應用液相層析串聯式質譜分析法檢出食品膠囊中摻加sildenafil citrate西藥成分之定性定量研究   全文下載

Determination of Sildenafil Citrate Adulterated in a Dietary Supplement Capsule by LC/MS/MS

一種標示由動物器官萃取物製成用於增強性功能之食品膠囊，經以50%甲醇萃取，取濾液直接注入液相層析串聯式質譜儀（LC／MS／MS）作正離子電灑法（ESI+）全質譜掃描（MS 1 scan）分析，質譜圖與本局自建液相層析電腦質譜資料圖庫比對結果，疑似含sildenafil citrate成分。繼而以子代離子掃描（Daughter ion scan），獲得質譜圖與對照標準品比對結果均相符，同時以負離子電灑法（ESI-）證明檸檬酸鹽之存在。檢品溶液經液相層析及配合附光二極體陣列檢出器（photodiode array）檢測，獲得之離子峰滯留時間及紫外光光譜圖均與標準品溶液相符，更確切證實檢品摻加sildenafil citrate西藥成分。為瞭解該食品膠囊中含sildenafil cltrate之量，探討其定量方法而配製系列對照標準品溶液，以pnenzepme作為內部標準品，分離管柱為Cosmosil 5C18-AR, 4.6 × 150 mm；移動相溶媒為甲醇：乙?：1%醋酸（25:17:58），流速：0.5 mL／min，毛細管電壓：3 kV；進樣圓錐口電壓：80 V；碰撞能量：25 eV；離子源溫度：120℃；溶媒揮散溫度：350℃等分析條件，選定親代及子代離子（m／z 475>100）作多重離子裂解監控（MRM, Multiple Reaction Monitoring）定量分析並製作檢量線，線性關係γ2值為0.9977。為探討本分析法之精確度及再現性而進行同日內、異日間之測試與添加回收率試驗，結果其相對標準偏差值分別介於7.56~7.75%及9.09~11.25%之間，添加回收率之相對標準偏差值分別介於3.0~5.0%之間，故本分析法之再現性及精確度均可接受。經探討本分析法之最低檢出極限為40 ng／mL。依上述LC／MS／MS分析法定性定量sildenafil citrate既快速又精確。本案檢體經定量結果含sildenafil citrate為42.8 mg／capsule，對患有心血管疾病正在服用硝酸鹽類心臟藥之患者，在不知情下而服用本檢體，恐有安全之虞。"

A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was used to identify and quantify an adulterated substance, in a dietary supplement capsule. The capsule is claimed to be an extract of animal organs and traditional Chinese herbs and indicated for enhancing sexual activity. At first, the sample was extracted with 50% v/v methanol in water and the extract was injected directly into the LC/MS/MS with no separation and a full scan of positive ion eletrospray (ESI +) analysis was performed. The full scan mass spectrum was compared with the NLFD3/LM library created by Division 3rd of National Laboratories of Foods and Drugs for LC/MS/MS analysis. A sildenafil specific mass spectrum was matched indicating that the capsule may have been adulterated with sildenafil, the active ingredient of VIAGRA. Furthermore, sildenafil was confirmed by carrying out a daughter ion scan and a parent ion scan. The daughter ion spectrum was compared with the NLFD3/LM library. A negative ion eletrospray (ESI –) analysis was performed for confirming the citrate salt in the capsule. The results indicated that the adulterated ingredient was sildenafil citrate. For quantitative analysis of the sildenafil citrate, multiple reaction monitoring (MRM) was performed. Pirenzepine 2HCl was used as the internal standard. A series of standards solution and sample solution with internal standard were analyzed by LC/ MS/MS. The analytical condition is as following: column: Cosmosil 5C18-AR, 4.6 x 150 mm; mobile phase:methanol:acetonitrile:1% acetic acid (25:17:58); capillary voltage: 3 kV;cone voltage: 80 V; collision energy: 25 eV; source temperature: 120°C; desolvation temperature: 350°C. The detection limit was 40 ng/mL. An excellent calibration curve of sildenafil citrate was obtained with the γ2 correlation coefficient of 0.9977. The RSDs of interday and intraday were between 7.56~7.75% and 9.09~11.25%, respectively. The analytical strategy and method is suitable for identification and measurement of sildenafil citrate adulterated in capsule. Amount of sildenafil citrate in the capsule was determined as 42.8 mg/capsule. 一種標示由動物器官萃取物製成用於增強性功能之食品膠囊，經以50%甲醇萃取，取濾液直接注入液相層析串聯式質譜儀（LC／MS／MS）作正離子電灑法（ESI+）全質譜掃描（MS 1 scan）分析，質譜圖與本局自建液相層析電腦質譜資料圖庫比對結果，疑似含sildenafil citrate成分。繼而以子代離子掃描（Daughter ion scan），獲得質譜圖與對照標準品比對結果均相符，同時以負離子電灑法（ESI-）證明檸檬酸鹽之存在。檢品溶液經液相層析及配合附光二極體陣列檢出器（photodiode array）檢測，獲得之離子峰滯留時間及紫外光光譜圖均與標準品溶液相符，更確切證實檢品摻加sildenafil citrate西藥成分。為瞭解該食品膠囊中含sildenafil cltrate之量，探討其定量方法而配製系列對照標準品溶液，以pnenzepme作為內部標準品，分離管柱為Cosmosil 5C18-AR, 4.6 × 150 mm；移動相溶媒為甲醇：乙腈：1%醋酸（25:17:58），流速：0.5 mL／min，毛細管電壓：3 kV；進樣圓錐口電壓：80 V；碰撞能量：25 eV；離子源溫度：120℃；溶媒揮散溫度：350℃等分析條件，選定親代及子代離子（m／z 475>100）作多重離子裂解監控（MRM, Multiple Reaction Monitoring）定量分析並製作檢量線，線性關係γ2值為0.9977。為探討本分析法之精確度及再現性而進行同日內、異日間之測試與添加回收率試驗，結果其相對標準偏差值分別介於7.56～7.75%及9.09～11.25%之間，添加回收率之相對標準偏差值分別介於3.0～5.0%之間，故本分析法之再現性及精確度均可接受。經探討本分析法之最低檢出極限為40 ng／mL。依上述LC／MS／MS分析法定性定量sildenafil citrate既快速又精確。本案檢體經定量結果含sildenafil citrate為42.8 mg／capsule，對患有心血管疾病正在服用硝酸鹽類心臟藥之患者，在不知情下而服用本檢體，恐有安全之虞。