超高效液相色谱-三重四极杆质谱法用于阿胶、龟甲胶、鹿角胶中猪皮源成分的检测

目的：建立超高效液相色谱-三重四极杆质谱法（UPLC-QQQ/MS法）测定阿胶、龟甲胶和鹿角胶中猪皮源成分。方法：采用胰蛋白酶进行酶解处理，色谱柱为ZORBAX SB-C₁₈（2.1 mm×150 mm，5 μm）；以0.1%甲酸溶液（A）-乙腈（B）为流动相，梯度洗脱（0～25 min，95%A→80%A；25～40 min，80%A→50%A)，流速为0.3 mL·min-1；电喷雾离子源（ESI），正离子模式下扫描，采用多重反应监测模式（MRM），对特征肽目标离子m/z 774.5（双电荷）→977.8和m/z 774.5（双电荷）→1 034.6进行检测。结果：阿胶中新阿胶掺入量为1%～12%、龟甲胶中新阿胶掺入量为2%～11%、鹿角胶中新阿胶掺入量为3%～12%的范围内，掺入量与色谱峰面积成正比，线性方程分别为Y=4 471X-29.00（r²=0.997 1），Y=4 468X-63.33（r²=0.996 3），Y=4 262X-74.48（r²=0.998 0）。检测下限浓度为0.1 g·mL-1。5批阿胶、5批龟甲胶、5批鹿角胶中，有1批阿胶、1批龟甲胶、2批鹿角胶可疑样品中检出新阿胶。结论：所建立方法操作简便，专属性强，灵敏度高，重复性良好，可用于阿胶、龟甲胶和鹿角胶中掺伪成分猪皮源的检测。

Objective: To establish a ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-QQQ/MS) method for the identification of ingredients from pig skin in donkey-hide gelatin,tortoise-shell glue and deer-horn glue. Methods: Trypsin was used for enzymatic hydrolysis. UPLC-QQQ/MS was used to detect ingredients from pig skin in donkey-hide gelatins,tortoise-shell glue and deer-horn glue. Determination was carried out by means of an Agilent SB-C₁₈(2.1 mm×150 mm,1.8 μm) column. The mobile phase was composed of water solvent containing 0.1% formic acid(A)and acetonitrile(B)with gradient elution(0-25 min,95%A→80%A;25-40 min,80%A→50%A)at the flow rate of 0.3 mL·min^-1. The electrospray ionization(ESI)source was performed in multiple reaction monitoring(MRM)mode of the transitions of m/z 774.5(double-charge)→977.8 and m/z 774.5(double-charge)→1 034.6. Results: The adulteration rate was proportional to the chromatographic peak area when the adulteration pig-hide gelatin was 1%-12% in donkey-hide gelatin,2%-11% in tortoise-shell glue and 3%-12% in deer-horn glue,respectively. The linear equations were Y=4 471X-29.00(R²=0.997 1),Y=4 468X-63.33(R²=0.996 3),and Y=4 262X-74.48(R²=0.998 0),respectively. The lower limit of detection concentration of pig-hide gelatin was 0.1 g·mL^-1. Ingredients from pig skin can be detected in one of fives batches of donkey-hide gelatin,one of fives batches of tortoise-hide gelatin and two of fives batches of deer-horn gelatin. Conclusion: The established method is simple,specific,sensitive and repetitive. It can be used for identification of ingredients from pig skin in donkey-hide gelatin,tortoise-shell glue and deer-horn glue.

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