á233? ELEMENTAL IMPURITIES—PROCEDURES
INTRODUCTION
This chapter describes two analytical procedures (Procedures 1 and 2) for the evaluation of the levels of the elemental impuri-ties. The chapter also describes criteria for acceptable alternative procedures. By means of validation studies, analysts will confirm that the analytical procedures described herein are suitable for use on specified material.
Use of Alternative Procedures
The chapter also describes criteria for acceptable alternative procedures. Alternative procedures that meet the validation re-quirements herein may be used in accordance with General Notices and Requirements 6.30, Alternative and Harmonized Meth-ods and Procedures . Information on the Requirements for Alternate Procedure Validation is provided later in this chapter.Speciation
The determination of the oxidation state, organic complex, or combination is termed speciation . Analytical procedures for spe-ciation are not included in this chapter, but examples may be found elsewhere in USP–NF and in the literature.
PROCEDURES
? C OMPENDIAL P ROCEDURES 1 AND 2
System standardization and suitability evaluation using applicable reference materials should be performed on the day of analysis.
Procedure and detection technique:Procedure 1 can be used for elemental impurities generally amenable to detection by
inductively coupled plasma–atomic (optical) emission spectroscopy (ICP–AES or ICP–OES). Procedure 2 can be used for ele-mental impurities generally amenable to detection by ICP–MS. Before initial use, the analyst should verify that the proce-
dure is appropriate for the instrument and sample used (procedural verification) by meeting the alternative procedure vali-dation requirements below.
Sample preparation:Forms of sample preparation include Neat , Direct aqueous solution , Direct organic solution , and Indi-
rect solution . The selection of the appropriate sample preparation depends on the material under test and is the responsibil-ity of the analyst. When a sample preparation is not indicated in the monograph, an analyst may use any of the following
appropriately validated preparation procedures. In cases where spiking of a material under test is necessary to provide an acceptable signal intensity, the blank should be spiked with the same Target elements , and where possible, using the same spiking solution. Standard solutions may contain multiple Target elements . [N OTE —All liquid samples should be weighed.]Neat:Used for liquids or alternative procedures that allow the examination of unsolvated samples.
Direct aqueous solution:Used when the sample is soluble in an aqueous solvent.
Direct organic solution:Used where the sample is soluble in an organic solvent.
Indirect solution:Used when a material is not directly soluble in aqueous or organic solvents. Total metal extraction is the preferred sample preparation approach to obtain an Indirect solution . Digest the sample using the Closed vessel diges-tion procedure provided below or one similar to it. The sample preparation scheme should yield sufficient sample to allow quantification of each element at the limit specified in the corresponding monograph or chapter.
Closed vessel digestion:This sample preparation procedure is designed for samples that must be digested in a Concen-trated acid using a closed vessel digestion apparatus. Closed vessel digestion minimizes the loss of volatile impurities. The choice of a Concentrated acid depends on the sample matrix. The use of any of the Concentrated acids may be appropri-ate, but each introduces inherent safety risks. Therefore, appropriate safety precautions should be used at all times.
[N OTE —Weights and volumes provided may be adjusted to meet the requirements of the digestion apparatus used.]
An example procedure that has been shown to have broad applicability is the following. Dehydrate and predigest 0.5 g of primary sample in 5 mL of freshly prepared Concentrated acid . Allow to sit loosely covered for 30 min in a fume hood.Add an additional 10 mL of Concentrated acid , and digest, using a closed vessel technique, until digestion or extraction is complete. Repeat, if necessary, by adding an additional 5 mL of Concentrated acid . [N OTE —Where closed vessel digestion is necessary, follow the manufacturer’s recommended procedures to ensure safe use.]
Alternatively, leachate extraction may be appropriate with justification following scientifically validated metal disposition studies, which may include animal studies, speciation, or other means of studying disposition of the specific metal in the drug product.
Reagents:All reagents used for the preparation of sample and standard solutions should be free of elemental impurities,in accordance with Plasma Spectrochemistry á730?.
? P ROCEDURE 1: ICP–OES
Standardization solution 1: 1.5J of the Target element(s) in a Matched matrix
Standardization solution 2:0.5J of the Target element(s) in a Matched matrix
Sample stock solution:Proceed as directed in Sample preparation above. Allow the sample to cool, if necessary. For mer-cury determination, add an appropriate stabilizer.
Sample solution:Dilute the Sample stock solution with an appropriate solvent to obtain a final concentration of the Target elements at NMT 1.5J .
Blank:
Matched matrix
298 á233? Elemental Impurities—Procedures / Chemical Tests USP 40
Elemental spectrometric system
(See Plasma Spectrochemistry á730?.)
Mode:ICP
Detector:Optical detection system
Rinse:Diluent used
Standardization:Standardization solution 1, Standardization solution 2, and Blank
System suitability
Sample:Standardization solution 1
Suitability requirements
Drift:Compare results obtained from Standardization solution 1 before and after the analysis of the Sample solution.
Suitability criteria:NMT 20% for each Target element. [N OTE—If samples are high in mineral content, rinse system well before introducing the Sample in order to minimize carryover.]
Analysis:Analyze according to the manufacturer's suggestions for program and wavelength. Calculate and report results on the basis of the original sample size. [N OTE—Appropriate measures must be taken to correct for matrix-induced inter-ferences (e.g., wavelength overlaps).]
? P ROCEDURE2: ICP–MS
Standardization solution 1: 1.5J of the Target element(s) in a Matched matrix
Standardization solution 2:0.5J of the Target element(s) in a Matched matrix
Sample stock solution:Proceed as directed for Sample preparation above. Allow the sample to cool, if necessary. For mercury determination, add an appropriate stabilizer.
Sample solution:Dilute the Sample stock solution with an appropriate solvent to obtain a final concentration of the Target elements at NMT 1.5J.
Blank:Matched matrix
Elemental spectrometric system
(See Plasma Spectrochemistry á730?.)
Mode:ICP. [N OTE—An instrument with a cooled spray chamber is recommended. (A collision cell or reaction cell may also be beneficial.)]
Detector:Mass spectrometer
Rinse:Diluent used
Standardization:Standardization solution 1, Standardization solution 2, and Blank
System suitability
Sample:Standardization solution 1
Suitability requirements
Drift:Compare results obtained from Standardization solution 1 before and after the analysis of the Sample solution.
Suitability criteria:Drift NMT 20% for each Target element. [N OTE—If samples are high in mineral content, rinse sys-tem well before introducing the Sample in order to minimize carryover.]
Analysis:Analyze according to the manufacturer's suggestions for program and m/z. Calculate and report results based on the original sample size. [N OTE—Appropriate measures must be taken to correct for matrix-induced interferences (e.g., argon chloride interference with arsenic determinations).]
REQUIREMENTS FOR ALTERNATE PROCEDURE VALIDATION
If the specified compendial procedures do not meet the needs of a specific application, an alternative procedure may be devel-oped (see General Notices and Requirements 6.30, Alternative and Harmonized Methods and Procedures). Alternative proce-dures must be validated and shown to be acceptable, in accordance with the validation requirements for alternative proce-dures as described below. The level of validation necessary to ensure that an alternative procedure is acceptable depends on whether a limit test or a quantitative determination is specified in the monograph. The requirements for the validation of an elemental impurities procedure for each type of determination are described below. Any alternative procedure that has been validated and meets the acceptance criteria that follow is considered to be suitable for use.
LIMIT PROCEDURES
The following section defines the validation parameters for the acceptability of alternative limit procedures. Meeting these re-quirements must be demonstrated experimentally using an appropriate system suitability procedure and reference material. The suitability of the method must be determined by conducting studies with the material or mixture under test supplemen-ted with known concentrations of each Target element of interest at the appropriate acceptance limit concentration. The ma-terial or mixture under test must be spiked before any sample preparation steps are performed.
? D ETECTABILITY
Standard solution: A preparation of reference materials for the Target element(s) at the Target concentration
Spiked sample solution 1:Prepare a solution of sample under test, spiked with appropriate reference materials for the Target elements at the Target concentration, solubilized or digested as described in Sample preparation.
Spiked sample solution 2:Prepare a solution of the sample under test, spiked with appropriate reference materials at 80% of the Target concentration for the Target elements, solubilized or digested as described in Sample preparation.
Unspiked sample solution: A sample of material under test, solubilized or digested in the same manner as the Sample
solutions
USP 40Chemical Tests / á233? Elemental Impurities—Procedures 299
Acceptance criteria
Non-instrumental procedures:Spiked sample solution 1 provides a signal or intensity equivalent to or greater than that of the Standard solution . Spiked sample solution 2 must provide a signal or intensity less than that of Spiked sample solu-tion 1. [N OTE —The signal from each Spiked sample solution is NLT the Unspiked sample solution determination.]
Instrumental procedures:The average value of the three replicate measurements of Spiked sample solution 1 is within ±15% of the average value obtained for the replicate measurements of the Standard solution . The average value of the replicate measurements of Spiked sample solution 2 must provide a signal intensity or value less than that of the Standard solution . [N OTE —Correct the values obtained for each of the spiked solutions using the Unspiked sample solution .]
? P RECISION FOR I NSTRUMENTAL M ETHODS (R EPEATABILITY )
[N OTE —Non-instrumental precision is demonstrated by meeting the Detectability requirement above.]
Sample solutions:Six independent samples of the material under test, spiked with appropriate reference materials for the Target elements at the Target concentration
Acceptance criteria
Relative standard deviation:NMT 20% for each Target element
? S PECIFICITY
The procedure must be able to unequivocally assess (see Validation of Compendial Procedures á1225?) each Target element in the presence of components that may be expected to be present, including other Target elements , and matrix compo-nents.
QUANTITATIVE PROCEDURES
The following section defines the validation parameters for the acceptability of alternative quantitative procedures. Meeting
these requirements must be demonstrated experimentally, using an appropriate system suitability procedure and reference
materials. Meeting these requirements demonstrates that the procedure is equivalent to the compendial procedure for the
purpose of quantifying the Target elements .
? A CCURACY
Standard solutions:Prepare solutions containing the Target elements at concentrations ranging from 50% to 150% of J ,
using appropriate reference materials.
Test samples:Prepare samples of the material under test spiked with appropriate reference materials before any sample preparation steps (digestion or solubilization) at concentrations ranging from 50% to 150% of J for each Target element .
Acceptance criteria
Spike recovery:70%–150% for the mean of three replicate preparations at each concentration
? P RECISION
Repeatability
Test samples:Six independent samples of material under test (taken from the same lot) spiked with appropriate refer-ence materials for the Target element(s) at the indicated level
Acceptance criteria
Relative standard deviation:NMT 20% (N = 6) for each Target element
Intermediate precision (ruggedness)
Perform the Repeatability analysis again either on a different day, with a different instrumentation, with a different analyst,or a combination thereof. Combine the results of this analysis with the Repeatability analysis so the total number of anal-yses is 12.
Acceptance criteria
Relative standard deviation:NMT 25% (N = 12) for each Target element
? S PECIFICITY
The procedure must be able to unequivocally assess (see á1225?) each Target element in the presence of components that may be expected to be present, including other Target elements , and matrix components.
? L IMIT OF Q UANTITATION , R ANGE , AND L INEARITY
Demonstrated by meeting the Accuracy requirement.
GLOSSARY
Concentrated acid:Concentrated ultra-pure nitric, sulfuric, hydrochloric, or hydrofluoric acids or Aqua regia
Aqua regia:Aqua regia is a mixture of concentrated hydrochloric and nitric acids, typically at ratios of 3:1 or 4:1, respective-ly.
Matched matrix:Solutions having the same solvent composition as the Sample solution . In the case of an aqueous solution,Matched matrix would indicate that the same acids, acid concentrations, and mercury stabilizer are used in both prepara-tions.
Target elements:Elements with the potential of being present in the material under test. Include arsenic (As), cadmium (Cd), lead (Pb), and mercury (Hg) in the target element evaluation when testing is done to demonstrate compliance. Target elements should also include any elements that may be added through material processing or storage.
Target limit or Target concentration:The acceptance value for the elemental impurity being evaluated. Exceeding the Tar-get limit indicates that a material under test exceeds the acceptable value. The determination of compliance is addressed in other chapters. [N OTE —When applying this chapter to Elemental Impurities—Limits á232? and
Elemental Contaminants in Diet-
300 á233? Elemental Impurities—Procedures / Chemical Tests USP 40
ary Supplements á2232?, Target limits can be approximated by dividing the Daily Dose PDEs by the maximum daily dose for the Drug Product Analysis Option in á232? or the Daily Serving PDE divided by the maximum daily serving size in á2232?.]
J:The concentration (w/w) of the element(s) of interest at the Target limit, appropriately diluted to the working range of the instrument. For example, if the target elements are lead and arsenic for an analysis of an oral solid drug product with a daily dose of 10g/day using inductively coupled plasma–mass spectrometry (ICP–MS), the target limit for these elements would be 0.5 m g/g and 1.5 m g/g (see Table 2 in á232?). However, in this case, the linear dynamic range of the ICP–MS is known to extend from 0.01 ng/mL to 0.1 m g/mL for these elements. Therefore, a dilution factor of at least 1:100 is required to ensure that the analysis occurs in the linear dynamic range of the instrument. J would thus equal 5 ng and 15 ng/mL for lead and arsenic, respectively, when the dilution factor is added.
Appropriate reference materials:Where Appropriate reference materials are specified in the chapter, certified reference ma-terials (CRM) from a national metrology institute (NMI), or reference materials that are traceable to the CRM of an NMI should be used. An example of an NMI in the United States is the National Institute of Standards and Technology.
á241? IRON
This limit test is provided to demonstrate that the content of iron, in either the ferric or the ferrous form, does not exceed the limit for iron specified in the individual monograph. The determination is made by concomitant visual comparison with a control prepared from a standard iron solution.
SPECIAL REAGENTS
Standard Iron Solution
Dissolve 863.4 mg of ferric ammonium sulfate [FeNH
4
(SO
4
)
2
·12H
2
O] in water, add 10 mL of 2N sulfuric acid, and dilute with water to 100.0 mL. Pipet 10 mL of this solution into a 1000-mL volumetric flask, add 10 mL of 2N sulfuric acid, dilute with water to volume, and mix. This solution contains the equivalent of 0.01 mg (10 m g) of iron per mL.
Ammonium Thiocyanate Solution
Dissolve 30g of ammonium thiocyanate in water to make 100 mL.
STANDARD PREPARATION
Into a 50-mL color-comparison tube pipet 1 mL of Standard Iron Solution (10 m g of Fe), dilute with water to 45 mL, add 2 mL of hydrochloric acid, and mix.
TEST PREPARATION
Into a 50-mL color comparison tube place the solution prepared for the test as directed in the individual monograph and if necessary dilute with water to 45 mL; or, dissolve in water, and dilute with water to 45 mL the quantity, in g, of the substance to be tested, as calculated by the formula:
1.0/(1000L)
in which L is the Iron limit in percentage. Add 2 mL of hydrochloric acid, and mix.
PROCEDURE
To each of the tubes containing the Standard Preparation and the Test Preparation add 50 mg of ammonium peroxydisulfate crystals and 3 mL of Ammonium Thiocyanate Solution, and mix: the color of the solution from the Test Preparation is not darker than that of the solution from the
Standard Preparation.
USP 40Chemical Tests / á241? Iron 301
0821重金属检查法 本法所指的重金属系指在实验条件下能与硫代乙酰胺或硫化钠作用显色的金属杂质。 标准铅溶液的制备称取硝酸铅0.1599g，置1000ml量瓶中，加硝酸5ml 与水50ml溶解后，用水稀释至刻度，摇匀，作为贮备液。 精密量取贮备液10ml，置100ml量瓶中，加水稀释至刻度，摇匀，即得(每1ml相当于10μg的Pb)。本液仅供当日使用。 配制与贮存用的玻璃容器均不得含铅。 第一法 除另有规定外，取25ml纳氏比色管三支，甲管中加标准铅溶液一定量与醋酸盐缓冲液(pH3.5)2ml后，加水或各品种项下规定的溶剂稀释成25ml，乙管中加入按各品种项下规定的方法制成的供试品溶液25ml；丙管中加入与乙管相同重量的供试品，加配制供试品溶液的溶剂适量使溶解，再加与甲管相同量的标准铅溶液与醋酸盐缓冲液（pH3.5）2ml后，用溶剂稀释成25ml；若供试液带颜色，可在甲管中滴加少量的稀焦糖溶液或其他无干扰的有色溶液，使之与乙管、丙管一致；再在甲、乙、丙三管中分别加硫代乙酰胺试液各2ml，摇匀，放置2分钟，同置白纸上，自上向下透视，当丙管中显出的颜色不浅于甲管时，乙管中显示的颜色与甲管比较，不得更深。如丙管中显示出的颜色浅于甲管，应取样按第二法重新检查。 如在甲管中滴加稀焦糖溶液或其他无干扰的有色溶液，仍不能使颜色一致时，应取样按第二法检查。 供试品如含高铁盐影响重金属检查时，可在甲、乙、丙三管中分别加入相同量的维生素C0.5～1.0g，再照上述方法检查。 配制供试品溶液时，如使用的盐酸超过1 ml，氨试液超过2ml，或加入其他试剂进行处理者，除另有规定外，甲管溶液应取同样同量的试剂置瓷皿中蒸干后，加醋酸盐缓冲液（pH3.5）2ml与水15ml，微热溶解后，移置纳氏比色管中，加标准铅溶液一定量，再用水或各品种项下规定的溶剂稀释成25ml。 第二法 除另有规定外，当需改用第二法检查时，取各品种项下定量的供试品，按炽灼残渣检查法（通则0841）进行炽灼处理，然后取遗留的残渣；或直接取炽灼残渣项下遗留的残渣；如供试品为溶液，则取各品种项下规定量的溶液，蒸
<711> DISSOLUTION 溶出度 (USP39-NF34 Page 540) General chapter Dissolution <711> is being harmonized with the corresponding texts of the European Pharmacopoeia and/or the Japanese Pharmacopoeia. These pharmacopeias have undertaken to not make any unilateral change to this harmonized chapter. 通则<711>溶出度与欧盟药典和日本药典中的相应部分相统一。这三部药典承诺不做单方面的修改。 Portions of the present general chapter text that are national USP text, and therefore not part of the harmonized text, are marked with symbols to specify this fact. 本章中的部分文字为本国USP内容，并没有与其他药典统一。此部分以（）标注。 This test is provided to determine compliance with the dissolution requirements where stated in the individual monograph for dosage forms administered orally. In this general chapter, a dosage unit is defined as 1 tablet or 1 capsule or the amount specified. Of the types of apparatus designs described herein, use the one specified in the individual monograph. Where the label states that an article is enteric coated and a dissolution or disintegration test does not specifically state that it is to be applied to delayed-release articles and is included in the individual monograph, the procedure and interpretation given for Delayed-Release Dosage Forms are applied, unless otherwise specified in the individual monograph. 本测试用于检测药品口服制剂的溶出度是否符合各论中的规定。本章中，除另有规定外，单位制剂定义为1片或1粒胶囊。对于本章中所述多种仪器，使用各论中规定的种类。除各论中另有规定外，如果检品是肠溶衣片且各论中的溶出度或崩解时限检查项下没有特别指出适用迟释剂的，使用本章中适用于迟释剂的流程和解释。 FOR DOSAGE FORMS CONTAINING OR COATED WITH GELATIN涂有或包含明胶的剂型 If the dosage form containing gelatin does not meet the criteria in the appropriate Acceptance Table (see Interpretation, Immediate-Release Dosage Forms, Extended-Release Dosage Forms, or Delayed-Release Dosage Forms) because of evidence of the presence of cross-linking, the dissolution procedure should be repeated with the addition of enzymes to the medium, as described below, and the dissolution results should be evaluated starting at the first stage of the appropriate Acceptance Table. It is not necessary to continue testing through the last stage (up to 24 units) when criteria are not met during the first stage testing, and evidence of cross-linking is observed. 如果剂型中含有明胶，其不符合验收表中的标准（见判断，速释制剂，延释制剂，缓释制剂），因为存在明胶交联结合作用，它的溶解过程与外加的媒介酶是重复的，见下面的描述，并且溶解结果可以通过适当的验收表的开始的第一阶段标准进行评估。如果溶出结果不满足第一阶段的测试标准，那么就没有必要继续测试到最后阶段，并且也证明了明胶交联结合作用的存在。
美国药典(USP)规定的色谱柱编号 L1和L8是美国药典(USP)规定的色谱柱编号，其实就是ODS柱和NH2柱。下面是USP规定的编号所对应的色谱柱类型。 L1：十八烷基键合多孔硅胶或无机氧化物微粒固定相，简称ODS柱 L2：30～50m m表面多孔薄壳型键合十八烷基固定相，简称C18柱 L3：多孔硅胶微粒，即一般的硅胶柱 L4：30～50m m表面多孔薄壳型硅胶柱 L5：30～50m m表面多孔薄壳型氧化铝柱 L6：30～50m m实心微球表面包覆磺化碳氟聚合物，强阳离子交换柱 L7：全多孔硅胶微粒键合C8官能团固定相，简称C8柱 L8：全多孔硅胶微粒键合非交联NH2固定相，简称NH2柱 L9：强酸性阳离子交换基团键合全多孔不规则形硅胶固定相，即SCX柱 L10：多孔硅胶微球键合氰基固定相（CN），简称CN柱 L11：键合苯基多孔硅胶微球固定相，简称苯基柱 L12：无孔微球键合季胺功能团的强阴离子交换柱 L13：三乙基硅烷化学键合全多孔硅胶微球固定相（C1），简称C1柱 L14：10m m硅胶化学键合强碱性季铵盐阴离子交换固定相，简称SAX柱 L15：已基硅烷化学键合全多孔硅胶微球固定相，简称C6柱 L16：二甲基硅烷化学键合全多孔硅胶微粒固定相 C2柱 L17：氢型磺化交联苯乙烯-二乙烯基苯共聚物,强阳离子交换柱 L18：3～10m m全多孔硅胶化学键合胺基（NH2）和氰基（CN）柱 L19：钙型磺化交联苯乙烯－二乙烯基苯共聚物，强阳离子交换柱 L20：二羟基丙烷基化学键合多孔硅胶微球固定相（Diol），简称二醇基柱 L21：刚性苯乙烯－二乙烯基苯共聚物微球填料柱
231 重金属检查法 本试验系在规定的试验条件下，金属离子与硫化物离子反应显色，通过与制备的标准铅溶液目视比较测定， 以确证供试品中重金属杂质含量不超过各论项下规定的限度（以供试品中铅的百分比表示，以重量计） 。【见分光 光度法和光散射项下测定法目视比较法 <851> 】【注意：对本试验有反应的典型物质有铅、汞、铋、砷、锑、锡、 镉、银、铜和钼等】 除各论另有规定外，按第一法测定重金属。第一法适用于在规定试验条件下，能产生澄清、无色溶液的物质。 第二法适用于在第一法规定试验条件下不能产生澄清、无色溶液的物质，或者适用于由于性质复杂， 化物离子与金属离子形成沉淀的物质，或者是不易挥发的和易挥发的油类物质。第三法为湿消化法，仅用于第一 法、第二法都不适合的情况。 特殊试剂 硝酸铅贮备液制备：取硝酸铅 159.8mg ， 溶于 100ml 水中，加 1ml 硝酸，用水稀释至 1000ml 。制备和贮存 本溶液的玻璃容器应不含可溶性铅。 标准铅溶液制备：使用当天，取硝酸铅贮备液 10.0ml ， 用水稀释至 100.0ml 。每 1mL 的标准铅溶液含相当 于 10μg 的铅。按每克供试品取 100μL 标准铅溶液制备的对照溶液， 相当于供试品含百万分之一的铅。 方法Ⅰ pH3.5 醋酸盐缓冲液的制备： 取醋酸铵 25.0g 溶于 25mL 水中， 加 6N 盐酸液 38.0mL ， 必要时， 用 6N 氢氧化铵液或 6N 盐 酸液调节 pH 至 3.5 ， 用水稀释至 100mL ， 混匀。 取标准铅溶液 2.0mL （相当于 20 μg 铅）于50mL 比色管中， 加水稀释至 25mL ， 以 pH 计或精密 pH 试纸作为外 指示剂，用 1N 醋酸液或 6N 氢氧化铵液调节 pH 至 3.0~4.0 ， 用水稀释至 40mL ， 混匀。 取各论项下规定的供试品溶液 25mL 于 50mL 比色管中， 或用各论项下规定用 量的酸溶液样品， 用水稀释至 25mL ，供试品以 g 计，按下式计算： 2.0/ （1000L ） 式中 L 是重金属限度（ % ）。以 pH 计或是精密 pH 试纸作为外指示剂，用 1N 醋酸液或 6N 氢氧化铵液调 节 pH 至 3.0~4.0 ， 用水稀释至 40mL ， 混匀。 取供试品溶液制备项下的溶液 25mL 于 50mL 比色管中， 加标准铅溶液 2.0mL ，以 pH 计或是精密 pH 试纸 作为外指示剂，用 1N 醋酸液或 6N 氢氧化 铵液调节 pH 至 3.0~4.0 ， 用水稀释至 40mL ， 混匀。 在上述三试管中，分别加入 pH3.5 的醋酸盐缓冲液 2mL ， 然后再加硫代乙酰 胺－甘油试液 1.2mL ，用水稀 释至 50mL ， 混匀，放置 2 分钟， 在白色平面 自上向下观察：供试品溶液产生的颜色与标准 品溶液产生的颜色相比，不得更 深。对照溶液产生的颜色比标准溶液深或相当。 ［注意： 如果 对照溶液的颜色比 标准溶液浅，用方法 II 代替方法 I 测定供试品 ］。 易干扰硫 标准溶液制备： 供试品溶液制备： 对照溶液制备 检查法：
美国药典USP31-NF26无菌检查法《71》.doc 71 STERILITY TESTS 无菌检查法 此通则的各部分已经与欧洲药典和/或日本药典的对应部分做了协调。不一致的部分用符号（）来标明。 下面这些步骤适用于测定是否某个用于无菌用途的药品是否符合其具体的各论中关于无菌 检查的要求。只要其性质许可，这些药品将使用供试产品无菌检查法项下的膜过滤法来检测。如果膜过滤技术是不适合的，则使用在供试产品无菌检查法项下的培养基直接接种法。除了具有标记为无菌通道的设备之外，所有的设备均须使用培养基直接接种法进行检测。在结果的观测与理解项下包含了复验的规定。 由于无菌检查法是一个非常精确的程序，在此过程中程序的无菌状态必须得到确保以实现对结果的正确理解，因此人员经过适当的培训并取得资质是非常重要的。无菌检查在无菌条件下进行。为了实现这样的条件，试验环境必须调整到适合进行无菌检查的方式。为避免污染而采取的特定预防措施应不会对任何试图在检查中发现的微生物产生影响。通过在工作区域作适当取样并进行适当控制，来定期监测进行此试验的工作条件。 这些药典规定程序自身的设计不能确保一批产品无菌或已经灭菌。这主要是通过灭菌工艺或者无菌操作程序的验证来完成。 当通过适当的药典方法获得了某物品中微生物污染的证据，这样获得的结果是该物品未能达到无菌检验要求的结论性证据，即便使用替代程序得到了不同的结果也无法否定此结果。如要获得关于无菌检验的其他信息，见药品的灭菌和无菌保证<1211> 按照下面描述的方法配制实验用培养基；或者使用脱水培养基，只要根据其制造商或者分销商说明进行恢复之后，其能够符合好氧菌、厌氧菌、霉菌生长促进试验的要求即可。使用经过验证的工艺对培养基进行灭菌操作。 下面的培养基已经被证实适合进行无菌检查。巯基醋酸盐液体培养基主要用于厌氧菌的培养。但其也用于检测好氧菌。大豆酪蛋白消化物培养基适合于培养霉菌和好氧菌。 Fluid Thioglycollate Medium 巯基醋酸盐液体培养基
重金属检查法 1.目的：建立重金属检查（药典中第一、二法）的标准规程。 2.范围：QC化验室。 3.责任：QC化验员。 4.内容： 4.1简述： pH值是3.0-3.5，选用醋酸盐缓冲液（pH3.5）2ml调节pH 较好，显色剂硫代乙酰胺试液用量经实验也以2ml为佳，显色时间一般为2分钟。以10-20μgPb与显色剂所产生的颜色为最佳目视比色范围。在规定实验条件下，与硫代乙酰胺试液在弱酸条件下产生的硫化氢呈色的金属离子有银、铅、汞、铜、镉、铋、锑、锡、砷、锌、钴与镍等。 4.2仪器与用具：纳氏比色管，应注意选择各管之间的平行 性，玻璃色泽一致，内径、刻度、标线高度一致。比色管洗涤时避免划伤内壁。 4.3试药和试液： ℃干燥至恒重的硝酸铅0.160g，置1000m1量瓶中，加硝酸5ml与水50ml溶解后，用水稀释至刻度，摇匀，作为贮备液。临用前，精密量取贮液10m1置100ml量瓶中，加
水稀释至刻度，摇匀，即得（每1ml相当于10μg的pb）。 4.4操作方法： pH3.5）2ml，加水或该药品项下规定的溶剂稀释成25ml。 μm）滤过，然后甲管中加入标准铅溶液一定量，水或该药品项下规定的溶剂使成25ml，再在乙管中加硫代乙酰胺试液2ml，甲管中加水2ml，照上述方法比较，即得。pH3.5）2ml与水15ml溶解以后，移置甲管中，加标准铅溶液一定量，再加水稀释成25ml。 ℃灼烧的炽灼残渣项下遗留的残渣，加硝酸0.5ml蒸干，至氧化氮蒸汽除尽后，放冷，加盐酸2ml，置水浴上蒸干后加水15m1，滴加氨试液至酚酞指示液显中性，再加醋酸盐缓冲液（pH3.5）2m1，微热溶解后，移至乙管中，加水稀释成25m1。 -1.0ml，使恰湿润，用低温加热至硫酸除尽后，加硝酸0.5ml，蒸干，至氧化氮蒸气除尽后，放冷，在500-600℃“…放冷，加盐酸2m1...”起，至加水稀释成25ml。 4.5注意事项： pH3.5）时，要用PH计调节，硫代乙酰胺试液加入量以2ml 时呈色最深；硫代乙酰胺试液显色剂的最佳显色时间为2分钟，第一、第二法均为放置2分钟。 μg的Pb）为宜。小于1ml或大于3m1，呈色太浅或太深均
《671》包装容器——性能检测 本章规定了用来包装的塑料容器及其组件功能性质上的标准(药品、生物制剂、营养补充剂和医疗器械)，定义了保存、包装、存储和标签方面的凡例与要求。本文提供的试验用于确定塑料容器的透湿性和透光率。盛装胶囊和片剂的多单元容器章节适用于多单元容器。盛装胶囊和片剂的单位剂量容器章节适用于单位剂量容器。盛装胶囊和片剂的多单元容器(没有密封) 的章节适用于没有密封的聚乙烯和聚丙烯容器。盛装液体的多元和单元容器的章节适用于多元的和单元的容器。 一个容器想要提供避光保护或作为一个符合耐光要求的容器，由具有耐光的特殊性质的材料组成，包括任何涂层应用。一个无色透明或半透明的容器通过一个不透明的外壳包装变成耐光的(见凡例和要求 )，可免于对光的透射要求。在多单元容器和封盖与水泡的单位剂量容器由衬垫密封情况下，此处使用的术语“容器”指的是整个系统的组成。 盛装胶囊和片剂的多元容器 干燥剂——放置一些颗粒4—8目的无水氯化钙在一个浅的容器里，仔细剔除细粉，然后置于110°干燥，并放在干燥器中冷却。 试验过程——挑选12个类型和尺寸一致的容器，用不起毛的毛巾清洁密闭表面，并打开和关闭每个容器30次。坚决每次应用容器密闭一致。通过扭矩关闭螺旋盖容器，使气密性在附表规定的范围内。10个指定的测试容器添加干燥剂，如果容器容积大于等于20mL，每个填充13mm以内封闭；如果容器的容积小于20毫升，每个填充容器容量的三分之二。如果容器内部的深度超过63mm，惰性填料或垫片可以放置在底部来最小化容器和干燥剂的总重量；干燥剂层在这样一个容器中深度不低于5cm。添加干燥剂之后，立即按附表中规定的扭矩封闭螺旋帽容器。剩余的2个指定为对照容器，每个添加足够数量的玻璃珠，重量约等于每个测试容器的重量，并用附表中规定的扭矩封闭螺旋帽容器。记录各个容器的重量，如果容器的容积小于20毫升，精确到0.1毫克；如果容器容积为20毫升或以上但小于200毫升，精确到毫克；如果容器容积为200毫升及以上，精确到厘克（10毫克）；在相对湿度75±3%和温度23±2°的环境下存储。[注意——浓度为35g/100mL的氯化钠溶液放在干燥器底部的渗透系统来维持指定湿度。其他的方法可以用来维护这些条件。] 336±1小时（14天）后，用同样的办法记录每个容器的重
USP35-NF-30结构整理 vivi2010-10-02 USP总目录： 1 New Official Text修订文件 加快修订过程包括勘误表，临时修订声明（IRAS），修订公告。勘误表，临时修订声明，修订公告在USP网站上New Official Text部分刊出，勘误表，临时修订公告也会在PF上刊出2front matter前言 药典与处方集增补删减情况，审核人员，辅料收录情况 3凡例
药典， 1标题和修订 2 药典地位和法律认可 3标准复合性 4专论和通则 5 专论组成 6 检验规范和检验方法 7 测试结果 8 术语和定义 9 处方和配药 10 包装存储与标签 4通则 4.1章节列表 4.2一般检查和含量测定（章节编号小于1000）
检查和含量分析的一般要求 检查和含量分析的仪器， 微生物检查，生物检查和含量测定， 化学检查和含量测定， 物理检查和测定 4.3一般信息（章节号大于1000） 5食物补充剂通则 6试剂（试剂，指示剂，溶液等） 7参考表 性状描述和溶解性查询表（按字母顺序） 8食品补充剂各论（字母顺序） 9NF各论（辅料标准） 10 USP各论 11术语 附件：通则的章节中文目录（使用起来比较方便，直接找对应章节号即可）一、通用试验和检定 （1）试验和检定的总要求 1 注射剂 11 参比标准物 （2）试验和检定的装置 16 自动分析方法 21 测温仪 31 容量装置，如容量瓶、移液管、滴定管，各种规格的误差限度
41 砝码和天平 （3）微生物学试验 51 抗菌效力试验 55 生物指示剂：耐受性能试验 61 微生物限度试验 61 非灭菌制品的微生物检查：计数试验 62 非灭菌制品的特定菌检查，如大肠杆菌、金葡菌、沙门氏菌等 71 无菌试验 （4）生物学试验和检定 81 抗生素微生物检定 85 细菌内毒素试验 87 体外生物反应性试验：检查合成橡胶、塑料、高聚物对哺乳类细胞培养的影响 88 体内生物反应性试验：检查上述物质对小鼠、兔iv、ip或肌内植入的影响 91 泛酸钙检定 111 生物检定法的设计和分析 115 右泛醇检定 121 胰岛素检定 141 蛋白质——生物适应试验，用缺蛋白饲料大鼠，观察水解蛋白注射液和氨基酸混合物的作用 151 热原检查法 161 输血、输液器及类似医疗装置的内毒素、热原、无菌检查 171 维生素B12 活性检定 （5）化学试验和检定 A 鉴别试验 181 有机含氮碱的鉴别 191 一般鉴别试验 193 四环素类鉴别 197 分光光度法鉴别试验 201 薄层色谱鉴别试验 B 限量试验
标准操作规程 Standard Operating Procedure 1.简述 1．1重金属是指在规定实验条件下能与硫代乙酰胺或硫化钠作用显色的金属盐类杂质。1．2硫化钠或硫代乙酰胺在弱酸性条件下水解产生硫化氢，与供试品中重金属在规定实验条件下所显颜色，与一定量的标准铅溶液在同样操作条件下所显的颜色比较。1．3由于实验条件不同。分为三种检查方法：第一法适用于在规定条件下能生成澄清无色溶液的供试品。第二法适用于在规定条件下不能生成澄清无色溶液的供试品；第三法适用于那些不能用一法和二法的样品。 2 仪器 2．1仪器设备 50ml纳氏比色管 2．2试剂和溶液 a)硫代乙酰胺试液: 称取硫代乙酰胺4g加水溶解，用水稀释至100ml，摇匀。 b)甘油基准试液: 称取200g甘油加水至总重量为235g,然后加142.5ml1N氢氧 化钠溶液和47.5ml的水。 临用前用0.2ml硫代乙酰胺试液和1ml甘油基准试液混合,在沸水浴中加热20秒钟,立即使用. c) 硝酸铅贮备液：称取硝酸铅0.1598g，置1000ml量瓶中，加硝酸1ml与水100ml
溶解后，用水稀释至1000ml 摇匀，作为贮备液。 标准铅溶液临用前，精密量取贮备液10ml，置100ml量瓶中，加水稀释至刻 SOP-QM 402-14 页号Page：2/4 度，摇匀，即得（每1ml相当于10μg的Pb）。 d）PH3.5醋酸盐缓冲液: 取醋酸铵25.0g，加水25ml溶解后，加38.0ml6N盐 酸,如有必要用6N氨水或6N盐酸调PH至3.5 ，用水稀释至 100ml，摇匀。 第一法： 标准溶液的制备取50ml比色管，用移液管移取2.0ml标准铅溶液(20 μgPb),用水稀释到25ml，用1N醋酸或6N氨水溶液调PH至3.0~4.0之间，用 窄范围的精密pH试纸作指示，然后加水稀释至40ml，摇匀。 供试品溶液的制备另取一支50ml比色管，加入按该品种项下规定的方法制成的供试液25ml；或加入供试品的量以g计, 按公式2.0/1000L计算,其中L 为重金属的限度(%)，用该品种项下规定的酸的体积溶解，加水溶解并稀释 至25ml，供试品溶液用1N醋酸或6N氨水溶液调PH至3.0～4.0之间，用窄 范围的精密pH试纸作指示,然后加水稀释至40ml，摇匀。 对照溶液的制备取第三只比色管，加入与上述供试品溶液方法同样制备的任意25ml，加入2.0ml标准铅溶液，用1N醋酸或6N氨水溶液调PH至3.0~4.0 之间，用窄范围的精密pH试纸作指示,然后加水稀释至40ml，摇匀。 操作方法在上述三只比色管中分别加入2ml醋酸缓冲溶液（PH3.5）, 然后 加1.2ml硫代乙酰胺_甘油基准试剂，加水稀释到50ml，摇匀，放置2分钟，同置白色 背景下，至上向下透视.供试品溶液的颜色不得深于标准溶液的颜色；对照溶液的颜色 相同于或较深于标准溶液的颜色( 注:如对照液的颜色浅于标准品溶液,则以第二法代 替第一法)。 第二法： pH3.5醋酸盐缓冲液照第一法配制 标准溶液的制备用移液管取4ml标准铅溶液到适宜的试管中，并加入10ml 6N盐酸。 照方法一配制 供试品溶液的制备称取一定量的样品(以g计算) ,按公式4.0/1000L 2.0/1000L计
á233? ELEMENTAL IMPURITIES—PROCEDURES INTRODUCTION This chapter describes two analytical procedures (Procedures 1 and 2) for the evaluation of the levels of the elemental impuri-ties. The chapter also describes criteria for acceptable alternative procedures. By means of validation studies, analysts will confirm that the analytical procedures described herein are suitable for use on specified material. Use of Alternative Procedures The chapter also describes criteria for acceptable alternative procedures. Alternative procedures that meet the validation re-quirements herein may be used in accordance with General Notices and Requirements 6.30, Alternative and Harmonized Meth-ods and Procedures . Information on the Requirements for Alternate Procedure Validation is provided later in this chapter.Speciation The determination of the oxidation state, organic complex, or combination is termed speciation . Analytical procedures for spe-ciation are not included in this chapter, but examples may be found elsewhere in USP–NF and in the literature. PROCEDURES ? C OMPENDIAL P ROCEDURES 1 AND 2 System standardization and suitability evaluation using applicable reference materials should be performed on the day of analysis. Procedure and detection technique:Procedure 1 can be used for elemental impurities generally amenable to detection by inductively coupled plasma–atomic (optical) emission spectroscopy (ICP–AES or ICP–OES). Procedure 2 can be used for ele-mental impurities generally amenable to detection by ICP–MS. Before initial use, the analyst should verify that the proce- dure is appropriate for the instrument and sample used (procedural verification) by meeting the alternative procedure vali-dation requirements below. Sample preparation:Forms of sample preparation include Neat , Direct aqueous solution , Direct organic solution , and Indi- rect solution . The selection of the appropriate sample preparation depends on the material under test and is the responsibil-ity of the analyst. When a sample preparation is not indicated in the monograph, an analyst may use any of the following appropriately validated preparation procedures. In cases where spiking of a material under test is necessary to provide an acceptable signal intensity, the blank should be spiked with the same Target elements , and where possible, using the same spiking solution. Standard solutions may contain multiple Target elements . [N OTE —All liquid samples should be weighed.]Neat:Used for liquids or alternative procedures that allow the examination of unsolvated samples. Direct aqueous solution:Used when the sample is soluble in an aqueous solvent. Direct organic solution:Used where the sample is soluble in an organic solvent. Indirect solution:Used when a material is not directly soluble in aqueous or organic solvents. Total metal extraction is the preferred sample preparation approach to obtain an Indirect solution . Digest the sample using the Closed vessel diges-tion procedure provided below or one similar to it. The sample preparation scheme should yield sufficient sample to allow quantification of each element at the limit specified in the corresponding monograph or chapter. Closed vessel digestion:This sample preparation procedure is designed for samples that must be digested in a Concen-trated acid using a closed vessel digestion apparatus. Closed vessel digestion minimizes the loss of volatile impurities. The choice of a Concentrated acid depends on the sample matrix. The use of any of the Concentrated acids may be appropri-ate, but each introduces inherent safety risks. Therefore, appropriate safety precautions should be used at all times. [N OTE —Weights and volumes provided may be adjusted to meet the requirements of the digestion apparatus used.] An example procedure that has been shown to have broad applicability is the following. Dehydrate and predigest 0.5 g of primary sample in 5 mL of freshly prepared Concentrated acid . Allow to sit loosely covered for 30 min in a fume hood.Add an additional 10 mL of Concentrated acid , and digest, using a closed vessel technique, until digestion or extraction is complete. Repeat, if necessary, by adding an additional 5 mL of Concentrated acid . [N OTE —Where closed vessel digestion is necessary, follow the manufacturer’s recommended procedures to ensure safe use.] Alternatively, leachate extraction may be appropriate with justification following scientifically validated metal disposition studies, which may include animal studies, speciation, or other means of studying disposition of the specific metal in the drug product. Reagents:All reagents used for the preparation of sample and standard solutions should be free of elemental impurities,in accordance with Plasma Spectrochemistry á730?. ? P ROCEDURE 1: ICP–OES Standardization solution 1: 1.5J of the Target element(s) in a Matched matrix Standardization solution 2:0.5J of the Target element(s) in a Matched matrix Sample stock solution:Proceed as directed in Sample preparation above. Allow the sample to cool, if necessary. For mer-cury determination, add an appropriate stabilizer. Sample solution:Dilute the Sample stock solution with an appropriate solvent to obtain a final concentration of the Target elements at NMT 1.5J . Blank: Matched matrix 298 á233? Elemental Impurities—Procedures / Chemical Tests USP 40
921WATER DETERMINATION水分测定 Many Pharmacopeial articles either are hydrates or contain water in adsorbed form. As a result, the determination of the water content is important in demonstrating compliance with the Pharmacopeial standards. Generally one of the methods given below is called for in the individual monograph, depending upon the nature of the article. In rare cases, a choice is allowed between two methods. When the article contains water of hydration, the Method I (Titrimetric), the Method II (Azeotropic), or the Method III (Gravimetric) is employed, as directed in the individual monograph, and the requirement is given under the heading Water. 很多药典物品要么是水合物，要么含有处于吸附状态的水。因此，测定水分含量对于证实与药典标准的符合性是很重要的。通常，在具体的各论中会根据该物品的性质，要求使用下面若干方法中的一个。偶尔，会允许在2个方法中任选一个。当该物品含有水合状态的水，按照具体各论中的规定，使用方法I（滴定测量法）、方法II（恒沸测量法）、或方法III（重量分析法），这个要求在标题水分项下给出。 The heading Loss on drying (see Loss on Drying 731) is used in those cases where the loss sustained on heating may be not entirely water. 在加热时的持续失重可能不全是水分的情况下，使用标题干燥失重（见干燥失重<731>）。 METHOD I (TITRIMETRIC) 方法I（滴定测量法） Determine the water by Method Ia, unless otherwise specified in the individual monograph. 除非具体各论中另有规定，使用方法Ia来测定水分。 Method Ia (Direct Titration) 方法Ia（直接滴定） Principle— The titrimetric determination of water is based upon the quantitative reaction of water with an anhydrous solution of sulfur dioxide and iodine in the presence of a buffer that reacts with hydrogen ions. 原理：水分的滴定法检测是基于水与二氧化硫的无水溶液以及存在于缓冲液中与氢离子反应的碘之间的定量反应。 In the original titrimetric solution, known as Karl Fischer Reagent, the sulfur dioxide and iodine are dissolved in pyridine and methanol. The test specimen may be titrated with the Reagent directly, or the analysis may be carried out by a residual titration procedure. The stoichiometry of the reaction
L1和L8是美国药典(USP)规定的色谱柱编号，其实就是C18柱和NH2柱。下面是对应的色谱柱类型。 L1：十八烷基键合多孔硅胶或无机氧化物微粒固定相，简称C18或ODS L2：30～50um表面多孔薄壳型键合C18(ODS)固定相 L3：多孔硅胶微粒即一般的硅胶柱 L4：30～50um表面多孔薄壳型硅胶 L5：30～50um表面多孔薄壳型氧化铝 L6：30～50um实心微球表面包覆磺化碳氟聚合物－强阳离子交换固定相 L7：全多孔硅胶微粒键合C8官能团固定相简称C8柱 L8：全多孔硅胶微粒键合非交联NH2固定相简称NH2柱 L9：强酸性阳离子交换基团键合全多孔不规则形硅胶固定相 L10：多孔硅胶微球键合氰基固定相（CN）简称CN柱 L11：键合苯基多孔硅胶微球固定相简称苯基柱 L12：无孔微球键合季胺功能团的强阴离子填料 L13：三乙基硅烷化学键合全多孔硅胶微球固定相（C1）简称C1柱 L14:10um硅胶化学键合强碱性季铵盐阴离子交换固定相简称SAX柱 L15：已基硅烷化学键合全多孔硅胶微球固定相简称C6柱 L16：二甲基硅烷化学键合全多孔硅胶微粒固定相 L17：氢型磺化交联苯乙烯-二乙烯基苯共聚物,强阳离子交换树脂 L18: 3～10um全多孔硅胶化学键合胺基（NH2）和氰基（CN） L19：钙型磺化交联苯乙烯－二乙烯基苯共聚物，强阳离子交换树脂 L20：二羟基丙烷基化学键合多孔硅胶微球固定相（Diol）简称二醇基柱 L21：刚性苯乙烯－二乙烯基苯共聚物微球 L22：带有磺酸基团的多孔苯乙烯阳离子交换树脂 L23：带有季胺基团的聚甲基丙烯酸甲酯或聚丙烯酸酯多孔离子交换树脂 L24：表面含有大量羟基的半刚性聚乙烯醇亲水凝胶 L25：聚甲基丙烯酸酯树脂交联羟基醚（表面含有残余羧基功能团）树脂。能分离分子量100～5000MW范围的水溶性中性、阳离子型及阴离子型聚合物（用聚氧乙烯测定）的固定相 L26：丁基硅烷化学键合全多孔硅胶微球固定相 L27：30～50um的全多孔硅胶微粒