Nhdn xel • Kit qua khao sat cho thiy phuong phap cd dp dimg cao (ty le tim lai tir 99,48%-100,85%) dam bao tot cho viec dinh luong levodropropizin trong che pham.
3 2.6. Khoang xde dinh
Trong phln danh gia dp dung, dp myin tinh, d cac miic ndng dp 80% va 120% ham Iupng boat chit tim lai dam bao kha nang tim lai, dp chinh xac va dp hiyin tinh. Vay khoang xac dinh cua boat chat la tir 80% din 120% so vdi lupng ghi tren nhan. Kit qua dupe trinh bay d phln danh gia dp dung 3.2.5, danh gia dp tuyin tinh 3.2.3 va danh gia dp ehinh xac 3.2.4.
TM li^u 4. Ket lu|ii
Qua khao sat thuc nghiem, chiing toi da xay dung dupe mdt chuang trinh sac ky long hilu n5ng cao di dinh lupng levodropropizin trong thudc vien nen.
Phuang phap xay dung cd su tuang quan tuyen tinh chat che giiia dien tich pic dap ung v^ ndng dO hoal chit, dp lap lai, dp chinh xac trung gian va dO diing cao vdi cac dieu kiln trang thilt bi, hda chat, thufic thir phu hpp vdi da sd cac phdng thi nghiim, Vai cac kit qua thu dupc, chung tdi di nghi cd the ap dung phuang phap dinh luong da nghiln cuu cho ch6 phlm thudc vien nen cd thanh phan levodropropizin.
^u tham khao
1. Vien Kiim nghiem thuoc Trung ucmg - Bo Y ti (2010), Ddm bdo chdt lirang thuoc vd mot so phuang phdp kiem nghiem thudc (Tai lieu dao tao nang cao vi kiim nghiem thudc), Nha xult ban Y hoc, trang 158-179.
2.DuocdiinAnh2013.trm%\Z2\-n22.
3. Dirac diin Hdn Qudc IX (2007), trang 561-563.
4 r/jeMefcft/nrfej:. thirteenth edition (200!), p 609.
5. http://www.benhhoc.com/thuoc/1405-Levopront.html.
SUMMARY
A specific HPLC method was developed and proposed for identification and quantitation of levodropropizine in Leprozine tablets.
Chromatographic conditions are as follow:
- Mobile phase: Solution A - Methanol (75:25)
(Solution A: Potassium phosphate monobasic 0.05Min water. Adjust to pH 3.0 ±0.1 with 85% phosphoric acid).
- Detector UV: 254 nm
- Column Phenomenex Rp 18 (250x 4.6 mm; 5 fim) - Flow rate: LO ml/min
- Injection volume: 20 /JI
The method was validated about the system suitability, specificity, linear range, precision, accuracy, and the validation results proved that this method was suitable for identification and quantitation of Levodropropizine in Leprozine tablets.
NGHigN CCrU DjNH Ll/ONG PROMETHAZIN HYDROCLORID TRONG SIRO PROMETHAZIN BANG PHI/0NG PHAP HPLC
TRINH THI HONG THAO, CAO NGOC ANH, TRAN THI BICH VAN Vien Kiem nghiem thuoc Trung uffng
I. Dat van de
Promethazin hydroelorid thudc nhdm thude khang Histamin (thu the HI), Promethazin dupe diing di chira trieu chiing hoac de phdng eae phan img qua min (nhu may day, phii maeh, viera raiii, viera kit mac va ngiia), de an than d ca tre em (trir tre nhd) va ngudi ldn. Hien nay tren thi trutrng da xult hien nhieu chl phara chiia Proraethazin hydroelorid. Trong
Dupc diin My 34 [3] hay Dupc diin Anh (BP. 2010) [1], Promethazin hydroelorid dupc dinh lupng bang phuang phap chiit do quang UV-VIS; Dupc dien Chau Au EP 7.0 [2] Proraethazin hydroelorid diiQC djnh lupng bang phuang phap chuin dp. Phuang pbSp chuan dp va phuong phap chilt do quang UV-VIS cd the gap mpt sd khd khan: nhu mit mlu trong qui trinh chiit, thao tac chiit phirc tap, kit qua dinh luong Tap chi KIEM NGHIEM THUOC - S6 4.2014; TSp 12.(4*)
phy thudc nhieu vao kiim nghiim vien, tdn nhieu thdi gian tien hanh phan tich. Nhung yeu td nay deu cd su anh hudng den kit qua dinh luong. Vl vay de dap ling nhu eau kiem nghiem cac che pham cd chiia Promethazin hydroelorid, chung tdi xin gidi thieu ket qua nghien ciiu dinh lupng Promethazin hydroelorid trong sird PROMETHAZIN bang phuong phap sac ky long hieu nang cao.
2. Thirc nghiem
2.1. Thiet bi, hda chdt, chdi chudn 2 Ll. Thietbi, dungcu
Da dupc hieu ehuin theo ISO/IEC 17025.
- May HPLC Agilent 1200 vdi detector DAD;
- May HPLC Agilent 1260 vdi detector DAD;
- C§n phan tieh Mettler Toledo AB 204 cd dp chinh xac 0,1 mg;
- Can phan tich Mettler AE 163 cd dp chinh xac 0,01 mg;
- Cac dung cu thuy tinh chinh xac: binh dinh muc, pipet.
2.1.2. Chdt chudn
- Promethazin hydroelorid: Chat chuan cua Vien Kiem nghiem thudc Trung uang; SKS: 0204085;
Ham lupng: 99,65% (khan), dp Ira: 0,13%.
2.1.3. Hoa chdt, thuoc thir: dgt tinh khiet phdn tich, linh khiet sdc ky
- Triethylamin (PA; Merck);
- Methanol (HPLC, Merck);
- Acid acetic bang (HPLC, Prolabo);
- Acid hydrocloric (PA, Merck).
2.2. Ddi tirong vd phirong phdp nghien ciru 2.2.1. Doi tircmg nghien ciru
- Mlu thir: Sird PROMETHAZIN: Cdng thuc ed trong 100 ml che pham: 100,0 mg Proraethazin hydroelorid. Ld SX: 030913, NSX: 090913, HD: 090916. Nha san xult: Cdng ty CP Dupc TW Vidipha.
- Mau gia dupc (placebo): Cdng ty CP Dupe TW Vidipha cung elp
2.2.2. Phircmg phdp nghien ciru 2.2.2.1. Dieu kien sic ky
- Cpt: Rpl8, 5 pm (250 x 4,6 mm) - Detector: 254 nra
- Tdc dp ddng: 1,5 ral/rain
- Pha ddng: Methanol - Dung dich dim pH 2,3 (60:40). Dieu chinh ty le neu can.
Dung dich dera pH 2,3: Them 2 ml triethylamin vao 1000 ml nudc, dilu chinh pH den 2,3 bang acid acetic bang.
- Thl tich tiem: 20 i^l 2.2.2.2. Phuong phap xir Iy mau
Dung djch chuan: Can chinh xac 20,0 mg Promethazin hydroelorid chuan vao binh dinh miic 100 ml, hda tan bang acid hydrocloric 0,1M them vita du din vach vdi ciing dung mdi, trpn deu. Hut chinh xac 5,0 ml dung dich nay, pha loang vira du 50 ml bang acid hydrocloric 0,1M, lpc qua raang lpc 0,45 |im, thu dupe dung dieh ehuan cd ndng dp 0,0199 rag/ml.
- Dung dieh thir: Can chinh xae mpt lupng sird tucmg umg vdi 10 mg Promethazin hydroelorid vao binh dinh mirc 50 ral, pha loang bang dung dieh aeid hydrocloric 0,1M vira dii den vaeh, trdn deu. Hiit ehinh xae 5,0 ml dung dieh tren vao binh dinh miie 50 ml va pha loang bang dung djch acid hydrocloric 0,1M vira du den vach, trdn deu. Lpc qua raang lpc 0,45 |im.
- Dung dich placebo: Tien hanh nhu dung djch thir nhung thay raau thii bang lupng raau placebo tuong duong vdi lupng che pham cd chiia 10 mg Proraethazin hydroelorid.
3. Ket qua va ban lu^n 3.1. Chgn diiu kien sdc ky
Chiing tdi da khao sat (thay ddi cdt, tdc dp ddng, pha ddng, ty le pha ddng) va lua chpn dupe dieu kien sac ky nhu da neu d phan 2.2.2.1, sac ky do thu dupc ciia dung dich ehuan va thu the hien d Hinh 3 va Hinh 4.
Tren sac ky dd chiing ta thay, pic Promethazin hydroelorid dupc tach hoan toan va can ddi, thdi gian luu khoang 4,3 phiit.
3.2. Thdm dinh phwang phdp 3.2.1. Tinh ddc hieu
Tiem vao he thdng sic ky lln lupt cac dung dich sau: Dung djch mlu trlng (dung dich acid hydrocloric 0,1M), dung dich mlu placebo; dung dich ehuan va dung dich thu. Kit qua khao sat tinh dac hilu dupc thi hien d Hinh 1, Hinh 2, Hinh 3, Hinh 4 va Hinh 5.
Tap chi K I £ M NGHIf M THUOC - So 4.2014; T3p 12.(46)
Hinh 2. Sdc ky do placebo
1 1
1 — V- • s • • —I ; • • • ; ,
Hlnh 3. Sdc ky do dung dich chudn tj^ Promethazin hydroelorid -4,3phul
Hlnh 4. Sdc ky do dung dich thu tj^ Promethazin hydroelorid ~4.2phut
— *.IWA:==-^^ - - Tren sic ky dd dung dieh mlu trang va dung dich mlu placebo khdng xult hien pic d trong khoang ,': thdi gian luu tucmg iing vdi thdi gian luu ciia pic
Promethazin hydroelorid.
Nhu vay phuong phap sac ky dinh lupng Promethazin hydroelorid vdi cac diiu kiln da xay dung dac hieu.
Hinh 5. Phd UV cua pic Promethazin hydroelorid trong 3.2.2. Tinh thich hap ciia he thong sdc ky dung dich thir vd dung dich chudn
Kit qua eho thay:
- Tren sic ky dd dung djeh thu cho pic cd thai gian luu va phd UV gidng thdi gian luu va phd UV cua pic Promethazin hydroelorid tren sic ky dd cua dung dich chuan (khoang 4,3 phut).
Tiem lap lai 6 lan dung dich ehuan Promethazin hydroelorid cd ndng dp la 0,02 mg/ml. Ghi lai cac %\i trj ve thdi gian lim va dien tich pic, sd dia ly thuylt vi he sd ddi ximg cua cac pie tucmg ling. Kit qua dupc thi hlln d Bang 1.
Bdng 1. Kel qud khdo sdt tinh thich hap ciia he thong sdc ky
TT 1 2 3 4 5 6 Trung binh
RSD (%)
Thai gian luv (Phiit) 4,268 4.270 4,269 4,272 4,272 4,266 4,270 0,05% (< 1,0%)
Dien tich pic (mAU.s) 1826,9 1825,0 1803,7 1826,1 1822,6 1822,1 1821,1 0,48% (< 2,0%)
S3 dta ly thuyit (N) 7005 7055 7134 6928 6961 6936 7003
//f so doi xung (T) 1,18 1,16 1,18 1,15 1,15 1,18 1,17
Kit qua khao sat eho thay, RSD cua thdi gian iuu nhd han \% va dien tich pic nhd han 2%», chimg td he thdng sic ky trin thich hpp de dinh lupng Promethazin hydroelorid trong sird PROMETHAZIN.
Tap chi KIEM NGHIEM THUOC - So 4.2014; TSp 12.(46)
CHAT CHUAN DOI CHIEU U S f T^l VIET NAM USP Reference
Standards
USP Dietary Supplement ^^m^^^m 1! USP Food Ingredient
Standards ^ ^ O ^ ^ ^ E . ^ ^ Standards
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Pharmacopeial Forum (PF)
USP P F
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I FOOD FRAUD DATABASE
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Tham J^guyen Traiting Ci.
34/6 Ydn Thtf, Q. TSn Binh, Tp. HCM, Vl$t Nam Tel: (84-6) 3848 8246 (Slines) * Fax: (84-8) 3848 5653
Website: www.phamnguyen.com.vn Email: [email protected] „
Sl
3.2.3. Khodng tuyen tinh
Chuin bi 5 dung dich chuin, cd ndng dp 50%; 80%; 100%, 120% va 150% so vdi ndng dp dinh luong. Tiin hanh sac ky cac dung dich chuin, ghi lai cac sac ky do va xac dinh dap iing ciia pic. Ket qua dupc ghi lai trong Bang 2 va Hinh 6.
Bdng 2. Ket qud khdo sdt khodng tuyen linh STT
1 2 3 4 5
Miu
50%
80%
100%
120%
150%
f^ong dd dung dfch chuan gac (mg/mi) 0,1990
DSpha loang
50/2,5 50/4,0 50/5,0 50/6,0 20/3,0
Ndng dp (Mg/mi
9,9520 15,9233 19,9041 23,8849 29,8561
Difn tich pic (mAU.s)
910,5 1439,4 1821,1 2185,4 2710,5 Phmmg trinh boi quy: y = 90,8841x + 4,4151
He s6 tirong quaa: R = 0,9999
S
'""•^'I'SX'."
'"-^
^ ^ -^
Nong do lM8/ml)
Hinh 6. Do thi bieu dien moi tuang quan giira nong do (pg/ml) vd dien tich pic ciia Promethazin hydroelorid Kit qua cho thSy vdi dieu kien sSc ky dS lira chpn, trong khoang ndng dp da khao sat tir 9,9520 |ig/ml den 29,8561 pg/ml, cd sy tuong quan tuyen tinh chat che giira ndng dp cua Promethazin hydroelorid va dien tich pic dap uHQg vdi be sd tuang quan la 0,9999.
3.2.4. Do chinh xac ciia phuang phdp
- Dp lap lai: Dp lap lai cua phuang phap duoc danh gia dua tren dp lech chuan tuong ddi cua 6 mau thu dupe phSn tich rieng biet theo qui trinh phan tich d muc 2.2.2.
- Dp chinh xac trung gian: Dp chinh xac trung gian dupc danh gia dua tren dp lech chuan tuang ddi cua 12 mlu thli dupc phan tich rieng biet tren hai thiet bi d 2 ngay khac nhau va hai KNV khac nhau. Ket qua khao sat dp l^p lai va dp chinh xac trung gian duoc the hien d Bang 3.
Bang 3. Kit qud khdo sdt do chinh xde ciia phuong phdp
TT
1 2 3 4 5 6
KNVl Lufmg cdn chudn: 20,0 mg Ti tr^ng mdu thti": l,29g/mi Thiit bi: Mdy HPLC Agilent 1200 Dim tich pic TB mdu chudn: 1821,1 (mA U.sj
LupKg c3n thif (g) 12,8941 12,9001 12,9130 12,9338 12,9476 12,9565
Difn tich pic (mAU.s)
1844,7 1855,5 1858,0 1869,3 1866,2 1858,5
Ham iuffng
(%)
100,9 101,4 101,4 101,9 101,6 101,1 Trung binll = 101,4%; n = 6; RSD - 0,36%
KNV2 Lircmg cdn cliudn. 19,8 mg Ti trgng mdu thir- 1.29g/ml Thiit bi: Mdy HPLC Agilent 1260 Difn lich pic TB mdu chudn-1739,4 (mAU.s)
LiepTig can thif
12,8931 12,9015 12,9064 12,9129 12,9197 12,9203
Dien tich pic (mAU.s)
1820,7 1806,0 1804,1 1812,8 1814,1 1798,1
Hilm iuf/ng
(%)
103,2 102,3 102,1 102,6 102,6 101,7 Trung binh = 102,4%; n - 6; RSD - 0,49%
Ket qua dinh lirgng tmng binh 12 mlu thir = 101,9%; n = 12; RSD = 0,67%
Nhu vay phuong phdp dat yeu cau vi dp chinh xac, dp lap lai trong ngay va khac ngay cua Promethazin hydroelorid la 0,49 % v^ 0,67 %.
3.2.5. Dodung
Dp dung cua phuang phap dupc tien hanh theo phuang phap them chuan vao mau placebo.
T?p chi KIEM NGHIEM THUOC - So 4.2014; Tkp 12.(46)
Can chinh xac khoang 8 mg, 10 mg, 12 mg Promethazin hydroelorid chuan vao binh dinh muc 50 ml da chiia sin mdt lupng gia dupc (khoang 12,9 g gia dupc), hda tan hoan toan bang acid hydrocloric 0,1M, pha loang bang dung dich acid hydrocloric 0,1M vira du den vach, trdn deu. Hiit chinh xac 5 ml dung dich nay pha loang vira du
bang 50 ml acid hydrocloric 0,1M, lpc qua mang lpc 0,45 fim, thu dupc cac dung dich cd nong dd khoang 80, 100 va 120% so vdi nong dp dinh lupng.
Moi miic ndng dd lam 3 mau.
Dien tich pic trung binh ciia Promethazin hydroelorid trong dung dich chuan la 1821,1 mAU.s
Kit qua khao sat dp diing duoc the hien d Bang 4.
? 4. Kit qud khdo sdt do dung cua phuang Mau
80%
100%
120%
Lumtg cdn placebo (g)
12,8133 12,9105 12,9318 12,9845 13,0263 13,0364 13,1368 13,1877 13,3679
Ltetmg Promethazin hydroelorid chuan
them vao (mg) 8,22 8,33 8,39 9,98 10,09 10,12 12,05 12,11 12,22
Dien tich pic (mAU.s) 1510,5 1520,4 1531,8 1822,6 1827,2 1833,3 2208,4 2212,8 2223,7
Luffng Promethazin hydroelorid ttm lai
(mg) 8,29 8,35 8,41 10,01 10,03 10,07 12,13 12,15 12,21
%Hdi phuc
100,9 100,2 100,3 100,3 99,4 99,5 100,6 100,3 99,9
Trung binh
TB = 100,5%
RSD = 0,38%
TB = 99,7%
RSD = 0,48%
TB -100,3%
RSD-0,36%
Tt-ung binh = 100,3%; n = 9; RSD = 0,48%
Nhu vay phuong phap phan tich dat yeu cau ve dp diing, ty le thu hdi ciia Promethazin hydroelorid tu 99,4% din 100,9%.
4. K€t luan
Tu ket qua khao sat tren da xay dung dupc phuang phap sac ky ldng pha dao de dinh lupng Promethazin hydroelorid trong sird PROMETHAZIN. Phuang phap su dung cdt Rp 18 ^250 x 4,6 mm; 5 )j.ra); Tdc dd ddng 1,5 ml/phiit; Th8 tich tiem 20 |il; Pha dpng
la hdn hpp dung dich dem pH 2,3 - Methanol (40:60);
Detector 254 nm. Phuang phap da dupc tham dinh vi tinh dac hieu, tinh thich hpp cua he thdng sSc ky, tinh tuyen tinh, dp diing, dp lap lai, dp chinh xac tnmg gian. Ket qua tham dinh cho thay phuong phap HPLC da xay dung hoan toan phii hpp cho viec dinh lupng Promethazin hydroclond trong sird PROMETHAZIN va cd the ap dung dinh lupng boat chat nay trong cac che pham thudc tuong tu.
l.S;^20;0. pp. 3038-3041.
2.£/'7.0(2011),pp.2804.
3. USP. 34 (2011), pp. 4039 - 4040.
Tai ligu tham khao
SUMMARY
Reversed'phase liquid chromatography method was developed for assay of Promethazine hydrochloride in PROMETHAZINE syrup. The method was earned by using a column Rp!8 (250 x 4.6 mm; 5 fim); the flow rate of mobile phase is maintained at I 5 ml per minute, injection volume: 20 pi. A mature ofpH 2 3 buffer solution and Methanol (40:60) as mobile phase. Detector set up at 254 nm. The method was validated in specificity, linear range; accuracy, precision. Validation results proved that the developed method was suitable for assay of Promethazine hydrochloride in PROMETHAZINE syrup
T a p chi K I £ M N G H I E M T H U O C - So 4.2014; T^p 12.(46)
