Khao sat de on dinh cua rhein trong huyet twang a nOng dO khoang 0,3 va 7,9 pg/mL trong dieu kien: bao quan a nhiet de ph6ng trong 6 gia; trong 24 gib, trong bum rn'eu to Ong a 15°C, sau 3 chu kjt doling - re Ong; bac, quail a nhiet de -20 °C trong 30, 60, 90 ngaỵ Mau duvc xem IA On dinh khi kat qua dinh luvng cac m5u sau thai gian bao quan dpt tir 85 - 115 % so vai gia tri ban deụ
Ket qua tem tat trinh bay trong bang 4 cho they phirang phap phi) hap vai y6u ceu de On dinh cua nghien cirụ
Bang 4: Kat qua khao sat ă) on dinh cua rhein trong huyet tut:mg
Digo kiOn dO 6n dinh DO loch (%) so voi ban dáu 0,29 pg/mL 7,87 pg/mL DO On dinh ngOn hpn
Sau 6 gib b nhiOt dO ph6ng 5,99 6,81 Sau 24 gi6 6' nhiPt d6 15°C
trong barn inPu ty, dOng 3,30 8,53 DO On dinh dong-r5 (long 8,67 5,92 DO On dinh dai han
CY -20°C sau 15 ngay 11,18 8,62
(a_11 -20°C sau 35 ngay 0,87 8,72
CY -20°C sau 60 ngay 0,08 4,21
Ban Wan
Phtyang phap HPLC dOng dinh luvng rhein trong huyet tirang ngkrai duvc xay dkrng vai de nhpy va de chinh xac caọ Theo Wei Zhu va Xue- Mei Wang [4] de sir dung phuceng phap chiet long-
long bang h6n hap diethylether-cyclohexan tjt le 4:1, trong nghien cÚu chi:mg t6i sir dung phkrang phap chiet tem protein vai methanol yen cho hieu suet chiet tot (91,96 - 99,20 %) va de lap !pi cao (1,99 - 2,95 %). Phkrang phap dat de nhpy cao (0,1 pg/nril) so vai quy trinh duvc Wei Zhu va Xue-Mei Wang [4]xay dkrng (0,4 pg/ml). Sir dung pha Ong la hen hap dung dich acid acetic pH 2,7 va acetonitril vai tji le 45:55, khong dimg muOi phosphat [3] nen It anh Wang den chat luvng cua cot phan tich khi ap dung trtyc tiep trong nghien ciru dt.rgc Ong hoc, thir ttyang &rang sinh hoc. DO On dinh cua rhein trong huyet twang duvc duy tri trong 60 ngay du thai gian de thtyc hien cac nghien ciru duvc Ong hoc, thin tkrang &rang sinh hoc tren cac che phem chira diacerhein.
Ket luan
Nhirng lieu kien cua phtyang phap HPLC Ong dung de clanh gia rhein trong huyet krang de &vac xey dung. Sir dung phtyang phap chiet tita protein vai not chuen la diclofenac natrị Dieu kien sec kjc tech ten OA C18 (250 x 4,6 mm, 5 pm) vai pha do, IA hen hap acid acetic pH 2,7 va acetonitril vai tjf le 45:55. Phkrang phap dinh krang de duvc them dinh met cach day Tinh tuyen tinh duvc xac dinh trong khoang ruing de 0,1 - 9,5 pg/ml (r2 = 0,9978) vai LLOQ la 0,1 pg/ml. Ket qua de chinh xac, de dung trong ngay va giira cac ngay dpt you ceu cho cho phap phan tich rhein trong huyet twang. Phirang phap c6 the ap dung trong nghien ciru duvc Ong h9c, thin tirang &rang sinh hoc ten cac che phem china diacerhein.
Thi lieu tham khao
1. Geraldine Falgarone, Maxime Dougados (2001), "Diacerein as a disease-modulating agent in (2001), "Diacerein as a disease-modulating agent in
osteoarthritis", Current Rheumatology Reports, 3(6),
479-483.
2. Nicolas P., Tod M., Padoin C., Petitjean 0.
(1998), "Clinical pharmacokinetic of diacerein", Clinical
Pharmacokinetics, 35(5), 547-359.
3. Uday S. Chakrabarty, Uttam Mandal, Uttam Bhaumik, Bappaditya Chatterjee, Animesh Ghosh, Bhaumik, Bappaditya Chatterjee, Animesh Ghosh, Anirban Bose, Tapan K. Pal (2008), "Bioequivalence study of two capsule formulations containing diacerein
50 mg in healthy human subjects", Drug Research,
58(8), 405-409.
4. Wei Zhu, Xue-Mei Wang, Li Zhang, Xiao-Ye Li, Bao-Xiu Wang (2005), "Pharmacokinetic of rhein Li, Bao-Xiu Wang (2005), "Pharmacokinetic of rhein
• Nghien - Ky thuot
in healthy male volunteers following oral and retention enema administration of rhubarb extract: A single dose
studý, The American Journal of Chinese Medicine,
33(6), 839-850.
5. US Department of Health and Human Services, Food and Drug Administration, Centre for Drug
Evalution and Research (1999), Clinical pharmacology
and biopharmaceutics review, Application review:
21-178, Reviewer: Steven B. Johnson, B.S.Pharm, Pharm.D.
6. US Department of Health and Human Services, Food and Drug Administration, Center for Drug
Evaluation and Research (2003), Guidance for Industry-Bioanalytical Method Validation.
(Ngay nhan bai: 25/03/2015 - Ngay duyat clang: 04/09/2015)
Nghien ctiu dinh luving acid valproic
trong huyet tifung ngtrai bang phtrung phap sac ky king khili pho sac ky king khili pho
Phang Chace", T4 Manh Hung', Huang Van Dire, Nguygn Thi Dung', Le Hau3
'Gong ty CO plain Deryc Danaphar, 2 VieM Kiem nghiem thuor Trung trattg Khoa Dtrvc, DH Y Dtectc TP. 1-Pd Chi Minh *E-mail: phungchat@gmail.com
Summary
An ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed for determination of Valproic acid in human plasmạ Valproic acid and the internal standard (Methylparaben) were extracted from human plasma by protein precipitation with acetontril and chromatographed on a C18 column (100 x 2.1 mm, ịd., 1.7 pm) with isocratic elution at a flow rate of 0.3 ml/min, mobile phase of 0.1 % acetic acid - acetontril at a suitable ratiọ The Waters UPLC- MS/MS (Xevo TQD) was operated under the multi reaction-monitoring mode using the electrospray ionization techniquẹ The method was validated over the concentration range of 1.0 to 100 pg/ml. The intra- and inter-day precision and accuracy were within 90 % - 110.0 %. The recovery was 99.6 % — 104.8 %, and 100.5 % for Valproic acid and internal standard, respectivelỵ The method showed well time-saving. The total run time was 2.0 min onlỵ This method seemed useful for BA-BE studies on Valproic acid preparations.
Keywords: Valproic acid, Human plasma, UPLC-MS/MS.
DOA van de
Acid valproic (C811,602, M = 144,211 g/mol) hoc natri valproat (C8H15Na02, M = 166,20 g/mol) la thuoc chong do, ng kinh (dan chat cua acid beo), bi ph5n ly thanh ion valproat b dutrng tiOu h6ạ Tac dung chong dOng kinh cua valproat th6ng qua chAt irc the d5n truyhn thhn kinh la acid gama aminobutyric (GABA) 111. Acid valproic h5p thy quang yhu trong \rung tó ngoei, do v5y de dinh luvng tnyc tier) acid valproic bAng phuvng phap HPLC vbi cac detector thong thubng nhiy UV- VIS, DAD hoc huSinh quang ghn nhu, khOng the
tin hanh duvc, ma can tao d5n xuht cho acid valproic de tao thanh cac hgp chAt help thy UV manh hen, ho5c thanh cac hgp chAt huynh quang [5'7]. tren nguyen ly hoet c1Ong cua phiyang phap sac k9 long — khoi ph6 (LC-MS), cac phutyng phap chiet tach va tham khao tai lieu
[3'4], chOng toi da tin hanh nghien xay dung phuvng phap UPLC-MS/MS co du dO nhey, dec hieu, chinh xac, dinh luting duvc acid valproic trong cac mAu huy5t toting ngutrị Ph(rang phap phan tich se du,gc ap dung trong cac nghien ciru sinh kha dung va twang duvng sinh hoc die phern cht:ra acid valproic.
• Nghien civu - K9 thulat
Nguyen lieu, del tuvng va phúóng phap nghien art, phap nghien art,
Dung mei, hoa chat
Chit chuan
+ Acid vaiproic (AVP) — chuen USP, SKS: VA/11/04, ham lu•gng 99,31%; d6 Am: 0,5%.
+ Methylparaben (MPB) — Vien Kiem nghiem thu6c Trung uvng, SKS: 0213108.02, ham lu,o,ng 99,74%; de Am 0,05%. Methylparaben duvc sir dung lam chat not chuen (IS) trong phtrang phap phan tich.
Dung moi, hoa chat:Dat trail chuan tinh khiet
dung cho HPLC va LC/MS.
Huy& twang Wang khong có AVP: Vien
Hu* hoc N/A truyen mau Trung LK:mg cung cap.
Thiet bi va dieu kien phan tich
Tien hanh phan tich tren may sac ky long kh6i ph6 sieu hieu nang LCMS Waters de duvc hieu chuen theo qui dinh cue ISO/IEC va GLP vai cac dieu kien sac ky va khoi ph6 nhu sau:
Dieu kien sac ky
+ Cot C18; 100 x 2,1 mm; 1,7 pm.
+ Pha d6ng: Acetontril — Acid acetic 0,1%, tY le phO hap. Toc de d6ng 0,3 mL/phut.
+ The tich tient mau: 0,5 pL.
Dieu kien khOi phO
+ Kieu ph6 khoi: SIM el& voi acid vaiproic va MS/MS dei vdei methylparaben, nguon ion hOa: ESI (-);
+ Cac th6ng so cua thiet bi khoi ph6 de phat
hien AVP va MPB nhu, sau:
Chuan bi va xiv ljf cac mau huyet twang china aicd vaiproic
- Chuan bi maw Cac mau huyAt twang (HT)
tv tao chiya AVP duvc chuan bi bang cach h6a tan chuan AVP vao cac mau huyet twang trAng vai cac n6ng d6 khac nhau tir 1 den 100 pg/ml.