PEPSIN

Pepsinum

Pepsin la che pham dang bot cha men tieu protein, lay t niem mac con ti cua da day ln, trau, bo hoac cu, tac dung trong moi trng acid (pH t 1 en 5), phai co hoat lc khong t hn 0,5 n v trong 1 mg, tnh theo che pham a lam kho.

ong vat lay bot pepsin phai ap ng cac yeu cau ve sc khoe ong vat phu hp s dung cho ngi va yeu cau cua c quan co tham quyen.

Phai ch ra phng phap a dung e loai bo s nhiem virus hay cac tac nhan gay nhiem khac.

Tnh chat

Bot ket tinh hoac vo nh hnh mau trang hay vang nhat, hut am. Tan trong nc thc te khong tan trong ethanol 96% .

Dung dch trong nc co the hi uc l va co phan ng acid nhe

nh tnh

Trong coi s cho 30 mg fibrin xanh (TT), tao hon dch vi 20 ml dung dch acid hydrocloric loang (TT). Loc hon dch va ra giay loc vi dung dch acid hydrocloric loang (TT) cho en khi dch loc khong mau. Choc thung giay loc va trang hon dch fibrin xanh qua lo thung vao mot bnh non bang 20 ml dung dch acid hydrocloric loang (TT). Lac hon hp nay trc khi dung. Hoa tan mot lng che pham khong t hn 20 n v hoat lc trong 2 ml dung dch acid hydrocloric loang (TT) va ieu chnh en pH 1,6 + 0,1 (dung dch th). Cho vao hai ong nghiem, moi ong 4 ml hon dch fibrin c ieu che tren. Them vao mot ong 1 ml dung dch th va ong con lai 1 ml nc (mau trang). Tron eu cac ong va cho vao noi cach thuy 25 oC, lac nhe trong 15 phut. Dung dch mau trang khong co mau va dung dch th phai co mau xanh.

Mat khoi lng do lam kho

Khong c qua 5,0% (Phu luc 9.6).

(0,500 g; phosphor pentoxyd; ap suat khong qua 0,7 kPa; 60 oC; 4 gi).

Gii han vi khuan

Tổng số vi khuẩn hiếu kh sống lại được: Khng được qu 104 khuẩn lạc/gam.

1,0 g che pham khong c co Escherichia coli va 10 g che pham khong c co Salmonella (Phu luc 13.6).

nh lng

Hoat lc cua pepsin c xac nh bang cach so sanh lng peptid, khong b tua bi dung dch acid tricloroacetic (TT) va c nh lng bang cach s dung thuoc th phosphomolibdotungstic, giai phong tng phut t c chat la dung dch hemoglobin, vi lng peptid giai phong t pepsin vi cung c chat va cung ieu kien.

Tranh lac va lam sui bot cac dung dch th va dung dch chuan trong qua trnh nh lng.

Dung dch th: Chuan b ngay trc khi dung mot dung dch cha 0,5 n v/ml trong dung dch acid hydrocloric loang (TT). Trc khi pha loang en the tch can thiet ieu chnh en pH 1,6 0,1 neu can bang dung dch acid hydrocloric 1 M (TT).

Dung dch oi chieu: Chuan b dung dch pepsin chuan (ĐC) cha 0,5 n v/ml trong dung dch acid hydrocloric loang (TT). Trc khi pha loang ieu chnh en pH 1,6 0,1 neu can bang dung dch acid hydrocloric 1 M (TT).

Ch pha trong vong 15 phut trc khi dung.

Chuan b cac cap ong nghiem T, Tb, S1, S1b, S2, S2b, S3. S3b va ong B.

Them dung dch acid hydrocloric loang (TT) vao cac ong nghiem nh sau:

B: 1,0 ml

S1 va S1b: 0,5 ml

S2 , S2b, T, va Tb: 0,25 ml

Them dung dch oi chieu vao cac ong nghiem nh sau:

S1 va S1b: 0,5 ml

S2 , va S2b, : 0,75 ml

S3. va S3b : 1,0 ml

Them 0,75 ml dung dch th vao ong nghiem T va Tb.

Them10,0 ml dung dch acid tricloroacetic (TT) vao cac ong S1b,S2b, S3b, Tb va B. Tron eu.

at cac ong nghiem va dung dch hemoglobin (TT) trong cach thuy nhiet o 25 0,1 C. Khi nhiet o a can bang them 5,0 ml dung dch hemoglobin (TT) vao ong nghiem B, S1b, S2b, S3b va Tb. Tron eu.

Them 5,0 ml dung dch hemoglobin (TT) lan lt va cach nhau 30 giay vao ong nghiem S1, S2, S3 va T. Tron eu ngay sau moi lan them.

ung 10 phut sau khi them dung dch hemoglobin (TT) dng phan ng bang cach them

them10,0 ml dung dch acid tricloroacetic (TT) vao ong S1, S2, S3 va T moi lan cach nhau 30 giay. Tron eu.

Loc cac ong nghiem (th va trang) qua giay loc thch hp a c ra trc bang dung dch acid tricloroacetic 5% (TT) sau o la nc va lam kho. Bo 5 ml dch loc au. Lay 3,0 ml moi dch loc rieng biet vao ong nghiem cha 20 ml nc. Tron eu.

(Giay loc thch hp la giay loc ap ng cac phep th sau:

Loc 5 ml dung dch acid tricloroacetic 5% qua mot giay loc trang, tron ng knh 7 cm. o o hap thu cua dch loc 275 nm (Phu luc 4.1), dung dung dch acid tricloroacetic 5% (TT) khong loc lam mau trang, phai t hn 0,04).

Toan bo qua trnh tren c ghi bang sau:

Ong nghiem

 

S1

S1b

S2

S2b

S3

S3b

T

Tb

B

Dung dch acid hydrocloric loang (TT) (ml)

0,5

0,5

0,25

0,25

 

 

0,25

0,25

1,0

Dung dch oi chieu (ml)

0,5

0,5

0,75

0,75

1,0

1,0

 

 

 

Dung dch th (ml)

 

 

 

 

 

 

0,75

0,75

 

Dung dch acid tricloroacetic (TT) (ml)

 

10,0

 

10,0

 

10,0

 

10,0

10,0

Tron

 

+

 

+

 

+

 

+

+

Cach thuy 25 oC

+

+

+

+

+

+

+

+

+

Dung dch hemoglobin (TT) (ml)

 

5,0

 

5,0

 

5,0

 

5,0

5,0

Tron

 

+

 

+

 

+

 

+

+

Dung dch hemoglobin (TT) (ml)

5,0

 

5,0

 

5,0

 

5,0

 

 

Tron

+

 

+

 

+

 

+

 

 

Cach thuy 25 oC 10 phut

+

+

+

+

+

+

+

+

+

Dung dch acid tricloroacetic 4% (TT) (ml)

10,0

 

10,0

 

10,0

 

10,0

 

 

Tron

+

 

+

 

+

 

+

 

 

Loc

+

+

+

+

+

+

+

+

+

Them vao moi ong nghiem 1,0 ml dung dch natri hydroxyd 20% (TT) va 1,0 ml thuoc th phosphomolybdotungstic (TT) bat au t cac ong trang sau la cac ong th theo th t nh tren.

Sau 15 phut o o hap thu (Phu luc 4.1) cua dung dch S1, S2, S3, S1b, S2b, S3b ,T va Tb bc song 540 nm dung dch loc cua ong B lam mau trang.

Hieu chnh o hap thu cua dch loc t ong S1, S2, S3 bang cach lay o hap thu o c tr i o hap thu dch loc S1b, S2b, S3b theo th t.

Ve ng cong chuan gia o hap thu a tnh va hoat lc dung dch oi chieu a dung.

Xac nh hoat lc cua che pham dung o hap thu cua dung dch th (T Tb) t ng cong chuan va tnh toan o pha loang.

Bao quan

ng trong lo kn, tranh anh sang va nhiet o t 2 en 8 C.