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Linearity of calibration curves, LLOD and LLOQ
       LLOQ can be quantitated with accuracy within 15% of nominal and precision not exceeding 15% CV, and was 4.0 ng/mL for TB-II and 3.9 ng/mL for TA-III. LLOD of TB-II and TA-III was obtained when the solution was diluted to 1.8 ng/mL and 1.6 ng/mL, respectively, and the signal-to-noise ratio was 2-fold. The calibration curve of TB-II was y = 0.3379x + 0.02973 (r = 0.9978) and was y = 1.778x−0.02613(r = 0.9989) for TA-III. The difference between the nominal standard concentration and the back-calculated concentration from the weighed linear regression line was varied from−4.3% to 4.6% for each point on the standard curve (CV varied from 1.7% to 8.2%).
  Carry-over was tested by injecting two processed blank matrix samples subsequently after injection of an ULOQ (the upper limit of quantification) sample in three independent runs.The response in the first blank matrix at the retention time of the analytes should be less than 20% of the mean response of a LLOQ sample for the
analytes. The response in the first blank matrix at the retention time of the analytes was less than 7% of the mean response at the LLOQ for TA-III and 3% for TB-II. Therefore, the carry-over test was found to be acceptable.

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