应用酶解-离子对色谱法测定敷料中透明质酸钠含量

朱翔宇; 唐铭烩; 杨钰娜; 展静

doi:10.12455/j.issn.1671-7104.260211

应用酶解-离子对色谱法测定敷料中透明质酸钠含量

Determination of Sodium Hyaluronate in Dressings by Enzymatic Hydrolysis and Ion-Pair Chromatography

摘要

摘要:
目的建立一种适用于膏状敷料中透明质酸钠含量测定的酶解—离子对高效液相色谱法，以解决基质干扰、操作风险或设备成本较高的问题。
方法样品经30%氯化钠溶液破乳后，用透明质酸酶进行酶解，采用高效液相色谱法检测，参数：Sepax GP-C18色谱柱（4.6×250 mm, 5 μm），以40 mmol/L磷酸二氢钠-10 mmol/L四丁基硫酸氢铵溶液（pH2.7）和乙腈为流动相，流速0.6 mL/min，柱温40 ℃，检测波长232 nm。
结果透明质酸钠在2~100 μg/mL范围内线性关系良好（R²=0.9997），检测限为0.004 mg/g，定量限为0.013 mg/g，重复性RSD为1.34%，中间精密度RSD为2.30%，平均加标回收率在103.20%～106.46%之间。
结论该方法前处理简便、专属性强、精密度、准确度和耐用性良好，可用于膏状敷料中透明质酸钠的含量测定。

Abstract:
Objective To establish an enzymatic hydrolysis-ion-pair high performance liquid chromatography method for the determination of sodium hyaluronate in cream dressings, to address issues such as substrate interference, operational risks, or high equipment costs.
Methods Samples were demulsified with 30% sodium chloride solution, followed by enzymatic hydrolysis by hyaluronidase. The analysis was performed by HPLC under the following parameters: Sepax GP-C18 column (4.6×250 mm, 5 μm); mobile phase consisting of 40 mM sodium dihydrogen phosphate-10 mM tetrabutylammonium hydrogen sulfate solution (pH2.7) and acetonitrile; flow rate of 0.6 mL/min; column temperature of 40℃; and detection wavelength of 232 nm.
Results Sodium hyaluronate has a good linear relationship within the range of 2 to 100 μg/mL (R²= 0.9997). The detection limit (LOD) and quantification limit (LOQ)were of 0.004 mg/g and 0.013 mg/g, respectively. The RSDs for repeatability and intermediate precision were 1.34% and 2.30%, respectively, and the average recoveries of samples ranged from 103.20% to 106.46%.
Conclusion The method is simple in pretreatment, with strong specificity, good precision, accuracy and robustness, and is suitable for the determination of sodium hyaluronate in cream dressings.

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