1) Carbohydrate

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EXOCRINE
1) Carbohydrate amylase 2) Lipid - lipase,
phospholipase A2, carboxyl esterase 3) Protein 
- trypsin, chymotrypsin, elastase
ENDOCRINE
(1) A(alpha or glucagon)??
(2) B(beta or insulin)??
(3) D(delta or somatostatin)??
(4) PP(pancreatic polypeptide)??
(5) G ?? (gastrin)
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Amylase? ??
1) a-Amylase ? Starch (??) ? Maltose ,
Glucose? ???? ? ?? , ?? , ??? ?? ?? ????
? ????? a-Amylase? ?? ??.
2) ß-Amylase ? Starch (??) ? Maltose ???
???? ? ??? ??
3) ?-Amylase (1) Starch (??) ? Glucose ???
????
Amylose, Amylopectin, Glycogen ?? ? ??? ???? ????
Dextrin? maltose, ? isomaltose, glucose ?
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Amylase? ??
1)?? ? ?? Amylase ?? ??? ???? ?? ?
?? Amylase ??? ??? ???? ?? ? ????? ???
???? ???? ??? ?? ??
2)??? ? Cl- , Br- , Ca2 , I2- , ... ... ...
? ???? ?? ? Cl- ?? ??? ? ???? ?? (0.01M
10mM ?? )
3)??? ? EDTA , Citrate , Oxalate , NaF ??
???? ? Ca2? ??? ?? ??? ????. ? Urea ,
Albumin, Lipemic Serum
4) Amylase ?? ?? Hormone Pancreozymin
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Amylase ????
a-amylase a-1.4 - linkage ? ??? hydrolysis
ß-amylase a-1. 4 - linkage ? ??? ??, maltose???
??
a-glycosidase a-1.4-linkage? ?????, glucose???
??
Isoamylase a-1.6-linkage? ????
Cellulase 1.4-ß-glycoside linkage ????
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???? ??

• 19?? ? ? ????(?? ????)

• 1831? ? Leuchs ???? ??(???)

• 1845? ? Bouchardat ?? ?? ??(????)

• 1895? ? Beyerrinck Amylase (??????)

• 1908? ? Wohlgmuth Iodometric method(B,U)

1925? Kuhn? a-amylase? ß-amylase??? ??

• 1916? ? Stocks ????? ??

• 1929? ? Elman Acute pancreatitis(early stage)

• 1934? ? Katsch ????

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• Amyloclastic method (Iodometric)

???? ??? Iodometric???? ?-  albumin ? ???
?? ??? ??. Wohlgemuth M, Van-Loon M,
Caraway M,

• Saccharogenic method

??? ??? ??? ???? ??- ??, ??, ?? Somogyi M, Meyer
M, Henry M, Sax M, Anthron M, dinitrosalicylic M

• Chromogenic method

??? ??? ???? ?? ??? ??  ??? ??? ??. ???? ??,
Blue starch?, Dyamyl ?, Amylochrome?

• Oligoglucose method

????? ???? ??? ?? ???? ?? ???, ? ????? ??

• ? ? ? ? ?

4-nitrophenyl, G7-4-NP, G5-4NP ? ??????? ?? ??,
4-NP? ??? ??? a - amylase? a-glucosidase???????
 PNP? ????
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• Amyloclastic method (Iodometric)

Caraway method
10mg / 100ml /37? / 30min
 Winslow method    
Wohlgemuth
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• Saccharogenic method

Somogyi-Nelson
1mg/100ml/37.5C/30min
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• Chromogenic method

Blue starch method
Cibachron blue F3GA
blue starch polymer
???? ??? ???? ???? ?? ??
DyAmyl-L Method
Amylopectin ???? ??
Reactone red 2 B
ZnCl2 -ethylene glycol monomethyl ethter
 ?? ?? ?? ???? ?? ???? ??
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• Oligoglucose method

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• ? ? ? ? ?

P-nitrophenyl maltohepatoside( p-NP-G7)
Maltopentaose oxidase
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?? ?? ? ??
CARAWAY- METHOD
??? ??? ?? ???? ?? (? )? ???? ?? ? amylase? ??
??? ????. ?? ?? ?? ???? ?? ??? ???? ????(???)
????.
Amylase Starch Substrate(pH 6.8-7.2)
Na2HPO4       26.6g Benzoic Acid     8.6g
. Starch 0.4gm. ??? 500ml
1/10N Stock Iodine solution
 Pot, iodine(KIO3) -1.785g,     Pot, iodide(KI) -
22.5g H2SO4 - 4.5 ml ??? - 500 ml
 1/100N Working Iodine solution
 Stock iodine sol(1/10N) 50ml? ??? 500ml?? ??
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PROCEDURE
TEST BLANK
Substrate 1.0? 1.0?
Pre warming 37? //5min Pre warming 37? //5min Pre warming 37? //5min
Serum 0.02 ? 0.02?(DW)
??? 7? 30? incubation ??? 7? 30? incubation ??? 7? 30? incubation
?? ?? 1.0? 1.0?
? ? ? 5.0? 5.0?
660? against reagent blank set to zero absorbance 660? against reagent blank set to zero absorbance 660? against reagent blank set to zero absorbance
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Normal Value
60 180 Somorgy unit/100 ?
70 2500 somorgy
Unit /1hr urine
1200 - 5000 Somogyi Unit / 24hr urine     50
170 Caraway Unit/100 ?         

• Somogy ?? ?? 100??? ?? 37 ?, 30? ? ??
• ? ???? ??? ????
• ????(IU)  Somogy? ?? ??? ??

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??? ??
1)?? Amylase (S? Amylase)
? ? ? ?
??? ?? ????, ?? ?, ??? ???? ??? ??? ?? ?
2)?? Amylase (P? Amylase)
? ? ? ?
????? ??, ???? ?? ??? ????? ???? ?? ???, ???
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????
Citrate, Oxalate, EDTA, NaF,- ?? ??(Ca??)
Bilirubin, hemoglobin ???? ??. Triglyceride?
500mg/dl ??? ??? ?? ??.
?????, Aminosalicylic  acid, Asparaginase,
Azathioprine, Corticosteroid,
Cycloheptadine, Nacortic analgesis, 
Rifampin, ???, ???. Sulfasalazine,
flouride ?? ??.
????? ?? ? 4-12??? ?? ?, 48-72?? ??  ???? ??.
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Iso-Electrophoresis
Iso-enzyme AMS
Subunit S P
Isoenzyme S1 - S3, P1 - P3
Buffer Veronal buffer pH8.6
??? CA
Stain Blue starch agar 2 Blue starch 1tab/5ml
Volume 1 ? /cm
???? 350V/2-3hr/5C
nm 600?
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LIPASE
? ? ? ? ? ? ? ?
? ? ? ???? ??? ? 1,000 / 0.2?/min (38?) (400?) 15Unite ?
???? ? 0.05N NaOH ??? (?) / 1? / 24hr 0.10.6/24hr 0 1 /8hr 0.334 0.16/3hr 0.32 0.13/2hr
?? ?? ? ??? free Fatty acid (?ol) /1? /1hr ???? free fatty acid ?ol /l /min 1.1 0.4 unite 60 76 unite
?? ?? ? p-naphthyl laurate, a-naphthyl palmitate. ???? ??? ?? ????? ??? ??? ????? ?? ???  esterase? ??? ???.
???? ?? anti human ab?sandwitch method(peroxidase)
? ? ? Diglyceride, BALB, U/L 1,2-O-dilauryl-rac-glycerol-3-glutarate
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Lipase Substrate
1,2-O-dilauryl-rac-glycerol-3-glutaric acid
(6-methylresorufin) ester
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???? ?
 Cherry-Crandall
 Tietz method
?? 1??? ??? ??? ???? ?? ?? Fatty acid? ????? ???
0.05N NaOH? ??
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 Tietz method
 ???              75 ml           Sod
benzoate      0.15gm          Arabia Gum        
5.25gm      Olive Oil           75 ml 
  tris(hydroxylmethyl)aminomethan.....48.55gm
  Distil Water    ...............................
..500ml
Stock sol..............................50ml 1N
HCL...............................21.5ml  (Ajust
pH 8.0)   Distill water.........................20
0ml
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Unknown Blank
??? 3.0? 3.0?
Olive oil 2.0? 2.0?
Tris-buffer 0.5? 0.5?
Serum 1.0? -
? ???? 6hr/37? ? ???? 6hr/37? ? ???? 6hr/37?
Serum 1.0?
Thymolphthaline 3drop 3.drop
Methanol 3.0? 3.0?
 ? ???, Blank ??? ?? 0.05N NaoH? ??(Blue)
?   Unknown??? ?? ???? ???? NaOH? ???? ????.
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Lipase -Enzymatic method - 1
R1 BICIN buffer 50mmol/L, pH 8.0
(250?). (2.0?) Colipase(porcine
pancreas) 0.9mg/L
Na-deoxucholate 1.6mmol/L
calcium chloride 10mmol/L
detergent preservative
R2 Tartrate buffer 10mmol/L, pH 4.0
(200?) 1,2-O-dilauryl-rac-gl
ycerol-3-glutaric acid (6-methylre
sorufin) ester 0.27 mmol/L
Taurodeoxycholate 8.8mmol/L
detergant preservative
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LIPASE - Enzymatic method - 2
Vogel-Zieve
Shihabi-Bishop
The decrease in turbidity is measured at 340nm
1) Buffer - Tris buffer 26mmol/L, pH 8.9 2)
Substrate sod deoxycholate
16.7mmol/L calcium chloride
0.04mmol/L Triolein0.3mmol/L
colipase 4mg/L 3) Standard - lipase
activity stated on vial in U/L/37C
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Procedure
Standard Test Control
Reagent 2.5 ? 2.5 ? 2.5 ?
Standard 0.1 ?
Sample 0.1 ?
Control 0.1 ?
Mix Avoid the formation of foam. Read absorbance
A1 of standard, control and sample after 4min.
After a further 5min read absorbance A2 of
standard, control and sample. 340nm
?A of standard, control and sample. A1 A2
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LIPASE - Enzymatic method - 3
????? 1.2-Diglyceride? ???? lipase? ?? ????
2-Monoglyceride? Quinone color ??
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Reagent-1
1.2-Diglyceride, Monoglyceride lipase(MGLP) Glycer
ol kinase(GK) Glycerol-3-phosphate
oxidase(GPO) Adenosine-3-phosphate(ATP) N-ethyl-N-
(2-hydroxy-3-sulfoprophyl-m-toludine(TOOS)
Reagent-2
4-Aminoantiphyrine Peroxidase
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PROCEDUER

1. ?? 5?? ?? ??? 300? ??, ? ?? ? 37??? 5min ?
   incubation

2) 5? ? ??? 100?? ??? ?? ?, 3-5? ??? ???
546? ?? ??? ??? ?? ??
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? ?
???? 1.2-diglycerid?  ???? ?? ????? ?? ?? ?
?? ?Colipase ?  Deoxycholine ? ???? ????? ??
lipase ? ???? ??? Esterase  Liver
lipase, lipoprotein lipase ? ???
??   ?Free glyceride, ascorbic acid ? ??? ???
???? ?? ??? ???  ??