The limit of detection (LOD) and the limit of quantification (LOQ)
were calculated according to the 3sb/m and 10sb/m criteria, where sb
is the standard deviation of peak area obtained after 7 consecutive analysis
of the lower analyte concentration solution and m was the slope of
the regression line. The LOD and LOQ obtained are presented in Table 2.
Repeatability lower than 2.0% was estimated using the relative
standard deviation (RSD) based on three consecutive measurements
of three concentration levels (5.0; 15.0; and 40.0 μg mL−1
) of strontium
ranelate and aspartame standard solutions under the same experimental
conditions.
The intermediate precision of the method was demonstrated by
three replicate measurements of aspartame and strontium ranelate
solution (15 μg mL−1
), made by the same analyst on different days.
The Student's t-test (confidence limit of 95%) was used to compare
the averages of the areas obtained on different days. The calculated
values of t (taspartame = 1.329 and tstrontium ranelate = 0.364) were lesser
than the tabulated value (2.780), indicating that the developed CZE
method presents good intermediate precision [34].
The robustness of the method was evaluated taking into consideration
the influence of the pH variation of the BGE in the peaks areas of
aspartame and of strontium ranelate (in the form of anion ranelate). A
robust condition with no significant area variation was found for the
pH range from 9.2 to 9.6. The F values were calculated based on
ANOVA. According to the results, the calculated F values for this
parameter (Faspartame = 5.3 and Fstrontium ranelate = 1.7) did not exceed
the tabulated value (7.388), indicating no statistical difference between
the means obtained at different pH values.
The method accuracy was defined according to the triplicate measurement
of three different concentration levels of analytes (5.0, 15.0,
and 40.0 μg mL−1
). To evaluate this parameter, the Student's t-test
(confidence limit of 95%) was applied. It showed that the calculated
values of t for each concentration (aspartame: t5.0 = 3.46; t15.0 = 1.44;
t40.0 = 1.65/strontium ranelate: t5.0 = 1.91; t15.0 = 0.17; t40.0 = 0.04)
were all lower than the tabulated value (4.30). This proved that the
developed method is accurate because there is no statistical difference
between the experimental concentrations and theoretical
concentrations.
3.4. Method application
The feasibility of the method (CZE) was verified by recovery tests in
pharmaceutical formulation sample (the composition as described in
Section 2.3), according to the guidance on validation of analytical
methods of Brazilian National Institute of Metrology, Quality and
Technology [35]. The recovery of strontium ranelate and aspartame was
estimated using different sample solutions spiked with known amounts
of analytes at three different levels (5.0; 15.0 and 30.0 μg mL−1
).
Non-spiked samples were run in parallel. Tests were performed in
three replicates and the recovery was calculated using the following
equation: Recovery = [(C1 − C2)/C3] ∗ 100, where C1 is the concentration
of the analytes in the fortified sample; C2 is the concentration
of the analyte in the sample unfortified and C3 is the concentration of
analyte added to the spiked sample. Average recoveries between 85
and 111% were obtained and the results are shown in Table 3.
The absorption spectra of electrophoretic peaks of the sample were
monitorated simultaneously during the analysis of the pharmaceutical
sample without the addition of strontium ranelate and aspartame standards.
Identification of the strontium ranelate and aspartame peaks was
confirmed by wavelength of maximum absorption at 235 nm and
198 nm, respectively, which demonstrated the ability and selectivity
of the method to identify the analytes of interest in the pharmaceutical
formulation, even in the presence of other substances (maltodextrin
and mannitol) in the sample matrix.
The limit of detection (LOD) and the limit of quantification (LOQ)were calculated according to the 3sb/m and 10sb/m criteria, where sbis the standard deviation of peak area obtained after 7 consecutive analysisof the lower analyte concentration solution and m was the slope ofthe regression line. The LOD and LOQ obtained are presented in Table 2.Repeatability lower than 2.0% was estimated using the relativestandard deviation (RSD) based on three consecutive measurementsof three concentration levels (5.0; 15.0; and 40.0 μg mL−1) of strontiumranelate and aspartame standard solutions under the same experimentalconditions.The intermediate precision of the method was demonstrated bythree replicate measurements of aspartame and strontium ranelatesolution (15 μg mL−1), made by the same analyst on different days.The Student's t-test (confidence limit of 95%) was used to comparethe averages of the areas obtained on different days. The calculatedvalues of t (taspartame = 1.329 and tstrontium ranelate = 0.364) were lesserthan the tabulated value (2.780), indicating that the developed CZEmethod presents good intermediate precision [34].The robustness of the method was evaluated taking into considerationthe influence of the pH variation of the BGE in the peaks areas ofaspartame and of strontium ranelate (in the form of anion ranelate). Arobust condition with no significant area variation was found for thepH range from 9.2 to 9.6. The F values were calculated based onANOVA. According to the results, the calculated F values for thisparameter (Faspartame = 5.3 and Fstrontium ranelate = 1.7) did not exceedthe tabulated value (7.388), indicating no statistical difference betweenthe means obtained at different pH values.The method accuracy was defined according to the triplicate measurementof three different concentration levels of analytes (5.0, 15.0,and 40.0 μg mL−1). To evaluate this parameter, the Student's t-test(confidence limit of 95%) was applied. It showed that the calculatedvalues of t for each concentration (aspartame: t5.0 = 3.46; t15.0 = 1.44;t40.0 = 1.65/strontium ranelate: t5.0 = 1.91; t15.0 = 0.17; t40.0 = 0.04)were all lower than the tabulated value (4.30). This proved that thedeveloped method is accurate because there is no statistical differencebetween the experimental concentrations and theoreticalconcentrations.3.4. Method applicationThe feasibility of the method (CZE) was verified by recovery tests inpharmaceutical formulation sample (the composition as described inSection 2.3), according to the guidance on validation of analyticalmethods of Brazilian National Institute of Metrology, Quality andTechnology [35]. The recovery of strontium ranelate and aspartame wasestimated using different sample solutions spiked with known amountsof analytes at three different levels (5.0; 15.0 and 30.0 μg mL−1).Non-spiked samples were run in parallel. Tests were performed inthree replicates and the recovery was calculated using the followingequation: Recovery = [(C1 − C2)/C3] ∗ 100, where C1 is the concentrationof the analytes in the fortified sample; C2 is the concentrationof the analyte in the sample unfortified and C3 is the concentration ofanalyte added to the spiked sample. Average recoveries between 85and 111% were obtained and the results are shown in Table 3.The absorption spectra of electrophoretic peaks of the sample weremonitorated simultaneously during the analysis of the pharmaceuticalsample without the addition of strontium ranelate and aspartame standards.Identification of the strontium ranelate and aspartame peaks wasconfirmed by wavelength of maximum absorption at 235 nm and198 nm, respectively, which demonstrated the ability and selectivityof the method to identify the analytes of interest in the pharmaceuticalformulation, even in the presence of other substances (maltodextrinand mannitol) in the sample matrix.
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The limit of detection (LOD) and the limit of quantification (LOQ)
were calculated according to the 3sb/m and 10sb/m criteria, where sb
is the standard deviation of peak area obtained after 7 consecutive analysis
of the lower analyte concentration solution and m was the slope of
the regression line. The LOD and LOQ obtained are presented in Table 2.
Repeatability lower than 2.0% was estimated using the relative
standard deviation (RSD) based on three consecutive measurements
of three concentration levels (5.0; 15.0; and 40.0 μg mL−1
) of strontium
ranelate and aspartame standard solutions under the same experimental
conditions.
The intermediate precision of the method was demonstrated by
three replicate measurements of aspartame and strontium ranelate
วิธีแก้ปัญหา ( 15 μกรัมต่อมิลลิลิตร− 1
) ทำโดยนักวิเคราะห์เดียวกันในวันที่แตกต่างกัน .
ของนักเรียน ) ( ความเชื่อมั่น 95% ) คือใช้ในการเปรียบเทียบค่าเฉลี่ยของพื้นที่
ได้ในวันที่แตกต่างกัน คำนวณค่า T
taspartame = 1.329 tstrontium = 0.364 และ ranelate ) น้อยกว่า
กว่าตารางค่า ( 2.780 ) แสดงว่าพัฒนา CYP
วิธีการนำเสนอที่ดี แม่นยำปานกลาง [ 34 ] .
ทนทานของวิธีการประเมินพิจารณา
อิทธิพลของ pH เปลี่ยนแปลงของ bge ในหุบด้าน
สารให้ความหวานและสทรอนเทียม ranelate ( ในรูปแบบของ ranelate ไอออน ) เป็นเงื่อนไขที่แข็งแกร่งกับพื้นที่สำคัญ
ไม่มีการพบ
pH ช่วงจาก 9.2 ถึง 9.6 . คำนวณตามค่า F
ANOVA According to the results, the calculated F values for this
parameter (Faspartame = 5.3 and Fstrontium ranelate = 1.7) did not exceed
the tabulated value (7.388), indicating no statistical difference between
the means obtained at different pH values.
The method accuracy was defined according to the triplicate measurement
of three different concentration levels of analytes (5.0, 15.0,
and 40.0 μg mL−1
). To evaluate this parameter, the Student's t-test
(confidence limit of 95%) was applied. It showed that the calculated
values of t for each concentration (aspartame: t5.0 = 3.46; t15.0 = 1.44;
t40.0 = 1.65/strontium ranelate: t5.0 = 1.91; t15.0 = 0.17; t40.0 = 0.04)
were all lower than the tabulated value (4.30). This proved that the
การพัฒนาวิธีที่ถูกต้อง เพราะไม่มีความแตกต่างทางสถิติระหว่าง ความเข้มข้นและปริมาณ
ทดลองทฤษฎี
.
3.4 . วิธีการสมัคร
ความเป็นไปได้ของวิธีการ ( CYP ) คือการยืนยันโดยการทดสอบใน
เภสัชกรรม formulation ตัวอย่างการกู้คืน ( องค์ประกอบตามที่อธิบายไว้ในส่วน 2.3
) ตามคำแนะนำในการตรวจสอบวิเคราะห์
วิธีการของสถาบันมาตรวิทยาแห่งชาติบราซิล คุณภาพและ
เทคโนโลยี [ 3 ] การกู้คืนของ ranelate อัสซีเรียและสารให้ความหวานคือ
ประมาณโดยใช้โซลูชั่นตัวอย่างต่างๆถูกแทงด้วยปริมาณของสารที่รู้จักกัน
3 ระดับ ( 5.0 กรัมต่อมิลลิลิตร ; 15.0 และส่งเสริมμ− 1
)
โนนหนามจำนวนวิ่งขนาน ทดสอบใน
3 ซ้ำและการกู้คืนถูกคำนวณโดยใช้สมการต่อไปนี้
: การกู้คืน = [ ( C1 C2 C3 ] ∗− ) / 100 ที่ C1 เป็นสมาธิ
ของสารในการเก็บตัวอย่าง ; C2 เป็นสมาธิ
ของครูในตัวอย่างซีและ C3 คือความเข้มข้นของ
ครูเพิ่มไป ตัวอย่างที่ได้ถูกแทง อุณหภูมิเฉลี่ยระหว่าง 85
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