ? t?? ???????µe d?af??et????
  d?se?? t?? ??s?a? se ?µ?de? pe??aµat????? ?a?
  p??sd?????eta? ? ???s?µ?t?ta ??a ???e d?s?.
  ?????pte? µ?a s??µ?e?d?? ?aµp??? ? ?p??a st??
  ????a t?? ? ??e? t? ???s?µ?t?ta () ?a? st??
  ????a t?? ? t?? d?se?? (mg/kg) se ???a???µ???
  ???µa?a. ?a ???a t?? ?aµp???? a??????? ap? µ?d??
  (0) ???s?µ?t?ta ?a? f?????? t? 100 se p???
  ?????? d?se??.

27
???sd????sµ?? t?? DL50
28
??s? Ta?at?f???? ??s? (LD50)

• ? LD50 ap?te?e? t?? p??t? de??t? t?????t?ta? µ?a?
  ??s?a? ?a? ?e???? ?e??e?ta? s?µa?t??? st??
  a?t?µet?p?s? t?? d???t????se??. ??st???? ?µ??,
  sp???a ??e? p?a?t??? ??????? s?µas?a. ? LD50 de?
  ?d??e? p??t?te se µ?a sta?e?? t?µ? ???? t??
  d?af???? t?? e?a?s??s?a?, ?d??s????as?a? ?a? t??
  µetaß???sµ?? µeta?? t?? pe??aµat????? ?a? t??
  a????p??.
• Sp???a e??a? d??at? µ?a ??aµµ??? s?s??t?s? t??
  ded?µ???? t?? pe??aµat????? ?a? t?? a????p?? p??
  ?a ep?t??pe? µ?a a???p?st? p??ß??? t?? ded?µ????
  t?? pe??aµat????? st?? ?????p?. ?p???e? µ?a
  e????st? ap?te?esµat??? d?s? ??a ???e ??s?a p??a
  ap? t?? ?p??a a????e? ?a eµfa???eta? ? t?????
  d??s?.

29
??s? Ta?at?f???? ??s? (LD50)

• ? t?µ? t?? LD50 e?a?t?ta? ap? 1) ?d? e?s?d?? t??
  t?????? ??s?a? (d?af??et??? ß??d?a?es?µ?t?ta), 2)
  e?d?? pe??aµat?????, 3) f???, 4) f???, 5) ?????a,
  6) ß????, 7) d?a??t? st?? ?p??? d?a?????e ?
  ??s?a, 8) µetaß???sµ?, 9) ta?t?????? ???? ?????
  fa?µ???? ?a? 10) f?s??????a t?? pe??aµat?????.
  ?ts? ? LD50 ?a p??pe? ?a e?f???eta? µe ??p??a
  ???a. (95 ???a eµp?st?s????).
• ? p??sd????sµ?? t?? LD50 ???eta? s?µf??a µe t??
  µe??d??? t?? Litchfield ?a? Wilcoxon, ?a??? ?a?
  t?? Bliss ?a? Finney p?? apa?t??? stat?st???
  epe?e??as?a µe ?/? ?a? s?et??? µe???? a???µ?
  pe??aµat????? (40-50). S?µe?a ?p?????? µ???d??
  p?? ???s?µ?p????? ????te?a pe??aµat???a ?a?
  d????? a???p?sta ap?te??sµata.

30
??????t?ta ?a? Te?ape?t???? ?e??t??

• G?a ???e ??µ??? ??s?a (p.?. f??µa??) d?a??????µe
  d??f??e? d?se?? st?? ?p??e? p???a?e?ta? e?te
  ep???µ?t? ?e?ape?t??? d??s?, ? t?????t?ta ?
  ???at??. ??? ?????a t?? ?e?ape?t???? de??t?
  e?s??a?e ? Paul Ehrlich t? 1913.
• Te?ape?t???? de??t?? ????eta? ? ????? t?? µ?s??
  d?s?? t?? ??s?a? p?? apa?te?ta? ??a ?a p???a??se?
  t?????t?ta ? ???at? st? 50 t?? pe??aµat?????
  p??? t? µ?s? d?s? p?? apa?te?ta? ??a ?a ep?f??e?
  t? ep???µ?t? ?e?ape?t??? ap?t??esµa.

31
??????t?ta ?a? Te?ape?t???? ?e??t??

• Te?ape?t???? ?e??t?? TI LD50/ ED50 ? ?D50/?D50
  
• ?p?? ED50 ? µ?s? ap?te?esµat??? d?s? ?a? ?D50 ?
  µ?s? t????? d?s?. ?s? µe?a??te??? e??a? ?
  ?e?ape?t???? de??t?? t?s? µe?a??te?? e??a? t?
  e???? asfa?e?a? t?? t?????? ? fa?µa?e?t????
  ??s?a?.

32
???e?t??? ??????t?ta

• ?p?????? t?????? ??s?e? p?? p???a???? ?e???e?µ???
  t?????t?ta (se p???? ???a?a ?a? ?st???) ?a? ???e?
  p?? p???a???? t?????t?ta se ??a ???a?? ? ?st?.
  ?ts?, ?? t?????? ??s?e? ? ta f??µa?a eµfa??????
  e??e?t??? d??s? se ??p??? s???e???µ??? ???a?? ?
  ?st? ? ???a??sµ? (p.?. a?t?ß??t???). ? e??e?t???
  t?????t?ta µp??e? ?a ?fe??eta? se
  t?????????t????? ?????? (d?af???? st??
  ap????f?s?, ?ata??µ? e??e?t??? s?ss??e?s? se
  ??a ???a?? ? ?st? -, µetaß???sµ? ?a? ap?????s?) ?
  st?? e?e?d??e?µ??? d??s? t?? st? ???a??-st???.

33
???e?t??? ??????t?ta

• ? e??e?t??? t?????t?ta ??µ???? ??s??? p??
  pa?at??e?ta? µeta?? t?? d?af???? e?d?? s????
  ?fe??eta? se d?af???? p?? pa???s?????? ta ?????
  ?a? f?t??? ??tta?a ? µ????ß?a.

34
???a??sµ?? ???s?? ??????? ??s???

• ?? ??µ???? ??s?e? p???a???? t?????? d??se?? µ?s?
  µetaß???? p?? ep?f????? st? ß????µe?a ?a?
  f?s??????a t?? ???a??sµ??.

35
??t?d??se?? ??????? ??s???-?p?d?????
(Ste?e?e??e?t???t?ta)

• ?????? t?????? ??s?e? ? f??µa?a d???? µ?s?
  ?p?d????? se ?st???-st?????. ? a?t?d?as? µe t??
  ?p?d???a s?????? e??a? a?t?st?ept? ?a?
  pe?????feta? ap? t?? e??s?s? R T ? R.T. ?
  a?t?d?as? a?t? ?a?a?t????eta? ap? ??p??a
  s?????e?a Kd TR/RT. ? t????? ??s?a
  µp??e? ?a e??a? e?d??e??? ? e???e??? p??
  d?e?e??e? ? a?ast???e? µ?a f?s???????? ?e?t?????a
  t?? ???a??sµ??. S???? ?? t?????? ??s?e? d???? ??
  a?ast??e?? ?a? pa?eµp?d????? t? ?e?t?????a
  e???µ??. ?????? f???? ?? a?t?d??se?? t??????
  ??s??? ?a? ?p?d????? e??a? ste?e?e??e?t????.

36
??ast??? ?e?t?????a? ??tta????? ?eµß?a???

• ? d?at???s? t?? ??e?t????µ???? d??aµ???? ?a?
  ?e???? ? d?µ??? ?a? ?e?t??????? sta?e??t?ta t??
  ??tta????? µeµß?a??? ap?te???? ßas??? st???e?a
  st? f?s??????a t?? ?st??.
• ?e????? ??µ???? ??s?e? µp????? ?a ep??e?s??? t??
  ?at?stas? p???s?? t?? ??tta????? µeµß?a???. S????
  ep??e????? t? ??? ???t?? a?ast?????ta? t???
  d?a????? t??? (p.?. ? tet?ad?t????? d?a
  a?ast?????ta? t?? µetaf??? ???t?? Na ?a?
  p???a?e? ?e???t????? d??s? ep??e????ta? t??
  epa?ap???s? t?? µeµß?a???). ???sµ???? ???a?????
  d?a??te? ep??e????? t? ?e?st?t?ta t?? ??tta?????
  µeµß?????.

37
??ast??? ?a?a????? ??tta????? ?????e?a?

• ?????? t?????? ??s?e? a?ast?????? t?? ??e?d?s?
  t?? ?data??????? p?? ?d??e? st?? pa?a???? ATP. ?
  pa???? ???????? st??? ?st??? ep??e??eta? ap? ta
  ??a?????a, ta ??e?d?t??? d???t???a ?a? ta a??d?a.
  ?? ??t??fa????e? p???a???? e???t??s? t??
  e?e??e?a??? ap??eµ?t?? (ATP) (ap?s??e???
  ??e?d?t???? f?sf??????s?? ?a? a?ap?e?st????
  a??s?da?).

38
S??des? µe ???µ???a

• ???f??e? t?????? ??s?e? s??d???ta? µe ta e?e???
  ???t?a t?? e???µ?? p?? e??a? p??? ??t??? ??a t?
  µetaß???sµ? t?? ??tt????. ?.?. t? HCN s??d?eta?
  µe t? s?d??? t?? e???µ?? ??e?d?s? t??
  ??t????µat?? C, a?ast?????ta? ?ts? t? te????
  st?d?? st?? a??s?da µetaf???? ??e?t??????. ?? CO
  s??d?eta? ?s???? µe t?? a?µ?sfa????? ?a?
  s??µat??e? a???a???a?µ?sfa????? ?a? µe?????ta?
  ?ts? t?? µetaf??? ???????? st??? ?st???. ?????
  ßa??a µ?ta??a (Pb, Hg, Cd, As) s??d???ta? µe t??
  ?µ?de? SH d?af???? e???µ??. ???f??e? t??????
  ??s?e? s??µat????? e?e??e?e? ???e? µe ap?t??esµa
  t? ?????s? t?? ?st?? (??p?d??? ?pe???e?d?s?).

39
S??des? µe ???µ???a

• ?e????? t?????? ??s?e? d???? ?? ??e?t?????f??a
  ?a? a?t?d???? µe d??f??e? p?????f??e? ?µ?de?
  (-SH, -OH, -NH2) d?af???? e???µ?? ? p??te???? ?a?
  t?? ????ta?e????? (GSH), µe ap?t??esµa t??
  ?atast??? t?? d??s?? p????? ??t???? e???µ?? ? t??
  e???t??s? t?? ap??eµ?t?? t?? GSH ?a? ??e?d?t???
  stress.
• ?e????? t?????? ??s?e? e?s?????ta? st??
  e?d???tt???? ????, s?ss??e???ta? st?? p????a ?a?
  a?t?d???? ?µ???p????? µe p?????f??e? ??se?? st?
  DNA. ?.?. a??????s? t?? ???????? t?? 6-??s?? t??
  ???a????? ?d??e? se µeta??a??????es? ?a?
  ?a????????es? (??t???aµ??e?).

40
??ata?a??? t?? ?µ???stas?? t?? ?sßest???

• ?at? t? ?????s? t?? ??tt???? pa?at??e?ta? e?s?d??
  ???t?? asßest??? st? ??tta??. ??ata?a?? t??
  ?µ???stas?? t?? e?d???tt????? Ca µp??e? ?a
  p?????e? ap? d??f??e? t?????? ??s?e? p?? d????
  st? ep?ped? t?? ??tta????? µeµß????? ?a? t??
  µ??a??sµ?? µetaf???? asßest??? (??t??fa????e?,
  ?????e?, ?pe???e?d?a, d???????, µe???? µeta?????
  ???ta). ????s? t?? e?d???tta????? s?????t??s??
  t?? Ca ?d??e? se e?e???p???s? µ? ??ss?s?µ????
  e???µ?? ? d?e?e??e? e?d??????e?se? p?? p???a????
  d??spas? t?? DNA ?a? s?µp????s? t?? ???µat????.

41
???e?t??? ?atast??f? ??tt???? st??? ?st???

• ?e???e? d?se?? µa??a???? p???a???? e?f???sµ? t??
  ?t?paµ??e?????? ?e?????? sta ßas??? ??????a t??
  e??ef????. ?p?s?? s?ss??e?s? µe????? p?s?t?ta?
  ?ad?e?e???? ??d??? st? ???e?e?d? ad??a ep?f??e?
  e?f???sµ? t?? ??tt???? t?? ???e?e?d?.

42
??????t?ta ???? ?eta????e?? sta S?µat??? ??tta?a

• ??? pe??ss?te?e? f???? ?? t?????? ??s?e? p??
  a?t?d???? µe t? DNA p???a???? ?atast??f? t??
  ??tt????. ???sµ??e? f???? p???a???? ?e?et????
  a?????se?? (µeta????e??) p?? ?d????? se
  ?a????????es? (?e??t????? ?a????????a). ???et??
  µeta????e?? d????????ta? µ?s? t?? s?st?µat??
  ep?d?????se?? t?? DNA, ???sµ??e? ?µ??
  ???????µ???ta? ?a? ?d????? stad?a?? st?? a??pt???
  ?e?p?asµ?t??.

43
??????t?ta ???? ?eta????e?? sta S?µat??? ??tta?a

• ?p?st????eta? ?t? ta ?a????????a d?e?e?????
  (e?e???p?????) ta p??t?????????d?a ? p???a????
  µeta????e?? se ?????atasta?t??? ????d?a, µe
  ap?t??esµa t?? ??a??? ?a????????es?? (?e?????ta?
  ta p??ta st?d?a t?? a??pt???? ?e?p?as?a?).
• ?e????? ??s?e? de? p???a???? µeta????e?? ?µ??
  d?e?e????? t? ?e?p?as?a µet? t? st?d?? t??
  ??a????. ?? ??s?e? a?t?? d???? ?? p??a???e?? t??
  ?e?p?as?a? (a?ast?????? t? d?af???p???s? t??
  ??tt????) ? ?? µ?t????a (d?e?e????? t?? ??tta????
  ?????) ? a????ep?d???? µe t??? ?p?d??e?? t??
  a?apt???a??? pa?a???t??.

44
??e???? ??????t?ta? se ?e??aµat???a

• ???sd????sµ?? LD50
• ?p?te?e? d???µas?a ??e?a? t?????t?ta? se
  pe??aµat???a (p??t???a, a????a???, ? ???????a ?a?
  s?????), µe d??f????? ?d??? ??????s?? t?? t??????
  ??s?a?. ??d??ta? d??f??e? d?se?? ?a?
  p??sd?????eta? ? LD50 (mg/kg). ?p?s??
  ?ata???feta? ? ?e????te?? e????a t??
  pe??aµat????? (s?µpe??f???, ???a???? d?????s?,
  ???? t??f??). ?p?p???? s?µp?????eta? µe
  ?st?pa???????? e??tas? t?? ?e???t?µ???? e???µ?t??.

45
??e???? ??????t?ta? se ?e??aµat???a

• ??a ta pa?ap??? ap?s??p??? 1) St?? p??sd????sµ?
  t?? LD50, 2) ?a?a?t???sµ? ??????? sta ?p??a
  e?d????eta? ? ??e?a t?????t?ta, 3)
  a?ast?e??µ?t?ta ? µ? t?? t?????t?ta?, 4)
  p??sd????sµ? f?sµat?? d?se?? p?? p???a????
  t?????t?ta.
• ? t????? ??s?a µp??e? ?a efa?µ?s?e? ?a? st? d??µa
  (pa?aµ??e? ??a 24 ??e?) ?a? e?et??eta? ? pe?????
  t?? d??µat?? ??a t???? t?????t?ta ? µp??e? ?a
  d??e? ?p? µ??f? ae????µat?? ? s?st?µat?? ?ept??
  s????? ?p? d?asp???.

46
??????? ?p?d??se?? st? ???µa ?a? t??? ?f?a?µ???

• ???µa ?fa?µ??eta? s?????? st? d??µa ????e????.
  ?a?a???eta? ? pe????? t?? d??µat?? ap? t? t????µa
  ?a? efa?µ??eta? p?s?t?ta 0.5 ml ????? ? 0.5 g
  ste?e?? µe t? ß???e?a ???a? ??a t??????st?? 4
  ??e? (Draize test). ? ßa?µ?? t?????t?ta?
  e?t?µ?ta? ap? t?? ??tas? t?? e????µat??,
  ??d?µat??, d??ß??s?? ? ?e???? t?? ?a??p???s?? t??
  d??µat?? p?? p????????e.
• ?f?a?µ?? ? t????? ??s?a efa?µ??eta? ?p? µ??f?
  sta????? (0.1 ml) st??? ?f?a?µ??? ???????? ?a?
  e?et??eta? ? ?at?stas? t?? ?f?a?µ?? se d??f??a
  ??????? d?ast?µata µet? t? ??????s? t?? t??????
  ??s?a?.

47
????µas?a ?pe?e?a?s??s?a?

• ???s?µ?p?????ta? ??d??? ?????d?a. ? ??s?a
  ?????e?ta? e?te t?p??? st? d??µa ? e?d?de?µ??? µe
  ? ????? a??s?e??s??t??? ??a ?a a???se? t??
  ?pe?e?a?s??s?a.
• ? d???µas?a d?a??e? 2-4 eßd?µ?de? ?a? ???eta?
  e?d?de?µ??? ??????s? ?at? epa??????. ?et? ap? 2-3
  eßd?µ?de? ap? t? ??????s? t?? te?e?ta?a? d?s??,
  e?et??eta? t? d??µa ??a t? ßa?µ? t?? e????µat??
  p?? pa???s???e?.

48
????µas?a ??a ?p??e?a ??????t?ta µet? ap?
?pa?e???µµ??? ??????s?

• ?? d???µas?e? a?t?? ap?s??p??? st?? ?a????sµ? t??
  d?se?? p?? p???a???? ??? ??e?a a??? ?p??e?a
  t?????t?ta, ???????ta? ?at epa?????? t?? ??s?a.
• ?? s?????ste?? p??t?????? e??a? t? pa?a??t?
  ??????s? 3-4 d?af??et???? d?se?? t?? ??µ????
  ??s?a? se pe??aµat???a (a????a???) µ?s? t??
  t??f??. ?et? ap? 2 eßd?µ?de? a??????e?
  ???????e??ast???a?? ?a? ?st?pa???????? e??tas?
  t?? pe??aµat????? ??a t?? e?a???ß?s? t??
  t?????t?ta?.

49
????µas?a ??a ?p??????a ??????t?ta

• ?p??????a t?????t?ta e??a? a?t? p?? pa?at??e?ta?
  se ?????? µ??e? ap? t? ??????s? t?????? ??s?a? se
  pe??aµat???a (s?????? d?a??e? 90 ?µ??e?).
  S??pe?e? st?? ?a????sµ? d?se??, t?? ?ata??af?
  s?µpt?µ?t?? ?a? t? ?a?a?t???sµ? ??????? ?a? ?st??
  p?? ep??e????ta? ap? t?? epa?e???µµ??? ??????s?
  t?? t?????? ??s?a? (s?????? s?????).
• ????????ta? 3 d?af??et???? d?se??, µ?a ????? p??
  ?d??e? se ???at? t? 10 t?? pe??aµat?????, µ?a
  e?d??µes? ?a? µ?a p?? de? p???a?e? t?????t?ta. ?a
  pe??aµat???a e?et????ta? pe???d??? ??a t????
  t?????t?ta (ap??e?a ß?????, µetaß????
  ?a?d??a??e?a??? ?e?t?????a? ?a?
  s?µpe??f????-????t???t?ta?).

50
????µas?a ??a ?p??????a ??????t?ta

• ???µ? a?????????ta? ta ?e???t?µ??? e???µata ap?
  ta pe??aµat???a p?? apeß??sa? p??? ap? t? t????
  t?? d???µas?a? t?? 90 ?µe??? ? ???????e??ast???a??
  e??tas? a?µat??, ?????, d???µas?e? ?pat??
  (p??sd????sµ?? d?af???? e???µ??) pa??????
  p????f???e? ??a t?? t?????t?ta.
• St? t???? t?? d???µas?a? e?et????ta?
  ?st?pa???????? ??a ta pe??aµat???a ?a?
  ?aµß????ta? p????f???e? ??a t? ßa?µ? t?????t?ta?.
  ?? d???µas?e? t?? ??e?a? ?a? ?p??????a?
  t?????t?ta? p??pe? ?a ???????????ta? p??? ap? t??
  ??a??? t?? ???????? f?se?? st?? a??pt??? fa?µ????
  ??a a????p???.

51
??e???? ??a ?????a ??????t?ta

• ?? ?d?? ?p?? ?a? p??????µ???? ??a t?? ?p??????a
  t?????t?ta, a??? ??a µe?a??te?? ??????? d??st?µa
  (?????? µ??e? 2 ?t?). ?? ??????? d??st?µa
  e?a?t?ta? ap? t?? p??sd???µe?? ????? ??????s?? ?
  ???s?? t?? ??s?a? st?? ?????p?.
• ?????? ? a???????s? t?? ?????a? t?????t?ta? ??e?
  µe???? s?µas?a ??a t?? p??sd????sµ? t??
  t?????t?ta? p?? p???a?e?ta? ap? s?ss??e?s?
  ??µ???? ??s??? se pe??aµat???a.

52
????µas?a ??????t?ta? ?at? t?? ?µß??????es? ?a?
???pt???

• ???f??e? t?????? ??s?e? ep??e????? t??
  eµß??????es? ?a? t?? a??pt??? t?? ???a??sµ?? (ap?
  t? ?????s? µ???? ?a? t?? ef?ße?a). ???e? ??s?e?
  p???a???? t?????? e????e?e? sta ?e???t???
  ??tta?a. ?p?????? 4 in vivo d???µas?e? se
  pe??aµat???a ??a t? d???µas?a a?t?
  (???s?µ?p?????ta? a????a???).

53
????µas?a ??????t?ta? ?at? t?? ?µß??????es? ?a?
???pt???

• ????????ta? 2-3 d?af??et???? d?se?? t?? ??s?a?
  (20 pe??aµat???a a?? d?s?). ?a a?se????
  pe??aµat???a e?t??e?ta? st? ??µ??? ??s?a ??a 60
  ?µ??e?, e?? ta ?????? ??a 14 ?µ??e? p??? ap? t?
  d?asta???s? (????µ?p???s?). St? s????e?a
  p??sd?????eta? ? a???µ?? t?? pe??aµat????? p??
  ????µ?p??????a?, ? a???µ?? t?? ap?????? ?a? ?
  ???s?µ?t?ta, t? ß???? ?a? ?e???? ? a??pt??? ?a?
  ep?ß??s?, ? f?s??? ?at?stas? t?? ap?????? st?? 3
  p??te? eßd?µ?de?.

54
????µas?a ??????t?ta? ?at? t?? ?µß??????es? ?a?
???pt???

• G?a t?? a???????s? µ?a? ??µ???? ??s?a? ??a
  te?at????? d??s? ?? ??s?e? ????????ta? ?at? t??
  pe???d? t?? ???a?????es?? (aµ?s?? µet? t?
  ????µ?p???s?). ???s?µ?p?????ta? ???????a ?
  a????a??? (20 se ???e ?µ?da). ????????ta? 1-3
  d?af??et???? d?se?? (ap? 6? ??? 15? ?µ??a st???
  a????a???? ?a? ap? 6? ??? 18? ?µ??a sta
  ???????a).
• ?at?p?? ap?µa??????ta? ta ?µß??a (21? ?µ??a ??a
  a????a???? ?a? 31? ?µ??a ??a ta ???????a) ?a?
  e?et??eta? ? ???s?µ?t?ta ?a? ?? p??a??? a??µa??e?
  ?at? t?? a??pt???.

55
????µas?a ??????t?ta? ?at? t?? ?µß??????es? ?a?
???pt???

• ???tas? ??s??? p??? ?a? aµ?s?? µet? t?? t??et?
  ?????e?ta? ? ??s?a t?? 15? ?µ??a t?? ???se?? ?a?
  p??a? t?? t??et?? se a????a???? ?a?
  p??sd?????eta? ? ep?d?as? t?? st? ß????, t??
  ep?ß??s? ?a? a??pt??? t?? ?e????? ??a 3
  eßd?µ?de?.
• G?a t?? t?????t?ta st?? a?apa?a???? se µe????
  a???µ? pe??aµat????? ap? pe??aµat???a 1?? ?e?e??
  (F0) ?a? ap??????? 1?? (F1) ?a? de?te??? (F2)
  ???at????? ?e?e?? ???sd???????ta? 1) ? a???µ??
  t?? d?asta???se?? ?a? t?? pe??aµat????? p??
  ????µ?p??????a?, 2) ? a???µ?? t?? pe??aµat?????
  p?? ep?ß??sa?, 3) ? a???µ?? t?? ?e????? p??
  ep?ß??sa? aµ?s?? µet? t?? t??et?, 4) ta
  ?st?pa???????? ?a?a?t???st??? µe ?µfas? st?
  a?apa?a?????? s?st?µa t?? p??????? (F0) ?a? t??
  ap?????? d?? ?e?e?? (F1, F2).

56
????µas?a ??????t?ta? ?at? t?? ?µß??????es? ?a?
???pt???

• ???µ? e?t?? t?? in vivo d???µas??? a?apt?????a?
  ?a? in vitro d???µas?e? ??a t?? a???????s? t??
  t?????t?ta? st?? a?apa?a????, ?p?? ?a??????e?e?
  eµß???? ?a? ?ata???f??ta? ?? µetaß???? p??
  p???a???? ?? ??µ???? ??s?e? st? d?af???p???s? ?a?
  a??pt??? t?? eµß????.

57
????µas?a ??a ?eta??a??????es?

• ?e????? ??s?e? p???a???? a?????se?? (µeta????e??)
  t?? ?e?et???? ?????? (DNA) ?a? µe?????
  µeta????e?? µetaß?ß????ta? ?at? t?? ??tta????
  d?a??es? st??? ap???????. ?eta????e?? µp????? ?a
  s?µß??? sta ?e???t??? ??tta?a (?aµ?te? ?a? ????a)
  µe s?ßa??? s???pe?e? st??? ap??????? ? sta
  s?µat??? ??tta?a p?? µp????? ?a ?d???s??? st??
  a??pt??? ?e?p?as?a?. ?? ?? µeta????e?? s?µß???
  ?at? t? d????e?a t?? s??????? µp??e? ?a ?d???s???
  se ap?ß??? t?? eµß???? ? a??µa?? a??pt??? ?at?
  t?? eµß??????es?.

58
????µas?a ??a ?eta??a??????es?

• ????µas?e? a???????s?? t?? ??a??t?ta? t?? t??????
  ??s??? ?a p???a???? µeta????e?? 1) ? ?a???t???
  a????s? t?? ?e?et???? ?????? µe t? µ????s??p??
  µp??e? ?a a????e?se? s?ßa??? µeta????e?? ?a?
  a?????se?? ???µ?s?µ?t??. 2) ????µas?a se
  t???t??? ?????e?ta? µ?a ef?pa? d?s? t?? ??µ????
  ??s?a? se ??a a?se???? pe??aµat???? t? ?p??? st?
  s????e?a d?asta????eta? µe d?? ?????? ??a 8
  eßd?µ?de?. ?a ?????? pe??aµat???a ?a?at????ta?
  ?a? p??sd?????eta? ? a???µ?? t?? ???t?? eµß????
  p?? p????pt???.

59
????µas?a ??a ?eta??a??????es?

• Ames Test ???a? ? p?? d?µ?f???? d???µas?a
  µeta??a??????es??. ?e t? d???µas?a a?t?
  p??sd?????eta? a? ? ??µ??? ??s?a ? ?? µetaß???te?
  t?? p???a???? µeta????e?? se ??a st??e??? t??
  Salmonella typhimurium ap? t? ?p??? ?e?pe? t?
  ????µ? f?sf???ß?s???-ATP s???et?s?, t? ?p???
  apa?te?ta? ??a t? s???es? ?st?d????.

60
????µas?a ??a ?eta??a??????es?

• ?? st??e??? a?t? de? µp??e? ?a a?apt???e? se
  ??ept??? ????? ????? ?st?d???. ?µ?? µet? ap?
  µet???a?? (ap? t?????? ??s?e? ? t??? µetaß???te?
  t???) µp??e? ?a s??µat?se? ap????e?. St?
  d???µas?a a?t? ???s?µ?p?????ta? e?????sµata
  µ????s?µat?a??? ???sµat?? ?pat???tt????
  (pe??????? ????µa µetaß???sµ?? ?a? µetat??p??? ta
  ad?a?? µeta??a??????a se e?e????? µetaß???te?).
• Ta p??pe? ?a t???s?e? ?t? ? t???????????? ??e????
  ??µ???? ??s??? e??a? ?????ß???? ?a? ?d?a?te?a
  dapa?????.

61
?p?t?µ?s? ???d???? ap? t? ???s? ??????? ??s???

• ?p? p?a?t??? ?p??? ? ???s?µ?? pa?????ta? ??a t??
  eµf???s? t?? t?????t?ta? de? ap?te?e? ? ?d?a ?
  t?????t?ta t?? ??s?a?, a??? ? ???d???? p??
  s?et??eta? µe t? ???s? t?? ??s?a?. O? ???d????
  ????eta? ? p??a??t?ta ??a p?????s? ep?ß?aß??
  ep?d??se?? ap? ??p??a ??s?a se s???e???µ??e?
  s?????e?. ??t??eta, ?? asf??e?a ????eta? ?
  p??a??t?ta ?t? ? ??s?a de ?a p???a??se?
  ep?ß?aße?? ep?d??se??.
• ?ts?, p??? t?????? ??s?e? µp??e? ?a µ?? p???a????
  t?????? e????e?e? ?ta? de ???s?µ?p?????ta? ? ?ta?
  ???s?µ?p?????ta? ??t? ap? s?????e? asfa?e?a?. ?
  ßa?µ?? ep????d???t?ta? e?a?t?ta? ap? t?? s?????e?
  ?p? t?? ?p??e? ???s?µ?p??e?ta? µ?a t????? ??s?a.

62
?p?t?µ?s? ???d???? ap? t? ???s? ??????? ??s???

• ?at? t? d?ad??as?a a???????s?? t?? ???d????
  ?aµß????ta? ?p??? 1) ? ??a??t?ta t?? ??s?a? ?a
  p???a??se? t?????t?ta µe ß?s? ep?d?µ????????
  ded?µ??a. 2) ? p?????s? t?? t?????? ep?d??se??
  p?? µp??e? ?a p???a??se? µe ß?s? ta ap?te??sµata
  ap? pe??aµat???a. 3) ? ßa?µ?? ???es?? st?? t?????
  ??s?a. 4) ? ?a?a?t???sµ?? t?? ???d???? ??a
  ep?ß?aße?? d??se??.
• ? ap?t?µ?s? t?? ???d???? se pe??aµat???a µp??e?
  ?a ?a????se? ta asfa?? ep?peda ???s?? µ?a?
  t?????? ??s?a? ?a??? ?a? e?e??a p?? p???a????
  t?????t?ta. ?? p????f???e? a?t?? e??a?
  apa?a?t?te? ??a t? ??sp?s? ?a????? asfa????
  ???s?? t?? ??µ???? ??s??? ?a??? ?a? ??a t??
  e??µ???s? t?? ??????. ?p?????? d??f??a µa??µat???
  µ??t??a ??a t?? p??sd????sµ? t?? ep????d???t?ta?
  µ?a? ??µ???? ??s?a?.

63
?p?t?µ?s? ???d???? ap? t? ???s? ??????? ??s???

• ?p?????? as?fe?e? ?a? aßeßa??t?te? ??a t? ßa?µ?
  ep????d???t?ta? t?? ??µ???? ??s??? st?? ?????p?,
  af?? ?? d???µas?e? t?? in vivo t?????t?ta?
  ?????ta? se pe??aµat???a ?a? ta ap?te??sµata
  efa?µ????ta? st?? ?????p? ?at? p??s????s? ?a? µe
  ???e ep?f??a??. Se ???sµ??e? pe??pt?se??
  p??sd?????eta? ? ap?de?t?? ßa?µ?? ep????d???t?ta?
  µ?a? ??s?a? ? ?p???? ?a?????eta? ap? t? ????
  ??st???/?f??e?a? (??s?e? p?? s?µß?????? st??
  ??????µ??? a??pt???, ß??t??? ep?ped? ? p???t?ta
  ????).