Dasar - Dasar

Toksikologi

Dr.Linda Dewanti, MKes, MHSc. PhD

IKM-KP FK Unair
References

 Richards IS. 2008. Principles and Practice of

Toxicology in Public Health. Jones and Bartlett
Publisher, Sudbury Massachusetts.

 Ernest Hodgson. 2004. A Textbook of Modern

Toxicology. Wiley-Interscience (3rd Edition)

 Principles of Toxicology: Environmental &

Industrial Application. 2000. William P.L. et al
 TOKSIKOLOGI

 Ilmu pengetahuan yang mempelajari efek negatif bahan

kimia terhadap sistim makhluk hidup
 Efek negative: segala bentuk perubahan yg menyimpang
dari “normal”
 Toksisitas (Toxicity):
Tingkat / Derajat suatu bahan kimia dalam
menimbulkan efek berbahaya bagi manusia atau binatang
* Akut
* Sub-kronik
* Kronik
 Toksikan (toxicant/poison): suatu bahan yang dapat
menimbulkan respon merugikan dalam sistem biologis
Apa saja Toxicant/racun?

2 beda pendapat
1) Semua bahan = toxicant
Dosis yg tepat bisa membedakan:
Toxicant & obat
Dosis: jumlah bahan kimia yg masuk ke tubuh

2) Toxicant: bahan yang dalam jumlah

sangat kecil sudah bisa menimbulkan
efek negatif dalam sistem biologis
Apa respons tubuh thd Toxicant?

Respons tergantung dosis & organisme /

individu
1. Berubah dari kondisi Normal
2. Lokal vs. sistemik
3. Reversible vs. irreversible
4. Immediate vs. delayed
Dose-Response Relationship:
Jika dosis response

RESPONSE

0-1 NOAEL
2-3 Linear Range
2 4 Maximum Response

0 1 DOSE
Dosis menentukan respon biologis
Mekanisme Pertahanan Tubuh thd
Toxicant

 Membran barrier: passive & facilitated

diffusion, active transport
 Biotransformation enzyme, antioksidan
 Mekanisme elimination
Toksisitas Akut:
Efek berbahaya bahan pada suatu organisme melalui
paparan tunggal / paparan singkat bahan kimia

Toksitas Sub-Kronik:
Kemampuan bahan toksik menimbulkan suatu efek
melalui paparan berulang dalam jangka waktu lama
tapi lama paparan < 0.5 harapan hidup makhluk
hidup

Toksisitas Kronik
Kemampuan suatu bahan (atau gabungan bbrp
bahan) menimbulkan efek bahaya dalam jangka
lama, pdu setelah paparan berulang / paparan terus
menerus (> 0.5 harapan hidup makhluk hidup)
 Ukuran Toksisitas

 Mortality (death): LD50, LC50

 Teratogenicity (ability to cause birth defects)
 Carcinogenicity (ability to cause cancer)
 Mutagenicity (ability to cause heritable
change in the DNA)
 Ukuran Toksisitas:
Median Lethal Dose (LD50 & LC50)

 LD50:
Jumlah / dosis bahan kimia yg
mengakibatkan kematian 50% populasi
hewan coba
=mg bahan kimia / kg berat badan hewan

LD50 Bahan A= 25 mg/kg

Toksisitas A >> B
LD50 Bahan B = 7.000 mg/kg
Ukuran Toksisitas LC50

LC50:
Konsentrasi bahan kimia di lingkungan
(udara/air) yang mengakibatkan kematian 50%
populasi hewan coba dalam waktu tertentu

mg bahan kimia / liter udara atau air (=ppm)

 Label Peringatan

CAUTION: LD50 >500 mg/kg

* > 5000 mg/kg = relatively non toksik
* 500 – 5000mg/kg = slightly toxic
WARNING: 50 mg/kg <LD50 < 500 mg/kg
(moderately toxic)
DANGER:
LD50 < 50 mg/kg (highly toxic)
POISSON
LD50 ≤ 50 mg/kg
 Relative Toksisitas Insektisida

 TCDD (Dioxin) 0.1 mg/kg

 Parathion 13.0 mg/kg
 Nicotine 50.0 mg/kg
 Carbaryl 270.0 mg.kg
 Malathion 370.0 mg/kg
 Relative Toksisitas
Herbisida & additive

mg/kg mg/kg
Paraquat 95 Limonene 5,000
2,4-D 375 Clopyralid >5,000
2,4-DP 532 Sulfometuron Met.. >5,000
Triclopyr 630 Imazapyr >5,000
Tebuthiuron 644 Diesel oil 7,380
Dicamba 757 Picloram 8,200
Hexazinone 1,690 Fosamine am.. 24,400
Glyphosate 4,320 Kerosene 28,000
 Toksisitas Relative Beberapa Bahan Kimia

Highly toxic chemicals Moderately toxic chemicals

0 – 50 mg/kg range 50- - 500 mg/kg range

mg/kg mg/kg
 Botulinus toxin 0.00001  Paraquat 95
 Dioxin 0.1  Caffeine 200
 Parathion 13.0  Carbaryl 270
 Strychnine 30.0  Malathion 370
 Nicotine 50.0  2,4-D 375
 Toksisitas Relative Beberapa Bahan Kimia
Slightly toxic chemicals: 500 – 5,000 mg/kg range

mg/kg
 Formaldehyde 800
 Hexazinone 1,690
mg/kg
 Asprin 1,700
 2,4-DP 532
 Vitamin B3 1,700
 Triclpoyr 630
 Household bleach 2,000
 Tebuthiuron 644
 Table salt 3,750
 Dicamba 757
 Glyphosate 4,320
etc.
Apakah semua bahan bersifat
relative toksik?

 Gula: LD50 = 30,000 mg/kg

 Ethanol: LD50 = 13,700 mg/kg
 Air: LD50 = 80,000 mg/kg
 Fase Toksikologi (3 fase):

 Paparan
 Toksiko-kinetik
* absorbsi
* distribusi
* metabolisme
* ekskresi
 Toksiko-dinamik
 Fase Toksikologi: PAPARAN

Bioavailability: fraksi dosis yang tersedia untuk

diabsorbsi

Faktor yg mempengaruhi:
*Waktu & frekwensi paparan
*Rute paparan (oral, inhalasi, kulit dll)
*Dosis
*Bentuk fisik & kimia bahan tsb (ukuran
partikel, kelarutan)
*Host
*Pre-absorpsion Metabolisme
 Fase Toksikologi: Absorption

 Aspek Komparatif: selluler, organisme

 Morfologi membran: lipoprotein bilayer
 Fisiokemikal proses dlm transfer antar
membran:
*Lipid-water solubility
*Ionisasi
*ukuran molekular
Fase Toksikologi: Absorption

 Trans-membran movement
*simple diffusion
*filtration
*carrier mediated
 Site of absorption (GI tract, dermal, paru)
 Fase Toksikologi: Distribusi

Bgmana distribusinya:
Darah, lympha
Barrier:
Blood brain barrier, placenta, mammary (milk)
Storage: adipose tissue (DDT) …..kelaparan: bahaya
tulang: Ca, Fl, Pb, strontium
Redistribution
Enterihepatic resirculation
 Fase Toksikologi: Metabolisme

 Bahan kimia yg masuk tubuh

dimodifikasi dg reaksi enzimatik:
* bahan kimia mjd lebih larut air mudah
di ekskresi
* lebih tdk larut lemak
Toksisitas
* meningkatkan ionisasi

 Conjugasi: berikatan dg soluble

endogenous agent
 Paparan secara bersamaan

Additive (2+2=4)
2 OP’s = cholinesterase inhibition
Synergistic (2+2=10): CCl4 + ethanol=
hepatotoxicity
Potentiation (2+0=6): isopropanol + CCl4 =
tumor promoter
Antagonism (2+2=0): BAL + Heavy metal
(Antidotum)
 Fase Toksikologi: Ekskresi

 Renal Excretion: urine

 Non Renal Excretion: empedu (feses),
exhalation (udara ekspirasi), susu, saliva,
keringat
Fase Toksikologi: Toxicodynamics

 Dose-response relationship
 Pengukuran Toksisitas

 Dilakukan di laboratorium
 Normal prosedur: memberi paparan pada
hewan coba
 Cara: memasukkan bahan kimia ke dalam
tubuh hewan coba (penelanan, inhalasi,
absorbsi kulit, sonde lambung, intraperitoneal,
injeksi intra-vena, subkutan, dll)

atau: memasukkan bahan kimia ke dalam air

minum atau udara
Toxicity Testing
Digunakan untuk:
* mengidentifikasi kemungkinan efek negatif paparan
thd suatu bahan
* membuat Dose Response Relationship (menjelaskan
SEVERITY efek paparan bahan yang sudah
diketahui)
* memprediksi efek paparan pada kelompok manusia
Konsensus desain Protokol Testing “meminimalkan” :
* Varians dan bias
* False + / False –
* Kesetimbangan antara informasi yg diinginkan dg
cost & resources nya
Macam Evaluasi Toksikologi:
1. Lama paparan: akut, sub-kronik,kronik
2. Target organ: general, spesifik
(reproduksi, imuno- , neuro-, genetik-,
carcinogenetik toksikologi)
 Materi / Bahasan

1. Hematotoxicity
2. Hepatotoxicity
3. Nephrotoxicity
 Serba serbi

Jaringan / organ Target

Konsentrasi toksikan tinggi di jaringan ≠

jaringan target
 Pb: >90% di tulang; ttp efek pada ginjal,
central & perifer saraf pusat,
hematopeietic system
Serba serbi

 Efek Neg. manifest pada spesifik organ

 CO (cardiac output) tinggi = paparan tinggi
 Tiap organ punya sel & jaringan khusus
 Proses di dalam sel & reseptor berbeda-beda
Protein- receptor match substrates
 Toksikan & metabolitnya mungkin punya reactive
pathway yg spesifik
 Toksikan tidak mengakibatkan efek Neg pada
semua organ dalam dosis yang sama
Serba serbi

 Sebuah toksikan mungkin punya

beberapa site of action & target organ
 Paparan multi-toksikan mungkin punya
terget organ yg sama (synergy)
 Target organ tsb mungkin bukan sebagai
storage
 Toksiko-kinetik menentukan konsentrasi
toksikan di target organ
PROSENTASE DISTRIBUSI ALIRAN DARAH
 Hematotoxicity:
Chemically Induced Toxicity of the Blood

Effects:
1. RBC (red blood cells)- mediated oxygen
Transport
2. the production of RBC, WBC. & platelets
 Efek thd RBC pdu terjadi melalui

1. Kompetitif inhibisi thd oksigen yang akan

berikatan dengan Hb (>>)
2. Anemia o/k bhn kimia (jumlah RBC yang
beredar menurun krn banyaknya RBC yg
rusak

Methemoglobinemia, a blood condition

characterized by cyanosis, was observed
when individuals consumed well water
containing large amounts of nitrates and
nitrates.
 Carbon monoxide, cyanide, and
hydrogen sulfide berikatan dg Hb & berpotensi
mengganggu kemampuan men-transport oksigen

 CO secara direct meng-inhibisi oksigen

utk binding ke Hb (y.i binding ke Fe 2+)
hipoksia (ringan s/d mengancam nyawa)
 Efek thd cardiovascular and neurological,
misal: delayed neurological injury:
memory loss, a reduction in
neuropsychological test performance).
CO

 CO akan melekat (binding) dg Hb

menurunkan ketersediaan tmp binding bg
oksigen; sementara itu juga meningkatkan
afinitas binding oksigen yg telah terikat pd
Hb (oksigen sulit di lepas dari Hb ke
jaringan yang membutuhkan) ≈ The Haldan
Effect
 Afinitas CO 200x (210-245x) lebih tinggi
dibanding O2
CO

 CO-Hb ≤ 1% Normal ada dlm darah

akibat metabolisme porphyrin
 Merokok (2 pak/hr) meningkatkan kadar
COHb mjd 5-10%
 Jk + paparan CO dari luar kadar COHb
mjd 20%
 Gejala Keracunan CO

 Hypoxia berat kronis brain injury dg Gx mirip

Parkinson’s disease, cognitive impairment, and serious
neurobehavioral changes.
 Keracunan ringan s/d sedang recovery akan pulih 100%
apalagi jk di Tx 100% oxygen atau hyperbaric oxygen
treatment
 Kadar COHb drh <10% (asimtomatik)
 Kadar COHb 15–20% mulai tjd hipoksia
 Kadar >20% tjd komplikasi cardiovascular and neurological
 Kadar >40%: headache, dizziness, nausea, & vomiting.
 Kadar >60% kemungkinan lethal
 Half Life CO

 Jk menghirup dalam ruangan, T1/2 dari CO

= ± 4–5 jam;
 Tx dg 100% oksigen, T1/2 turun 4x
 Tx hyperbaric oxygen, T1/2 turun 10x
Sumber pencemar CO

 Pembakaran tdk sempurna; gas buang kendaraan.

 Memakai pemanas rumah tanpa ventilasi yg tepat
 Paparan thd methylene chloride (cat pelapis
furniture). Methylene chloride akan dimetabolisme
oleh cytochrome P450 enzymes menjadi COHb
Aktivitas fisik Respiratory rate jml paparan
meningkat Physical activity, which increases the
respiratory rate, will increase the amount of inhaled
methylene chloride and the resulting
carboxyhemoglobin levels.
 TLV CO
 OSHA standard 50 ppm & ACGIH TWA 25
ppm
 50 ppm CO (setelah 8 jam) memproduksi
COHb 5-6% .

ANTIDOTUM
•Injeksi iv methylene blue (meningkatkan
kemampuan methemoglobin reductase)

Hyperbaric Oxygen Therapy

 Bahan kimia lain ≈ Meth-Hb:
INORGANIC NITRATES/NITRITES
& CHLORATE SALTS

 Inorganic nitrites spt: sodium nitrite (NaNO2) &

chlorates (ClO3−) mengoksidasi ferrous
hemoglobin (Fe 2+) mjd ferric-hemoglobin (Fe 3+
methemoglobin).
 Nitrate, direduksi mjd nitrite o/ bakteri nitrifikasi
dlm usus kmd merubah Ferrous mjd ferric
 Paparan di industri atau air minum
terkontaminasi nitrates, nitrites, and chlorates:
menimbulkan meth-Hb
Methemoglobinemia

 Kelainan yang disebabkan peningkatan

% methemoglobin (Hb + Fe 3+)
 Kemampuan untuk mengikat oksigen:
rendah
 Source: WashingtonEdu

Figure 1. Hemoglobin: Basic Structure

Hemoglobin is a protein contained within the red blood cells and it is comprised of
four globular protein subunits (heterotetramer). Each of these subunits contains a
heme group and the center of each heme group contains an iron, which is the site of
oxygen binding. This rendition shows hemoglobin A which consists of two alpha
and two beta subunits.
 figure: WashingtonEdu

Figure 2. Methemoglobin
Methemoglobin results from the change of the ferrous (Fe2+) state
to the ferric (Fe3+) state. The Fe3+ does not effectively bind oxygen.
METHEMOGLOBIN LEADING TO HEMOLYTIC ANEMIA:
AROMATIC AMINES & AROMATIC NITRO COMPOUNDS

 Aromatic amines and nitro compounds contoh aniline and

nitrobenzene menyebabkan methemoglobinemia by
initiating a redox cycle in the RBC & perubahan RBC
(kerusakan membran RBC) dikenali o/ splenic
macrophage dihancurkan
 Jika bone marrow tdk bisa mengkompensasi dg
meningkatkan pelepasan erythropoietin
hemolytic anemia
 Banyak digunakan di pengecatan, pharmaceutical
(dapsone, primaquine), & agricultural chemical industries.
 Dapsone and primaquine methemoglobinemia &
hemolytic anemia
Figure 4.4 Dapsone N-hydroxylamine-induced Red
Blood Cell Changes.
Chemically induced damage to red blood cells is typically
expressed as changes in red blood cell shape. The altered
shape (morphology) results from damage to the
cytoskeleton proteins or lipid membrane of the red blood
Pb & Anemia
 Pb anemia
Pb menghambat 2 enzim (ALA-
Dehydrogenase & ferrochelatase) dlm
heme synthesis pathway

 Anemia absorbsi Pb meningkat

(?)
 Iron Metabolism

Iron level

Signal to facilitate iron absorption

DMT1

Absorption of Iron & Pb

(Bannon et al., 2002)
By Andrews (1999)
 Chemicals Reported to Cause
Bone Marrow Suppression
 Benzene (an important
industrial solvent and
component of many refined
petroleum products, e.g.,
gasoline)
 Chloramphenicol (an important
antibiotic used to treat resistant
bacterial infections)
 Phenylbutazone
antinflammatory used to treat
arthritic conditions)
 Procainamide (an
antiarrhythmic used to control
cardiac arrhythmias)
 Allopurinol (a drug used to
treat gout)
 Tolbutamide (used to treat
 Methyldopa (an
antihypertensive used to treat
high blood pressure)
 Sulindac (antiinflammatory
agent)
 Aminopyrine (analgesic and
antipyretic)
 Sulfasalazine (a drug used to
treat inflammatory bowel
disease)
 Sodium valproate (used to
treat certain epileptic
conditions)
 Alkylating and antimetabolite
(cancer chemotherapy agents,
e.g., nitrogen mustard, 5-
fluorouracil, cytoxan)
 Chemicals Reported to Cause
Bone Marrow Suppression

 Isoniazid (a mainstay  Pentachlorophenol (a

antibiotic in treating chemical used to treat
tuberculosis) wood)
 Cephalothin (a  Carbamazepine (used to
cephalosporin antibiotic) treat certain forms of
 Gold (used as an epilepsy)
antiflammatory agent in
arthritic conditions)
 Diphenylhydantoin (an
important drug used in the
treatment of epilepsy)
HEPATOTOXICITY
 Fungsi Liver
■ Regulation of blood sugar. Kadar glukosa darah
dipertahankan 0.1%. Kelebihan akan disimpan dlm btk
glycogen
■ Biotransformation of chemicals. Merubah bahan yg tdk
larut lemak mjd bahan yang mudah larut air (lebih
mudah di ekskresi via urine)
■ Regulation of lipids. Lipid diambil dari darah dan
dirubah mjd karbohidrat atau yg lain ditimbun (fat
storage
■ Regulation of amino acids.
Kadar asam amino dlm darah dipertahankan dlm kadar
ttn. Asam amino esensial tdk bisa di storage shg di
rubah mjd bentuk urea dan di eskresi melalui urine
Hepatotoksikan

 Liver mempunyai kemampuan utk:

mengkonsentrasikan, biotransformasi &
ekskresi toksikan via empedu (potensial
tjd hepatotoksik)
 Pdu toksikan yg masuk melalui
penelanan (jarang di industri)
Target toksikan dalam sel hepar

1. Mitochondria (energy metabolism and

synthesis of ATP): carbon tetrachloride,
cocaine, dichloroethylene,
ethionine, hydrazine, and phosphorus.
2. Plasma membran (menjaga
keseimbangan ion dlm sitoplasma & di
luar sel): acetaminophen, ethanol,
mercurials, and phalloidin
Target toksikan dalam sel hepar

3. Endoplasmic Reticulum/ER (sintesis

proteins & phospholipids dlm hepatocyte).
Aktivitas Hepatic biotransformation enzyme
akan meningkat jk terpapar bhn kimia/obat,
tmsk cytochrome P450. ER = tmp
metabolisme oksidatif bhn kimia/ asing
(xenobiotic), shg jg mjd tmp dimana
metabolit yg reaktif terbentuk. Contoh:
acetaminophen, bromobenzene, carbon
tetrachloride, and cocaine
Target toksikan dalam sel hepar

4. Nucleus: bbrp toksikan/metabolit binding

ke DNA mutasi cell death, mjd
malignant/cancer. Ada toksikan:
mengaktivasi ensim endonucleases
(terletak dlm inti sel yg dpt men-digest
chromatin material uncontrolled
digestion of the cell’s DNA.
Contoh: aflatoxin B, beryllium, ethionine,
galactosamine, and nitrosamines.
Target toksikan dalam sel hepar

5. Lysosomes. Mengandung ensim digestive

(proteases) & penting dalam mendegradasi sel
mati / tua. Jk tjd injury pd hepatosit, lysosome
akan meningkat.

Tdk semua Paparan toksikan berakibat cell death,

apoptosis; bisa saja : morphologic abnormalities
cell (swelling, dilatasi endoplasmic reticulum,
condensed mitochondria & chromatin material
dlm nucleus. Bisa recovery o/k hepatocyte
punya kemampuan luar biasa utl regenerasi.
Target toksikan dalam sel hepar

 Ada bbrp yg irreversible injury: akibat

produk reactive metabolit nya
acetaminophen, bromobenzene, carbon
tetrachloride, chloroform, cocaine, and
trichloroethylene.
 Loss of Calcium Homeostasis
(Intracellular calcium sangat penting bg
semua proses dlm sel shg sangat diatur
ketat ratio calcium intracellular :
extracellular ≈ 1:10,000).
Target toksikan dalam sel hepar

 Calcium Homeostasis dilakukan o/

mitochondria and endoplasmic reticulum
 Jk keseimbangan tsb hilang disrupts
mitochondrial metabolism & ATP
synthesis, damages microfilaments yg
menopang struktur sel & aktivasi
degradative enzymes cell death
akibat hepatotoxic chemicals.
Immune Reactions of hepatotoxicity
 Kasus (jarang): halothane (gas anestesi)
dimetabolisme mjd reactive metabolite yg
berikatan dg protein. Protein tsb ter-
ekspresi pada permukaan sel & dikenali
sbg benda asing o/ sistem imun
destruction of the hepatocytes. Disebut
halothane hepatitis (50 % mortality rate). A
 Obat lain : diclofenac.
SUMBER:
Bbrp Bahan kima yg ditengarai /dicurigai “Potensial akan
meninmulkan efek macrovesicular steatosis yg mungkin
bisa mjd fatty liver”
 Cholestasis
(aliran empedu terhambat/terhenti )

 o/k hilangnya integrity dari sistem

canalicular (yg menyalurkan empedu ke
kandung empedu)
 α-naphthylisothiocyanate sambungan
antar rusak shg canaliculi (saluran terkecil
utk jalan empedu) tdk terbentuk sempurna
tumpah ke sinusoid sekitarnya
 o/k , while others are related to the
formation and secretion of bile
 Cholestasis
(aliran empedu terhambat/terhenti )

 methylene dianiline and paraquat,

merusak bile ducts.
 Anabolic steroids meng-inhibisi proses
transport bile acid dari darah ke dalam
lumen canaliculi
 Cirrhosis
 Chronic liver injury accumulation dari
collagen fibers dlm liver fibrosis. Akumulasi
Fibrotic tissue makin sukar bagi liver utk me-
replace sel rusak dg yg baru &
mempertahankan normal hepatic architecture.
Terbentuk dinding penyekat sel, micro-
sirkulatory terganggu hipoksia sel sel
mati terbentuk >> fibrotic scar tissue
cirrhosis aliran darah di liver obstruksi
portal hypertension shunts rupture
internal hemorrhage
 Tumor di Liver

 hepatocellular carcinomas
 Cholangiocarcinomas (berasal dr sel bile
duct)
 Hemangiosarcoma
 Adenomas
 cholangiofibromas
 Tumor di Liver

 Perubahan DNA mutations

 Nitrosoureas and nitrosamines
(hepatocarcinogens)
Masih dicurigai:
 anabolic steroids, arsenic, & thorium
dioxide
hepatocellular carcinoma
 Vinyl chloride, arsenic and thorium dioxide
Hemangiosarcoma