固相萃取柱净化-超高效液相色谱-串联质谱法测定助眠类保健品中22种禁用安神类药物的含量

应用超高效液相色谱-串联质谱法测定了助眠类保健品中22种禁用安神类药物的含量。将样品连同其包衣和胶囊壳一起研碎后称取1.000g样品用甲醇溶解,经涡旋混匀并超声15min,用甲醇定容至25.0mL。离心后取上清液5.00mL,经Oasis PRiME HLB固相萃取柱净化处理,收集经净化的流出液,与用于淋洗固相萃取柱的甲醇2mL合并,于40℃下吹氮至溶液近干。用体积比为1∶1的0.05%(体积分数,下同)甲酸-甲醇混合溶液1.00mL溶解残渣,所得溶液经过0.22μm滤膜过滤,滤液供色谱分离。选用Poroshell 120EC-C18色谱柱(50mm×3.0mm,2.7μm)作为固定相,用不同比例的(A)含0.05%甲酸的5mmol·L^-1乙酸铵溶液和(B)乙腈的混合溶液作为流动相按程序进行梯度洗脱。质谱测定中采用电喷雾离子源和多反应监测模式。结果表明:22种禁用安神类药物的标准曲线中有17种线性范围为4~800μg·L^-1,有1种为40~8 000μg·L^-1,还有4种为400~80 000μg·L^-1,其检出限(3S/N)为0.02~2μg·g-1。用标准加入法进行回收试验,测得回收率为81.2%~98.9%,测定值的相对标准偏差(n=6)为3.1%~9.7%。

UPLC-MS/MS Determination of 22 Forbidden Sedative-Hypnotics in Sleep-Aiding Health Care Products with SPE Column Purification

UPLC-MS/MS was applied to the determination of 22 forbidden sedative-hypnotics in some sleepaiding health care products.Sample for analysis was prepared by grinding the health care food itself together with its capsula into powder and a portion of 1.000 g was taken and dissolved in methanol with aid of ultrasonic treatment for 15 min.The solution was diluted to 25.0 mL with methanol and centrifuged.An aliquot of 5.00 mL of the supernatant was taken and purified by passing through the Oasis PRiME HLB SPE column.The eluate from the SPE column was collected and mixed with the methanol(2 mL)used in washing the column.The mixed solution was then evaporated to neardryness by N2-blowing at 40 ℃.The residue was taken up with 1.00 mL of a mixed solution of 0.005%(volume fraction,same as below)formic acid-methanol with the volume ratio of 1∶1,and the solution was filtered through 0.22μm filtering membrane and the filtrate was used for UPLC separation by passing through Poroshell 120 EC-C18 chromatographic column(50 mm×3.0 mm,2.7μm)as stationary phase,and mixture of(A)5 mmol·L^-1 NH4OAc solution(containting 0.05% formic acid)and(B)acetonitrile in various ratios were used as mobile phases in gradient elution.In MS/MS determination,ESI and MRM mode were adopted.It was shown that among the standard curves of the 22 forbidden sedative-hypnotics,linearity ranges for 17 sedativehypnotics were same between 4μg·L^-1 and 800μg·L^-1,for one sedative-hypnotic between 40μg·L^-1 and8 000μg·L^-1 and for the remaider 4 sedative-hypnotics between 400μg·L-1 and 80 000μg·L^-1;the detection limits(3 S/N)found for these sedative-hypnotics were in the range of 0.02-2μg·g^-1.Recovery was tested by standard addition method,giving values of recovery in the range of 81.2%-98.9%,and values of RSDs(n=6)in the range of 3.1%-9.7%.