LC-MS/MS测定中成药中非法添加物N-乙基他达拉非
|
摘要
目的建立LC-MS/MS定性及定量测定中成药中非法添加N-乙基他达拉非。方法采用DAD检测器进行初步定性筛查,用质谱检测器进行定性确证,再用DAD检测器进行定量。初筛及定量选用SHISEIDO C18柱(250 mm×4.6 mm,5μm),以流动相A[磷酸三乙胺溶液(取三乙胺7 m L,用水稀释至1 000 m L,用磷酸调p H至2.8)-甲醇-乙腈(60∶20∶20)]与流动相B[磷酸三乙胺溶液(取三乙胺7 m L,用水稀释至1 000 m L,用磷酸调p H至2.8)-甲醇-乙腈(8∶46∶46)]梯度洗脱,体积流量1 m L·min-1,检测波长230 nm;确证选用Dikma Spursil C18柱(150 mm×2.1 mm,3μm),以甲醇-乙腈-含0.1%冰乙酸的0.02 mol·L-1乙酸铵溶液(30∶25∶45)等度洗脱,体积流量0.2 m L·min-1。结果 N-乙基他达拉非质量浓度在0.003~0.3 mg·m L-1内与峰面积有良好线性关系,平均回收率为97.8%,RSD为0.7%(n=9);通过对30批次壮阳类中成药进行测定,其中2批检测结果为阳性,含量分别为4.8,4.1 mg·g-1。结论该方法快速、准确、灵敏度高,可作为分析检测壮阳类中成药中非法添加N-乙基他达拉非的有效方法。
OBJECTIVE To establish an HPLC-MS/MS method for the determination of N-ethyl tadalafil which was added illegally into traditional Chinese medicines(TCM). METHODS The experiment steps was as follows: N-ethyl tadalafil was preliminary screened firstly by DAD detector, and then confirmed by mass spectrometer detector and quantitative determined by DAD detector at last. The preliminary screening and qualitative analysis was performed by HPLC-DAD, which was conducted on a SHISEIDO C18 column(250 mm×4.6 mm, 5 μm) with gradient elution at a flow rate of 1 m L·min-1. The mobile phase A consisted of water containing 0.7% triethylamine(adjusted to p H 2.8), methanol and acetonitrile(60∶20∶20); the mobile phase B consisted of water containing 0.7% triethylamine(adjusted to p H 2.8), methanol and acetonitrile(8∶46∶46). The sample was detected at 230 nm. The confirming test was performed by LC-MS/MS, which was conducted on a Dikma Spursil C18 column(150 mm×2.1 mm, 3 μm) at a flow rate of 0.2 m L·min-1 with mobile phase consisted of methanol-acetonitrile-0.02 mol·L-1 ammounium acetate(contained 0.1% acetic acid)(30∶25∶45). RESULTS The linear relationship was good at the range of 0.003-0.3 mg·m L-1, and the average recovery was 97.8%(n=9) with RSD of 0.7%. It was confirmed that N-ethyl tadalafil was added illegally in two batches of sthenic TCM through screening 30 batches of TCM, and the content of N-ethyl tadalafil was 4.8 mg·g-1 and 4.1 mg·g-1, respectively. CONCLUSION The method is fast, accurate and highly sensitive and can be used to screen the illegally sthenic TCM spiked with N-ethyl tadalafil.
OBJECTIVE To establish an HPLC-MS/MS method for the determination of N-ethyl tadalafil which was added illegally into traditional Chinese medicines(TCM). METHODS The experiment steps was as follows: N-ethyl tadalafil was preliminary screened firstly by DAD detector, and then confirmed by mass spectrometer detector and quantitative determined by DAD detector at last. The preliminary screening and qualitative analysis was performed by HPLC-DAD, which was conducted on a SHISEIDO C18 column(250 mm×4.6 mm, 5 μm) with gradient elution at a flow rate of 1 m L·min-1. The mobile phase A consisted of water containing 0.7% triethylamine(adjusted to p H 2.8), methanol and acetonitrile(60∶20∶20); the mobile phase B consisted of water containing 0.7% triethylamine(adjusted to p H 2.8), methanol and acetonitrile(8∶46∶46). The sample was detected at 230 nm. The confirming test was performed by LC-MS/MS, which was conducted on a Dikma Spursil C18 column(150 mm×2.1 mm, 3 μm) at a flow rate of 0.2 m L·min-1 with mobile phase consisted of methanol-acetonitrile-0.02 mol·L-1 ammounium acetate(contained 0.1% acetic acid)(30∶25∶45). RESULTS The linear relationship was good at the range of 0.003-0.3 mg·m L-1, and the average recovery was 97.8%(n=9) with RSD of 0.7%. It was confirmed that N-ethyl tadalafil was added illegally in two batches of sthenic TCM through screening 30 batches of TCM, and the content of N-ethyl tadalafil was 4.8 mg·g-1 and 4.1 mg·g-1, respectively. CONCLUSION The method is fast, accurate and highly sensitive and can be used to screen the illegally sthenic TCM spiked with N-ethyl tadalafil.
引文
[1]郭志鑫,董培智,申国华,等.保健食品非法添加化学药品的特点与监管建议[J].中国药事,2013,27(11):1132-1134.
[2]GUO Z X,DONG P Z,SHEN G H,et al.Characteristics of illegal addition of chemicals to health food and recommendations for supervision[J].Chin Pharm Aff(中国药事),2013,27(11):1132-1134.
[3]ALP M,COSKUN M,GOKER H.Isolation and identification of a new sildenafil analogue adulterated in energy drink:propoxyphenyl sildenafil[J].J Pharm Biomed Anal,2013,72(2):155-158.
[4]HAN K M,LEE J H,HWANG I,et al.Isolation and identification of novel propoxyphenyl thiosildenafil found in natural health food product[J].Food Addit Contam,201431(1):1-6.
[5]GE X,LI L,LOW M Y.Identification of a new sildenafil analogue in a healthy supplement[J].J Pharm Biomed Anal,2011,56(3):491-496.
[6]吴燕,李华龙,米亚娴,等.中药及保健食品中新型PDE-5抑制剂的检测研究[J].药物分析杂志,2016,36(1):117-121.
[7]KEE C L,GE X,KOH H L,et al.Isolation and characterization of propoxyphenyl linked sildenafil and thiosildenafil analogues in health supplements[J].J Pharm Biomed Anal,2012,70(21):265-272.
[8]国家药品监督管理局.2016年第196号公告附件1食品中那非类物质的测定(BJS201601)[S].2016.
[9]ZOU Y H,LIU Y W,ZHENG J.Determination of tetracaine hydrochloride which illegal added in male external products by UHPLC-Q-TOF-MS[J].Chin J Mod Appl Pharm(中国现代应用药学),2017,34(9):1282-1285.
[10]国家药品监督管理局.药品检验补充检验方法和检验项目批准件(2009030)[S].2009.
[11]国家药品监督管理局.食药监食监办28号文附件7缓解体力疲劳类保健食品中非法添加物质检验方[S].2016.
[12]HUANG Z H,CAI D D,CHEN Z Y,et al.Discovery of Nortadalafil in health foods[J].Chin Pharm J(中国药学杂志),2015,50(4):371-374.
[13]董培智,申国华,裴社强,等.降糖类中成药中非法添加新物质格列波脲的确证及裂解规律[J].解放军药学学报,2015,31(6):481-483,488.
[14]SHEN G H,DONG P Z,PEI S Q,et al.Detection of glibornuride illegally added into traditional Chinese medicinal preparations by HPLC-MS-MS[J].Chin Tradit Pat Med(中成药),2014,36(6):1213-1217.
[2]GUO Z X,DONG P Z,SHEN G H,et al.Characteristics of illegal addition of chemicals to health food and recommendations for supervision[J].Chin Pharm Aff(中国药事),2013,27(11):1132-1134.
[3]ALP M,COSKUN M,GOKER H.Isolation and identification of a new sildenafil analogue adulterated in energy drink:propoxyphenyl sildenafil[J].J Pharm Biomed Anal,2013,72(2):155-158.
[4]HAN K M,LEE J H,HWANG I,et al.Isolation and identification of novel propoxyphenyl thiosildenafil found in natural health food product[J].Food Addit Contam,201431(1):1-6.
[5]GE X,LI L,LOW M Y.Identification of a new sildenafil analogue in a healthy supplement[J].J Pharm Biomed Anal,2011,56(3):491-496.
[6]吴燕,李华龙,米亚娴,等.中药及保健食品中新型PDE-5抑制剂的检测研究[J].药物分析杂志,2016,36(1):117-121.
[7]KEE C L,GE X,KOH H L,et al.Isolation and characterization of propoxyphenyl linked sildenafil and thiosildenafil analogues in health supplements[J].J Pharm Biomed Anal,2012,70(21):265-272.
[8]国家药品监督管理局.2016年第196号公告附件1食品中那非类物质的测定(BJS201601)[S].2016.
[9]ZOU Y H,LIU Y W,ZHENG J.Determination of tetracaine hydrochloride which illegal added in male external products by UHPLC-Q-TOF-MS[J].Chin J Mod Appl Pharm(中国现代应用药学),2017,34(9):1282-1285.
[10]国家药品监督管理局.药品检验补充检验方法和检验项目批准件(2009030)[S].2009.
[11]国家药品监督管理局.食药监食监办28号文附件7缓解体力疲劳类保健食品中非法添加物质检验方[S].2016.
[12]HUANG Z H,CAI D D,CHEN Z Y,et al.Discovery of Nortadalafil in health foods[J].Chin Pharm J(中国药学杂志),2015,50(4):371-374.
[13]董培智,申国华,裴社强,等.降糖类中成药中非法添加新物质格列波脲的确证及裂解规律[J].解放军药学学报,2015,31(6):481-483,488.
[14]SHEN G H,DONG P Z,PEI S Q,et al.Detection of glibornuride illegally added into traditional Chinese medicinal preparations by HPLC-MS-MS[J].Chin Tradit Pat Med(中成药),2014,36(6):1213-1217.