Desarrollo y validación de un método analítico para la cuantificación de Ranitidina en plasma por Cromatografía Líquida de Alta Resolución (HPLC)
Abstract
A reversed phase high - liquid chromatography method for quantifying ranitidine using nizatidine like a internal standard has been developed. The method first involves a purification step of samples using a solid - phase extraction with C18 columns. The extracts have been evaporated at 60 oC in a water bath under a stream of nitrogen gas. The quantitation has been carried out onto C8 column, with a mobile phase of ammonium phosphate pH 7.8 - methanol (70:30, v/v), at a flow rate of 1 mL/min and a detection at a wavelength of 322 nm. The procedure has been validated and linear responses were obtained between 10.2 and 1600 ng/mL. Other parameters of validation such as: selectivity, precision, accuracy and limit of quantitation were in satisfactory agreement with the Report Conference from Washington. The stability of ranitidine in plasma has been studied when the samples were subjected to cycles of freezing and thawing and during the storage period.
Full text
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' CD7 ( +5 (" 1-!+( +7+0,( C( +7)"1,( /+"0(# +(1)+?1+!0++5 ; 1-!$!+(73+1 /)/+1 (1"( !()/ 7 1 1)!1+!0 1-!+!1 . C+!+( /+*"!)/+1 (1"( !1)!()/%2 0)/),0+!)/ 7 1 1)!1+!0 1-!54 /+( 0 ,( %$); )/()/),0+!)//)!!?+)+/ (%A$:S 7)()6"++(340))/ 0/? 1+()/10+)/+ L /B!*0)!GC (L)E/B)71+(+, )+! (!*0)!+!+!+)+2:::K+!()+?++!0+ 7+1/-!$ +I 100"/++5 (N 1)!()/3/3)/ 0)/ 1)!()/6"+/++0+3! ( 7+1/-!$)!( 1)!1+!0 1-!3+ ),0+! 7 1 1)!; 1+!0 1-!/+1 (1"( +(7)1+!0 O+++/5 ; 1-!+(5 ()+ ($4 /+( 0 ,( %$)/5 ()+/ ),0+!)/0 3,4!1"37(+!1)!()/10+)/+ L /B!*0)!G;K7 ( +I 100"+340))/ ,) ! (.01)/T%A$:S$ /1)!1+!0 1)!+/),0+! /7 1 "! +( /3"+/0 / ! (# /1)!+(?!++5 (" +((.30++1" !0?1 1-!'+!!*P3'/)!=K$2' %:$@C%%$8$(+/+%%$9SC+(7)1+!0 O+ ++/5 1-!+(5 ()+ (+/+2$8S= )!; ?++!1 +L /B!*0)!G1)!/+ 6"++!+((.; 30++1" !0?1 1-!/++,+!),0+!+!; ?+)+/ (2:$:SC+/5 1)!+/+(5 ()+ ( +!0+T2:$:S$ )/+/"(0 )/),0+!)/+!+(+/0")++/; 0 ,( , O)11()/+1)!*+( 1-!C+/1)!*+; ( 1-!/+7+/+!0 !+!( 0 ,( 2$ +0+3! /()/5 ()+/3+)/+1)!; 1+!0 1-!),0+!)/1)!1 "! +( /1)!; 1+!0 1)!+/" !0+()/11()/+1)!*+( 1-! C+/1)!*+( 1-!/)!?++!0+/+( 1)!1+!; 0 1-!+( /)("1-!)*! (/+1)!/0"C++( !0+5 ()+1)!? !# 7 ( /%2+0+3! ; 1)!+/'+1 1)!1+!0 1-!7 0-!'+(+!/ ; C)+7+1/-!C+I 100"@4 /+( 0 ,( 2$ 1&#+/"(0 )/+(+/0")+7+1/-!C+I 100" Concentración real (ng/mL) Concentración medida (ng/mL) Día 1 Día 2 Día 3 Día 4 63.8 59.6 56.7 56.2 58.0 55.3 56.7 66.6 62.0 61.6 71.7 58.3 59.2 C.V reprodu.: 7.95 %D: 5.68 C.V repetibilidad 3.19 2.40 4.34 11.85 398.6 457.1 402.5 437.9 426.7 535.6 425.2 412.6 447.8 411.9 439.5 459.7 453.7 C.V reprodu.: 4.33 %D: 8.92 C.V repetibilidad 6.40 1.31 4.83 2.30 1195.8 1335.7 1338.9 1386.8 1306.5 1349.8 1357.1 1306.9 1396.1 1345.7 1369.6 1329.5 1326.6 C.V reprodu.: 1.90 %D: 12.91 C.V repetibilidad 2.11 2.04 1.89 1.79
J "2#)!1+!0 1)!+/+ !0! !*P3" !0++(+/0")++/0 ,( , O)11()/+1)!*+( 1-!C+/1)!; *+( 1-!C !F(//+/0 ./01)+()/ 0)/+(+/0")+7+1/-!C+I 100"$ &$=+/5 1-!+/0F! 63.8 58.1 68.1 67.5 60.2 4.8 49.5 – 70.5 398.7 429.8 418.7 444.3 434.2 18.8 392.8 – 475.5 1195.8 1337.3 1552.5 1285.0 1350.3 25.7 1293.8 – 1406.7 Estudio de estabilidad ciclos de congelación y descongelación Ensayo de exactitud y precisión Concentración real (ng/mL) Día 1 (ng/mL) Día 3 (ng/mL) Día 4 (ng/mL) Media (ng/mL) D.S.* Intervalo de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studio de estabilidad en condiciones de almacenamiento Análisis estadístico Concentración Día 0 (ng/mL) Día 7 (ng/mL) Día 21 (ng/mL) Día 35 (ng/mL) intercepto pendiente Intervalo del 95 % para la pendiente Alta 3529.3 * 3553.2 3567.1 3529.6 0.5065 0.60 – 1.57 Media 2061.9 2013.7 2000.9 2027.9 2038.1 -0.7653 -5.36 – 3.83 Baja 19.1 25.6 30.3 22.0 20.3 1.8055 -0.05 – 1.12 & /3"+/0 //+7++)!7) 11+!0+ $$$
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