【商检行业标准(SN)】 进出口化妆品中氢溴酸右美沙芬的测定 液相色谱法

中华人民共和国出入境检验检疫行业标准SN/T 2291—2009
进出口化妆品中氢溴酸右美沙芬的测定液相色谱法
Determination of dextromethorphan hydromide incosmeticsforimportandexport-Liquid chromatographymethod
2009-02-20发布
中华人民共和国
国家质量监督检验检疫总局
2009-09-01实施
本标准的附录A为资料性附录。
本标准由国家认证认可监督管理委员会提出并归口。本标准起草单位：中华人民共和国天津出入境检验检疫局。本标准主要起草人：吴延晖、许泓、林安清、肖亚兵、古珑、张曼、何佳、张骏。本标准系首次发布的出入境检验检疫行业标准SN/T2291—2009
1范围
进出口化妆品中氢溴酸右美沙芬的测定液相色谱法
本标准规定了进出口化妆品中氢漠酸右美沙芬的液相色谱检测方法本标准适用于化妆品中氢漠酸右美沙芬的检测。2原理
SN/T2291—2009
用甲醇提取试样中的氢漠酸右美沙芬，提取液经离心过滤后，用液相色谱-荧光检测器测定，外标法定量。
3试剂和材料
3.1乙睛：色谱纯。
3.2甲醇：色谱纯。
3三乙胺：色谱纯，
3.4冰乙酸：色谱纯
3.53%乙酸溶液：量取30mL冰乙酸，用水定容到1L，用三乙胺调节pH=4.3。3.6
氢溴酸右美沙芬标准物质：纯度≥99%。3.7氢溴酸右美沙芬标准储备溶液：准确称取适量氢溴酸右美沙芬标准物质（精确到0.1mg）.以甲醇配制成浓度为1000μg/mL的标准储备溶液。3.8标准工作溶液：根据标准的灵敏度和仪器线性范围，用甲醇稀释成适当浓度的标准工作液。3.90.22μm有机相滤膜
3.10刻度具塞离心管：15mL。
4仪器
4.1高效液相色谱仪：配有荧光检测器4.2分析天平：感量0.1mg
4.3超声波水浴
4.4冷冻离心机（转速大于3500r/min）。4.5冰箱（-18℃)。
5测定步骤
5.1试样处理
称取试样约1.0g（精确到0.01g），于15mL具塞刻度离心管中，加人甲醇8mL：用力振摇使基质均匀分散后，将溶液定容至10mL，在超声波水浴内超声20min，混勾，在冰箱冷冻室（一18℃或更低温度）放置10min后，于3500r/min下冷冻离心5min，将样品放至室温，取上清液经0.22μm有机滤膜过滤，滤液供液相色谱测定
5.2测定
5.2.1液相色谱条件
5.2.1.1荧光检测器：激发波长280nm，发射波长310nm。5.2.1.2色谱柱：Phenomenex5μmCg250mmX4.6mm.5μm。1
SN/T 2291—2009
5.2.1.3流动相：乙睛+3%乙酸溶液（34+66）。5.2.1.4流速：1.0mL/min。
5.2.1.5柱温：室温。
5.2.1.6进样量：20μL。
5.2.2色谱测定
移取氢溴酸右美沙芬标准储备溶液配制成0.025ug/mL、0.05ug/mL、0.5ug/mL、1.0ug/mL、10μg/mL、20ug/mL标准工作溶液。按色谱条件（5.2.1)进行测定，以色谱峰的峰面积为纵坐标，与其对应的浓度为横坐标作图，绘制标准工作曲线。在上述仪器条件下，氢漠酸右美沙芬的保留时间约为5.4min。标准溶液色谱图参见附录图A.1。试样溶液（5.1)注入液相色谱仪，按色谱条件（5.2.1)进行测定，记录色谱峰的保留时间和峰面积。氢溴酸右美沙芬含量高的试样可取适量用甲醇稀释到线性范围内进行测定。
5.3空白试验
除不加试样外，均按上述操作步骤进行。结果计算
根据标准曲线按式（1)计算：
式中：
X=CXVX1000
mx1000
化妆品中氢溴酸右美沙芬的含量，单位为毫克每千克（mg/kg）：.（1)
从标准工作曲线上查出的试样溶液中氢溴酸右美沙芬的浓度，单位为微克每毫升（ug/mL)；试样定容体积，单位为毫升（mL）；试样的质量，单位为克（g）。
计算结果保留两位小数，计算结果需扣除空白值。7测定低限及回收率
7.1测定低限
本方法对氢溴酸右美沙芬的测定低限为0.5mg/kg。7.2回收率
氢溴酸右美沙芬添加浓度及霜剂、水剂的回收率数据见表1表1氢溴酸右美沙芬添加浓度及霜剂、水剂的回收率试样类型
霜剂类
水剂类
添加浓度/（mg/kg）
回收率范围/%
93.8~102.2
98.7~104.5
99.5~100.6
92.0~102.6
96.2~100.7
93.3~99.2
氢溴酸右美沙芬（5.38min）。
附录A
（资料性附录）
标准品色谱图
氢溴酸右美沙芬标准品液相色谱图9
SN/T2291—2009
SN/T2291—2009
Foreword
AnnexAofthis standardis informativeannex.This standard was proposed by and is under the chargeof National Regulatory Commission for Certi-ficationandAccreditation
This standard was drafted by Tianjin Entry-Exit inspection and Quarantine Bureau of thepeople's Republic of China.
The main drafters of this standard are Wu Yanhui,Xu Hong,Lin Anqing,Xiao Yabing,Gu Long,ZhangMan,HeJia,Zhang Jun
This standard is a professional standard for entry-exit inspection and quarantine promulgated for thefirsttime.
SN/T2291—2009
Determination of dextromethorphan hydromide incosmetics for import and export-Liquid chromatography methodScope
Thisstandard specifiesthedeterminationofdextromethorphanhydromidebyHPLCincosmeticsThis standard is applicable to the determination of dextromethorphan hydromide in cosmetics.2Principle
The dextromethorphan hydromide is extracted with methanol, centrifuged, filtrated and then deter-mined by high performance liquid chromatographywith fluorescencedetection.Makequalitative anal-ysis accordingto external standard method.3Reagentsandmaterials
Acetonitrile:HPLC grade
Methanol:HPLCgrade
Triethlamine:HPLC grade.此内容来自标准下载网
Glacialaceticacid:HPLCgrade
3%Acetic acid solution:30mL Glacialacetic acid,added waterto1L,adjusted pH=4.3by tri.ethylamine
The standard dextromethorphanhydromidePurity≥99%3.6
Thedextromethorphanhydromide standard stock solution:Preciselyweighmodest contentof3.7
standarddextromethorphanhydromide(accurateto0.1mg),preparing1000 μg/mLdextromethor-phanhydromidestandardstocksolutionwithmethanol3.8
The standard working solution:According to the sensitivity and the linear range of instruments.dilutedwithmethanoltoappropriateconcentration.5
SN/T2291—2009
0.22μmorganicphasefilter.
Cypriot scale centrifuge tube:15mL.Apparatus and equipment
HPLc（equippedwithfluorescentdetector).Analytical Balance.
Ultrasonicwaterbathcauldron
Refrigerated centrifuge (>3500 r/min)Refrigerator（-18℃）
Procedure of determination
Sample
Weigh 1. 0 g (accurate to 0. 01 g) of the test sample into 15 mL cypriot scale centrifuge tube, Add8 mL methanol,keep oscillating until the materials fully mixed. Calibrate the volume to 1o mL andthen ultrasound 20 min in ultrasonic water bath cauldron,then shake the tube to make the materialsfully mixed. Put the tube into chilled Room of refrigerator （-18 ℃) for 10 min,3 500 r/min centri-fuge5min,taking the supernatantpassthrough the0.22μm filter filmafter it is back toroomtem-peraturerforHPLCdetermination.5.2
Determination
LC operating conditions
Fluorescencedetection:Excitationwavelength280nm;emitionwavelength310nm.Column：Phenomenex5μmCg.250mmx4.6mm，5μmMobile phase:Acetonitrile +3% glacial acetic acid (adjust pH with triethylamine to 4.3)=34 +Flowrate:1.0mL/min.
Column temperature:room temperature.5.2.1.6
SInjectionvolumn:20μL.
5.2.2LC determination
SN/T2291—2009
Transfer dextromethorphanhydromide standard stock solutionto preparate0.025μg/mL、0.05μg/mL、0.5 μg/mL,1.0 μg/mL,10μg/mL、20 μg/mL standard working curve.Inject20 μL into LC detectorand detect under the (5.2.1) condition.Make the standard curve with peak area as vertical coordi-nate and the corresponding concentration as abscissa. Under the above Lc conditions,the retentiontime of dextromethorphan hydromide is about 5.4 min. Standard grape is shown in annex A. 1.Inject20 μL sample solution(5.1)intoLC detector and detect under the condition(5.2.1),and then recordthe retention time and the peak area of the chromatographic peak. If the content of dextromethor-phan hydromide in the sample is higher than usual,transfer modest content in order to dilute to thelinear range fordetecting.
5.3 Blank test
Followtheupper stepswithouttheadditionof test sample.6
CalculationandexpressionoftheresultCalculate as the following formula(1)according to the standard working curve.C×V×1000
mx1000
Where：
X-thecontentofdextromethorphanhydromideincosmetics,mg/kg;(1）
ctheconcentration of dextromethorphan hydromide in sample solutionfrom the standard workingcurve,μg/mL；
V-thevolumeoftestsampleinthefinalsolution.mL;m-the mass of the test sample,gAfterdiscountingthe effect of blank,preserve two bitof decimal of the results.Limitofdeterminationandrecovery7.1 Limit of determination
The limitof determinationofthismethodof dextromethorphanhydromideis0.5mg/kg7
SN/T 2291—2009
Recovery
The recovery of added concentration of dextromethorphan hydromide in cream,water agent is shownin table 1.
Table1-The recovery of added concentrationofdextromethorphan hydromide in cream,wateragentSampletype
Water agent
added concentration/(mg/kg)
scopeof recovery/%
93.8~102.2
98.7~104.5
99.5~100.6
92.0~102.6
96.2~100.7
93.3~99.2
AnnexA
(lnformativeannex)
Standard chromatography
1-Dextromethorphanhydromide(5.38min)，SN/T2291—2009
Figure A. 1-Standard liquid chromatography of dextromethorphan hydromide9
SN/T 2291-2009
中华人民共和国出人境检验检疫行业标准
进出口化妆品中氢溴酸右美沙芬的测定液相色谱法
SN/T2291—2009
中国标准出版社出版
北京复兴门外三里河北街16号
邮政编码：100045
网址spc.net.cn
电话：6852394668517548
中国标准出版社案皇岛印刷厂印刷开本880×1230
2009年5月第一版
印张1
字数15千字
2009年5月第一次印刷
印数1-2000
书号：155066·2-19674
定价18.00元
小提示：此标准内容仅展示完整标准里的部分截取内容，若需要完整标准请到上方自行免费下载完整标准文档。