有關(guān)物質(zhì)方法確認(rèn)方案

1、METHOD VERIFICATION PROTOCOL FORCHROMATOGRAPHIC PURITY OF CYTARABINE阿糖胞苷有關(guān)物質(zhì)檢查方法確認(rèn)方案Area to be Distributed :AD, QA分發(fā)部門：分析部，質(zhì)量保證部Print Name:姓名：Title:職位：Sig nature:簽名：Date:日期：Author:起草人：Approver: 批準(zhǔn)人：Approver: 批準(zhǔn)人：Reviewer:審核人：Approver: 批準(zhǔn)人：Contents目錄一、Introduction3二、Scope3三、Responsibility3四、Definitio

2、ns3五、Project5六、References25Introduction簡介# is in the process of developing Cytarabine Injection, a liquid product for parenteral adm ini strati on, for #. Cytarabi ne is a chemotherapy age nt with a molecular weight of 2432 The API from Zhejiang Hisun Pharmaceutical Co. Ltd, which is approved by FDA

3、, is used for Cytarabine Injection by Lummy.The Chromatographic purity method for Cytarabine is the method in the current USP34. The HPLC method for Chromatographic purity test of Cytarabine is a quantitative method. The method will be verified according to USP<1226>, including system suitabil

4、ity, specificity, LOD, LOQ, precision, solution stability and mobile phase stability .阿糖胞苷注射液是一種臨床用液體產(chǎn)品，#藥業(yè)為#藥業(yè)研發(fā)該品種。阿糖胞苷是一種化學(xué)藥，分子量為 243.2。萊美研發(fā)阿糖胞苷注射液所用原料藥來源于浙江海正 藥業(yè)，是獲得了 FDA認(rèn)證的原料藥。阿糖胞苷色譜純度檢測方法來自現(xiàn)行USP34,色譜純度檢查方法，是定量檢測方法。依照USP<1226>，本次確認(rèn)內(nèi)容包括系統(tǒng)適用性，專屬性，檢測限，定量限、精密度、溶液穩(wěn)定性和流動相穩(wěn)定性。Scope范圍This protoco

5、l applies to the verification for chromatographic purity of cytarabine.該方案適用于阿糖胞苷色譜純度的方法確認(rèn)。Responsibility職責(zé)四、Definitions定義NameDepartme ntTitleResp on sibility姓名所在部門職稱/職務(wù)職責(zé)Group leader, responsible for approve of protocols, records and reports, and organize the implementation of the protocol.組長，負(fù)責(zé)方案、

6、記錄和報告的審查 批準(zhǔn)并組織方案的實施。Initiate and implement the protocol 起草并執(zhí)行方案Approve protocols, records and reports 方案、記錄和報告的審查批準(zhǔn)Monitor the implementation of the protocol監(jiān)督確認(rèn)方案的實施1 Limit of Detectio n (LOD)檢測限(LOD)The limit of detecti on of an in dividual an alytical procedure is the lowest amount of an alyte in

7、 a sample which can be detected but not n ecessarily qua ntitated as an exact value. 分析規(guī)程中的檢測限是指樣品中的被分析物在該分析方法中能被檢測，但不需要準(zhǔn) 確定量的最低濃度。The detect ion limit is determ ined by an alyz ing samples with known concen trati ons of analyte and by establishing the minimum level at which the analyte can be relia

8、bly detected.檢測限是通過分析樣品已知濃度的被分析物，從而建立其可穩(wěn)定檢測的最低濃度水 平。2 Limit of Qua ntitatio n (LOQ)定量限(LOQ)The qua ntitati on limit of an in dividual an alytical procedure is the lowest amount of analyte in a sample which can be quantitatively determined with suitable precision and accuracy.分析方法中的定量限是指樣品中被分析物能夠被定量檢

9、測并保證一定精密度和準(zhǔn)確 度的最低濃度。The limit of qua ntitati on is a parameter of qua ntitative assays for low levels of compo unds in sample matrices, and is used particularly for the determ in ati on of impurities an d/or degradati on products.定量限是在樣品中低濃度化合物被定量檢測的參數(shù)，該參數(shù)經(jīng)常被用于雜質(zhì)和/或降解產(chǎn)物的測定。3 system suitability系統(tǒng)適用性S

10、ystem suitability tests are an integral part of gas and liquid chromatographic methods. They are used to verify that resoluti on and reproducibility of the chromatographic system are adequate for the analysis to be done. The tests are based on the concept that the equipment, electronics, analytical

11、operations, and sample to be analyzed constitute an in tegral system that can be evaluated as such.系統(tǒng)適用性試驗是氣相色譜和液相色譜方法的必要組成部。用以確認(rèn)該分析方法色 譜系統(tǒng)的分離度和重現(xiàn)性是適用的。該試驗把分析設(shè)備、電子儀器、分析操作和分 析樣品作為一個整體來進(jìn)行評估。4 Specificity專屬性Specificity is the ability to assess the analyte unequivocally in the presenee of components whi

12、ch may be expected to be prese nt. Typically these might in clude process impurities, degradates, matrix, etc.專屬性是指在一些預(yù)期組分的存在下，被分析物被明確檢測評估的能力。通常這些 預(yù)期組分是指工藝雜質(zhì)，降解產(chǎn)物，基質(zhì)等。5 Precisi on精密度The precisi on of an an alytical procedure expresses the close ness of agreeme nt (degree of scatter) betwee n a serie

13、s of measureme nts obta ined from multiple sampli ng of the same homoge neous sample un der the prescribed con diti ons.分析過程的精密度是指同一樣品在指定的條件下所做的一系列測試值的相近程度 (離散度)。5.1 Repeatability重復(fù)性Repeatability expresses the precision under the same operating conditions over a short in terval of time.重復(fù)性是指同一個實驗室相同

14、操作條件下短期內(nèi)測定實驗結(jié)果的精密度。5.2 In termediate precisi on中間精密度In termediate precisi on expresses withi n-laboratories variati ons: differe nt days, differe nt an alysts, differe nt equipme nt, etc.中間精密度是指在同一實驗室內(nèi)改變其他條件，包括不同的日期，不同的人，不同 的儀器等的精密度。五、 Project方案內(nèi)容1Accepta nee criteria可接受標(biāo)準(zhǔn)Items確認(rèn)參數(shù)Accepta nee Criter

15、ia可接受標(biāo)準(zhǔn)System suitability 系統(tǒng)適用性Six replicate injections of Standard solution, RSD <3.0%.重復(fù)進(jìn)樣標(biāo)準(zhǔn)溶液6次，RSD <3.0%Resolution > 1.25 (between cytarabine and uridine)阿糖胞苷與尿苷分離度1.25The S/N ratio of the Cytarabine peak and the noise near theCytarabine in the System sensitivity solution is not less th

16、an 10. 系統(tǒng)靈敏度溶液中阿糖胞苷峰與其附近噪音的信噪比不小于10。Specificity專屬性Dilue nt can not have in terferi ng peak that can affect sample measured.空白不含干擾樣品測定的峰。Resolution > 1.25 (between cytarabine and uridine)阿糖胞苷與尿苷分離度1.25Resolution > 1.5 (between uracil and cytarabine, uridine and uracil arab ino side)尿嘧啶與阿糖胞苷、尿苷與

17、阿糖尿苷分離度均1.5Limit of Detect ion (LOD) 檢測限Cytarab ine 阿糖胞苷S/N >3, LOD of each specimenW 0.015% (concentration :0.75 口 g/mL)S/N > 3，每個物質(zhì)的 LOD W 0.015%(濃度:0.75 口 g/mL)Uracil尿嘧啶Uridi ne尿苷UracilArab ino side阿糖尿苷Limit of Quan titati on (LOQ) 定量限Cytarab ine 阿糖胞苷S/N> 10, LOQ of each specimenW 0.05%

18、(concentration :2.5 g/mL)S/N > 10,每個物質(zhì)的 LOQ W0.05% (濃度:2.5 口 g/n)Uracil尿嘧啶Uridi ne尿苷Uracilarab ino side阿糖尿苷Precisio n精密度Repeatability 重復(fù)性RSDW 20% (for 6 results of in dividual impurity which is above 0.03% and the total impurities)RSDW 20%(對于6次試驗測得的大于0.03%的每個雜質(zhì)和總雜 質(zhì))In termediate precisi on 中間精密度

19、RSDW 20% (for 6 results of in dividual impurity which is above 0.03% and the total impurities)RSDW 20%(對6次試驗測得的大于 0.03%每個雜質(zhì)和總雜質(zhì))RSDW 30% (for 12 results of in dividual impurity which is above 0.03% and the total impurities)RSDW 30%(對12次試驗測得的大于 0.03%每個雜質(zhì)和總雜質(zhì))Soluti on stability溶液穩(wěn)定性System suitability

20、 solution: three peaks of Cytarabine, Uridine and Uracil arab ino side exist.系統(tǒng)適用性溶液：阿糖胞苷、尿苷與阿糖尿苷3個峰存在。Standard solution: recovery from aged standard solution isbetween 90.0%110.0% against that from initial results. 標(biāo)準(zhǔn)溶液：與初始結(jié)果比較，放置后的標(biāo)準(zhǔn)溶液回收率在 90.0%110.0% 之間。Test solution: the RSD of % impurity result

21、s which is above 0.03% and the total impurity between aged test solution and the initial results is NMT 30%.供試品溶液：放置后的供試液中與初始結(jié)果中大于0.03%的雜質(zhì)及總雜質(zhì)含量RSD不大于30%。Mobile phase stability 流動相穩(wěn)定性The appearance is clear, without apparent change.外觀應(yīng)澄清無明顯變化。Six replicate injections of standard solution, RSD < 3

22、.0%.重復(fù)進(jìn)標(biāo)準(zhǔn)溶液 6次，RSDW 3.0%Resolution > 1.25 (between Cytarabine and Uridine) 阿糖胞苷與尿苷分離度1.25The S/N ratio of the Cytarab ine peak and the no ise n ear the Cytarabine in the System sensitivity solution is not less than 10. 系統(tǒng)靈敏度溶液中阿糖胞苷峰與其附近噪音的信噪比不小于10。2Reage nts, sample, refere nee sta ndards, and in

23、 strume nts試劑、樣品、標(biāo)準(zhǔn)品、儀器2.1 Reage nts試劑Mono basic sodium phosphate (HPLC) 磷酸二氫鈉(HPLC)Dibasic sodium phosphate (HPLC) 磷酸氫二鈉(HPLC)Sodium hydroxide (AR)氫氧化鈉(AR)Phosphoric acid (AR) 磷酸(AR)Metha nol (HPLC)甲醇(HPLC)Uridi ne (AR)尿苷(AR)Uracil (AR)尿嘧啶(AR)Water(purified, fresh daily)水(純化水，每天新制)2.2 Sample樣品Cytar

24、abine (Zhejiang Hisun Pharmaceutical Co. Ltd, 1 batch)阿糖胞苷(浙江海正藥業(yè)股份有限公司，1批)2.3 Refere nee sta ndards標(biāo)準(zhǔn)品USP Cytarabine RS(current lot) 阿糖胞苷USP標(biāo)準(zhǔn)品(現(xiàn)行批)USP Uracil arabi noside RS(curre nt lot)阿糖尿苷USP標(biāo)準(zhǔn)品(現(xiàn)行批)2.4 In strume nts儀器HPLC in strume nt液相色譜儀Electr onic bala nee電子天平pH meterpH計Waters e2695XS205 Du

25、al rangMetrohm 8273Test method測試方法USP 34 Cytarabine( in the appendix)3.1 Chromatographic con diti on色譜條件Column:Supelcosil LC-18-DB, 250 4.6mm, 5(im色譜柱：See gradie nt見梯度1.0 mL/min254 nmMobile phase:流動相：Flow Rate:流速：Detecti on wavele ngth:檢測波長：Column temperature: 20 C 柱溫：Injection volume:20 口1進(jìn)樣體積：Phos

26、phate buffer: 磷酸鹽緩沖液：Prepare a soluti on containing 0.01M mono basic sodium phosphate and 0.01M dibasic sodium phosphate in a suitable container. Adjust with 0.1M sodium hydroxide or 0.1M phosphoric acid to a pH of 7.0.配制濃度為0.01M的磷酸二氫鈉和0.01M的磷酸氫二鈉溶液，用0.1M氫氧化鈉或0.1M 磷酸調(diào)pH至7.0。Solution A溶液APrepare a fi

27、ltered and degassed mixture of Phosphate buffer and methanol (49:1).Make adjustments if necessary. Prepare this solution fresh daily.磷酸緩沖鹽溶液：甲醇(49:1),過濾脫氣即得。必要時調(diào)整比例，每天新配。Solution B:溶液BPrepare a filtered and degassed mixture of Phosphate buffer and methanol (7:3).Make adjustments if necessary. Prepar

28、e this solution fresh daily.磷酸緩沖鹽溶液：甲醇(7:3)，過濾脫氣即得。必要時調(diào)整比例，每天新配。Gradient:梯度Time (mi nute)時間(分鐘)Solution A溶液ASolution B溶液B01000101000200100250100301000501000System suitability soluti on: 系統(tǒng)適用性溶液：Weigh accurately about 20 mg of Uridine, transfer into a 200-mL volumetric flask, dissolve and dilute with

29、 water to volume, mix well to obtain Uridine Stock Solution, the solution has a concentration of about 0.1mg/mL.精密稱取約20mg尿苷至200mL容量瓶中，用水溶解后稀釋至刻度，混勻，作為尿苷 儲備液，溶液濃度大約是 0.1mg/mL。Weigh accurately about 20 mg of USP Uracil arabi no side RS , tra nsfer into a 200-mL volumetric flask, dissolve and dilute wi

30、th water to volume, mix well to obta in Uracil arabinoside Stock Solution , the solution has a concentration of about 0.1mg/mL.精密稱取約20mg阿糖尿苷USP標(biāo)準(zhǔn)品至200mL容量瓶中，用水溶解后稀釋至刻度，混勻，作為阿糖尿苷儲備液，溶液濃度大約是0.1mg/mLWeigh accurately about 25 mg of USP Cytarab ine RS, tran sfer into a 5-mL volumetric flask, tran sfer 1.

31、0 mL of Uridi ne Stock Solution and 1.0 mL of Uracil Arabi no side Stock Soluti on in to the volumetric flask, dissolve and dilute to volume with water, mix well to obtai n System suitability soluti on, the soluti on con tai n about 0.02, 0.02, 5.0 mg/mL, respectively.精密稱定25mg阿糖胞苷USP標(biāo)準(zhǔn)品至5mL容量瓶中，分別加入

32、1.0mL尿苷儲備液 和1.0mL阿糖尿苷儲備液，用水溶解后稀釋至刻度，混勻，即得系統(tǒng)適用性溶液， 濃度分別為 0.02、0.02、5.0mg/ml。Stan dard soluti on:標(biāo)準(zhǔn)液：Weigh accurately about 20 mg of USP Cytarabi ne RS, tran sfer into a 200-mL volumetric flask, dissolve and dilute with water to volume, mix well. Tran sfer 2.0mL of obta ined soluti on to a 50-mL volum

33、etric flask, dilute with water to volume and mix well, the solution has concentration of about 4 g /mL 。 卩精密稱取約20mgUSP的阿糖胞苷標(biāo)準(zhǔn)品至200mL容量瓶中，用水溶解后稀釋至刻度， 混勻。精密移取該溶液2.0mL置50mL量瓶中，加水稀釋至刻度，搖勻，即得，濃度約為4由/mL。Test soluti on:供試液：Tran sfer about 25mg of cytarab ine, accurately weighed, to a 5.0-mL volumetric flas

34、k, dissolve in and dilute with water to volume, and mix, the soluti on has a concen trati on of about 5mg/mL. NOTE-prepare this soluti on fresh daily.精密稱取阿糖胞苷約25mg,置5mL容量瓶中，用水溶解并稀釋至刻度，混勻，濃度 約為5mg/mL。(注：此溶液每天新配。)3.2 Procedure實驗過程Inject equal volumes (about 20口 LSystem suitability solution, the Standa

35、rd solutionand the Test soluti on into the chromatograph, record the chromatograms, and measure the peak resp on ses. Calculate the perce ntage of uracil arab ino side in the porti on of Cytarab ine take n by the formula:將相同體積(約20 口 L的系統(tǒng)適用性溶液、標(biāo)準(zhǔn)液和供試液分別注入色譜儀中，記 錄色譜圖，計算峰面積。用下式計算阿糖胞苷中阿糖尿苷的含量：P(Cs / Ci

36、)(ri/1.34rs)in which,其中，P is the conten t of USP Cytarabi ne RS in the Sta ndard solutio n;P為標(biāo)準(zhǔn)液中阿糖胞苷 USP標(biāo)準(zhǔn)品的含量Cs is the concen trati onof USP Cytarabi ne RS in the Stan dard soluti on, mg / mL;Cs為標(biāo)準(zhǔn)液中阿糖胞苷 USP標(biāo)準(zhǔn)品的濃度，mg/mL ;Ci is the concen trati on of Cytarab ine in the Test soluti on, mg / mL;Ci為(阿

37、糖胞苷)樣品濃度， mg/mL ;1.34 is the relative resp onse factor for uracil arab ino side;1.34為阿糖尿苷相對響應(yīng)因子；ri is the peak resp onse of uracil arab ino side in the Test soluti on;ri為供試液中阿糖尿苷的峰響應(yīng)值；rs is the peak resp onse of USP Cytarab ine RS in theSta ndard soluti on:rs為標(biāo)準(zhǔn)液中阿糖胞苷 USP標(biāo)準(zhǔn)品的峰響應(yīng)值。Not more than 0.20%

38、 is found.阿糖尿苷含量不得過 0.20%。Calculate the perce ntage of all other impurities in the porti on of Cytarab ine take n by the formula:其它雜質(zhì)含量按下式計算：P(Cs / Ci )(ri/Frs)in which,式中P is the conten t of USP Cytarabi ne RS in the Sta ndard solutio n;P為標(biāo)準(zhǔn)液中阿糖胞苷 USP標(biāo)準(zhǔn)品的含量Cs is the concen trati onof USP Cytarabi n

39、e RS in the Stan dard soluti on, mg / mL;Cs為標(biāo)準(zhǔn)液中阿糖胞苷 USP標(biāo)準(zhǔn)品的濃度，mg/mL ；Ci is the concen trati on of Cytarab ine in the Test soluti on, mg/mL;Ci為(阿糖胞苷)樣品濃度， mg/mL ；ri is the peak resp onse of each impurity in the Test soluti on;ri為供試液中各單個雜質(zhì)的峰面積；rs is the peak resp onse of USP Cytarab ine RS in theSta

40、ndard soluti on;rs為標(biāo)準(zhǔn)液中阿糖胞苷 USP標(biāo)準(zhǔn)品的峰面積；F is the relative resp onse factor, equals 2.5 for the Uracil peak, with a relative rete nti ontime time of 0.55, 1.5 for peaks with relative rete nti on times of 0.38, 0.43 and 1.14, and 1.0 for all other peaks.F為相對響應(yīng)因子，尿嘧啶相對保留時間為0.55,相對響應(yīng)因子為2.5，相對保留時間為0.38、0.

41、43、1.14的雜質(zhì)，相對響應(yīng)因子為 1.5，其它峰為1.0。Not more tha n 0.10% of any in dividual impurity is found, and not more tha n 0.30% of total impurities is found (in cludi ng uracil arab ino side).單個雜質(zhì)不得過0.10%，總雜質(zhì)不得過0.30% (包括阿糖尿苷)。Verificati on確認(rèn)4.1System suitability系統(tǒng)適用性Preparati on of soluti ons:溶液制備：Impurity stock

42、 solution :雜質(zhì)儲備液：Weigh accurately about 20 mg of Uridi ne, tran sfer into a 200-mL volumetric flask, dissolve and dilute with water to volume, mix well to obtain Uridine Stock Solution , the soluti on has a concen trati on of about 0.1mg/mL.精密稱取約20mg尿苷至200mL容量瓶中，用水溶解后稀釋至刻度，混勻，作為尿苷 儲備液，濃度大約0.1mg/mL。W

43、eigh accurately about 20 mg of USP Uracil Arabi no side RS , tran sfer into a 200-mL volumetric flask, dissolve and dilute with water to volume, mix well to obta in Uracil Arab ino side Stock Soluti on, the soluti on has a concen trati on of about 0.1mg/mL.精密稱取約20mg阿糖尿苷USP標(biāo)準(zhǔn)品至200mL容量瓶中，用水溶解后稀釋至刻度， 混

44、勻，作為阿糖尿苷儲備液，濃度大約0.1mg/mL。System suitability soluti on:系統(tǒng)適用性測試液：Weigh accurately about 25 mg of USP Cytarab ine RS, tran sfer into a 5-mL volumetric flask, transfer accurately 1.0 mL of Uridine Stock Solution and 1.0 mL of Uracil Arab ino side Stock Soluti on into the volumetric flask, dissolve and d

45、ilute to volume with water, mix well. (0.02mg/mL for Uridi ne, 0.02 mg/mL for Uracil Arab ino side, 5.0 mg/mL for Cytarab in e).精密稱定25mg阿糖胞苷USP標(biāo)準(zhǔn)品至5mL容量瓶中，分別加入1.0mL尿苷儲備液和 1.0mL阿糖尿苷儲備液，用水溶解后稀釋至刻度，混勻(濃度分別為尿苷0.02mg/mL、阿糖尿苷0.02 mg/mL、阿糖胞苷5.0mg/mL )。Stan dard soluti on:標(biāo)準(zhǔn)液：Weigh accurately about 20 mg of

46、 USP Cytarabi ne RS, tran sfer into a 200-mL volumetric flask, dissolve and dilute to volume with water, mix well. Transfer 2.0mL of obtained solution into a 50-mL volumetric flask, dilute with water to volume and mix, the solution has a concentration of about 4口 g/mL.精密稱定約20mg阿糖胞苷USP標(biāo)準(zhǔn)品至200mL容量瓶中，用

47、水溶解并稀釋至刻度。 移取2.0mL至50mL容量瓶中，用水稀釋至刻度并混勻，濃度大約4 口 g/mL。System sen sitivity soluti on:系統(tǒng)靈敏度溶液：Weigh accurately about 20 mg of USP Cytarabi ne RS, tran sfer into a 200-mL volumetric flask, dissolve and dilute to volume with water, mix well. Transfer 2.0 mL of obtained solution into a 100-mL volumetric fl

48、ask, dilute with water to volume and mix, the solution has a concentration of about 2口 g/mL.精密稱定約20mg阿糖胞苷USP標(biāo)準(zhǔn)品至200mL容量瓶中，用水溶解并稀釋至刻度。 移取2.0mL至100mL容量瓶中，用水稀釋至刻度并混勻，濃度大約2 口 g/mL。Procedure實驗過程Separately inject the System suitability solution , the Standard solution and the System sen sitivity soluti on

49、 into the chromatograph, record the chromatograms. Calculate the relative sta ndard deviati on (RSD) of the Stan dard soluti on, the resoluti on of the System suitability solution and S/N of the System sensitivity solution.分別將系統(tǒng)適用性測試液、標(biāo)準(zhǔn)液和系統(tǒng)靈敏度溶液注入色譜，記錄色譜圖。計算 標(biāo)準(zhǔn)溶液的相對標(biāo)準(zhǔn)偏差(RSD),系統(tǒng)適用性溶液的分離度和系統(tǒng)靈敏度溶液的信噪

50、比。Injectio nsS.N.Descripti onNo. of injectio n(s)1Bla nk(water)12System sen sitivity soluti on13System suitability soluti on14Bla nk(water)3 at in itial gradie nt for 12m in*5Stan dard soluti on6To elim in ate carry over of the in jectorAccepta nee criteria可接受標(biāo)準(zhǔn)The S/N ratio of the Cytarabine peak a

51、nd the noise near the Cytarabine in the System sensitivity solution is not less than 10.系統(tǒng)靈敏度溶液中阿糖胞苷峰與其附近噪音的信噪比不小于10。Six replicate injections of Standard solution, RSD <3.0%.標(biāo)準(zhǔn)溶液重復(fù)進(jìn)6次，RSD <3.0%。Resolution between Cytarabine and Uridine in the System suitability solution is > 1.25. 系統(tǒng)適用性測試液中

52、阿糖胞苷與尿苷分離度1.25。4.2 Specificity專屬性4.2.1 Preparati on of soluti ons:溶液制備Impurity stock solution :雜質(zhì)儲備液：Weigh accurately about 20 mg of Uridine, transfer into a 200-mL volumetric flask, dissolve and dilute with water to volume, mix well to obtain Uridine stock solution , the solution has a concentratio

53、n of about 0.1mg/mL.精密稱取約20mg尿苷至200mL容量瓶中，用水溶解后稀釋至刻度，混勻，作為尿苷 儲備液，濃度大約0.1mg/mL。Weigh accurately about 20 mg of USP Uracil Arabi no side RS , tran sfer into a 200-mL volumetric flask, dissolve and dilute with water to volume, mix well to obta in Uracil Arabinoside stock solution, the solution has a co

54、ncentration of about 0.1mg/mL.精密稱取約20mg阿糖尿苷USP標(biāo)準(zhǔn)品至200mL容量瓶中，用水溶解后稀釋至刻度， 混勻，作為阿糖尿苷儲備液，濃度大約0.1mg/mL。Weigh accurately about 20 mg of Uracil , tran sfer into a 200-mL volumetric flask, dissolve and dilute with water to volume, mix well to obtain Uracil stock solution , the solution has a concen trati on

55、 of about 0.1mg/mL.精密稱取約20mg尿嘧啶至200mL容量瓶中，用水溶解后稀釋至刻度，混勻，作為尿 嘧啶儲備液，濃度大約 0.1mg/mL。Selectivity solution :選擇性溶液Weigh accurately about 25 mg of USP Cytarabine RS into a 5-mL volumetric flask, transfer 1.0 mL of Uridine stock solution , 1.0 mL of Uracil stock solution and 1.0 mL of Uracil Arabinoside stoc

56、k solution into the volumetric flask. dissolve and dilute with water to volume, mix well. (0.02mg/mL for Uridine, 0.02mg/mL for Uracil, 0.02 mg/mL forUracil Arab ino side, 5.0 mg/mL for Cytarab ine)精密稱定約25mg阿糖胞苷USP標(biāo)準(zhǔn)品至5mL容量瓶中，分別加入1.0mL尿苷儲備 液，1.0mL尿嘧啶儲備液和1.0mL阿糖尿苷儲備液，用水溶解后稀釋至刻度，混勻(濃度分別為尿苷 0.02mg/mL、尿嘧啶0.02 mg/mL、阿糖尿苷0.02 mg/mL、阿糖胞 苷 5.0mg/mL )。Un spiked test soluti on:供試液Weigh accurately about 25mg of Cytarabine sample into a 5-mL volumetric flask ，dissolve and dilute to volume with water, the solution has a concentration of about 5.0mg/mL.精密稱定約25mg阿糖胞苷樣品至5mL容量瓶中，用水溶解后稀釋至刻度，