As it is well known, CPE efficiency is conditioned by physicochemical
variables as well as the matrix nature of the sample and
analytes [14,18]. Therefore, it is of interest to study the effect of the
experimental physicochemical variables on the analytical response
of the ARV and optimize it towards achieving high sensitivity and
selectivity of the methodology.
Developing analytical method using synthetic solution and
further application on real samples generally requires severe adjustments.
This could lead to a new method different from the
former one. The use of real matrix all along the process is a
convenient choice for characterizing, studying and optimizing the
analytical methodology that is been developed. In this sense, not
only the signal enhancing due to the improvement of the extraction
efficiency resulted significant, but also the matrix load minimization,
which led to a satisfactory chromatographic analysis. These
aspects were what finally determined the sensitivity and selectivity
of the resulting analytical methodology. Thus, those particular
factors of a real matrix, as well as all experimental variables that
condition the analytical responses of the ARV were evaluated,
including extraction pH, surfactant concentration, extraction time
and temperature, salting out effect and the use of modifiers,
including ACN, MeOH and acid, respectively. These studies were
carried out by modifying one variable at the time keeping constant
the remaining ones. A 500 mL aliquot of plasma, originally free of
ARV, was spiked with antiretroviral standard-mix (750 ng mL1
3 TC and ABC, 300 ngmL1 NFV and 150 ngmL1 EFV in H2O:MeOH
75:25 v/v) and 50 mL of Nelfinavir-d3 (Internal Standard,
2200 ng mL1) for carrying out the assays, which were done by
triplicate. The chromatographic peak area was used to evaluate the
impact of modified experimental conditions on the analytical
response.
From Section 3.2e3.5 are presented the results and discussion of
the relevant variables of this work. The remaining variables
mentioned above, did not show significant effect on the analytical
response of the studied ARV.
As it is well known, CPE efficiency is conditioned by physicochemicalvariables as well as the matrix nature of the sample andanalytes [14,18]. Therefore, it is of interest to study the effect of theexperimental physicochemical variables on the analytical responseof the ARV and optimize it towards achieving high sensitivity andselectivity of the methodology.Developing analytical method using synthetic solution andfurther application on real samples generally requires severe adjustments.This could lead to a new method different from theformer one. The use of real matrix all along the process is aconvenient choice for characterizing, studying and optimizing theanalytical methodology that is been developed. In this sense, notonly the signal enhancing due to the improvement of the extractionefficiency resulted significant, but also the matrix load minimization,which led to a satisfactory chromatographic analysis. Theseaspects were what finally determined the sensitivity and selectivityof the resulting analytical methodology. Thus, those particularfactors of a real matrix, as well as all experimental variables thatcondition the analytical responses of the ARV were evaluated,including extraction pH, surfactant concentration, extraction timeand temperature, salting out effect and the use of modifiers,including ACN, MeOH and acid, respectively. These studies werecarried out by modifying one variable at the time keeping constantคนที่เหลือ ส่วนลงตัวขนาด 500 มล.ของพลาสมา เดิมฟรีมี spiked ARV พร้อมด้วยยาต้านไวรัสมาตรฐาน-ผสม (750 ฉบับมล. 13 TC และ ABC, 300 ngmL 1 NFV และ 150 ngmL 1 EFV ใน H2O:MeOH75:25 v/v) และ 50 มล.ของ d3 เนลฟินาเวียร์ (มาตรฐานภายในฉบับที่ 2200 mL 1) สำหรับการดำเนินการ assays ซึ่งทำโดยเพิ่มขึ้นสามเท่า พื้นที่สูงสุดโครมาใช้ในการประเมินการผลกระทบของเงื่อนไขทดลองแก้ไขบนการวิเคราะห์การตอบสนองจาก 3.2e3.5 ส่วนแสดงผลและการอภิปรายตัวแปรที่เกี่ยวข้องของงานนี้ ตัวแปรที่เหลือดังกล่าวข้างต้น ไม่ได้แสดงผลอย่างมีนัยสำคัญที่วิเคราะห์การตอบสนองของ ARV ศึกษา
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