Objective: To establish an HPLC method for the analysis of related substances in near infrared fluorescent dye Cypate and study its photothermal conversion effect in vitro. Methods: The Phenomenex Luna-C18 column (250 mm×4.6 mm, 5 μm) was used for separation with the mobile phase consisting of 0.01 mol·L-1 phosphate buffer (pH 2.5) (A) and acetonitrile (B) in gradient elution model. The gradient elution procedure was as follows:0-2 min, 30%B; 2-25 min, 30%B→52%B; 25-40 min, 52%B→60%B; 40-55 min, 60%B→75%B; 55-60 min, 75%B; 60-61 min, 75%B→30%B; 61-70 min, 30%B. The flow rate was 1.0 mL·min-1, the detection wavelength was 220 nm, and the column temperature was 30 ℃. Results: The minimum resolution between the main peak of Cypate and its adjacent impurity peaks was 1.80(>1.5), and that of other impurities was 1.38(>1.2). There was a good linear relationship between the concentration and the peak area in the range of 0.1-25.1 μg·mL-1(r=0.999 0). The concentration of detection limit and quantitative limit were 0.02 μg·mL-1 and 0.06 μg·mL-1 respectively. In addition, the photothermal conversion effect of Cypate was obviously concentration dependent and time-dependent in 0-210 s. Conclusion: This method meets the requirements of separation between Cypate peak and impurity peaks, and is suitable for the determination of related substances of Cypate. Furthermore, the photothermal conversion effect of Cypate is good.
CAI Liang-liang, HAN Yong, SHEN Kai, WU Wen-yi, ZHU Yong-hong
. Establishment of the method for the detection of related substances in the near infrared fluorescent dye Cypate and its photothermal conversion effect in vitro*[J]. Chinese Journal of Pharmaceutical Analysis, 2021
, 41(10)
: 1757
-1763
.
DOI: 10.16155/j.0254-1793.2021.10.13
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