【正文】 of the correlation coefficient, | r |, must be greater than or equal to , for the range of endotoxin concentrations set up.（i） 標準曲線的可靠性試驗每批鱟試劑都必須進行此項試驗。用內(nèi)毒素標準溶液制成至少三個濃度的稀釋液，以獲取標準曲線。按照鱟試劑生產(chǎn)商的建議（體積比、孵育時間、溫度、pH等），每一內(nèi)毒素標準濃度的溶液至少做三支平行管。使用動態(tài)法檢測時，如預期范圍超過2 log，為使標準曲線反映出每個對數(shù)增長，應相應增加標準品溶液。在鱟試劑生產(chǎn)商規(guī)定的內(nèi)毒素濃度范圍內(nèi)，相關(guān)系數(shù)的絕對值︱r︱。 (ii)Test for interfering factors Select an endotoxin concentration at or near the middle of the endotoxin standard curve.Prepare solutions A, B, C and D as shown in Table . Perform the test on at least 2 replicates of these solutions as remended by the lysate manufacturer (volume of test solution and lysate solution, volume ratio oftest solution to lysate solution, incubation time, etc.)(ii)干擾因素試驗選擇標準曲線中點或一個靠近中點的內(nèi)毒素濃度。、B、C、D。在鱟試劑生產(chǎn)商的建議測試條件（供試品和鱟試液體積、供試品于鱟試液的體積比、孵育時間等）下，對這些溶液開展試驗，每種溶液至少做兩組平行。SolutionEndotoxin ConcentrationSolution to which Endotoxin is AddedNumber of ReplicatesAanonesample solutionnot less than 2Bbmiddle concentration of the standard curvesample solutionnot less than 2Ccat least 3 concentrations (lowest concentration is designated )Water for BET each concentration not less than 2DdnoneWater for BET not less than 2Solution A = test solution, that may be diluted not to exceed the MVD.Solution B = preparation to be examined at the same dilution assolution A, containing added endotoxin at a concentration equal to ornear the middle of the standard curve.Solution C = standard endotoxin solution at the concentrations used inthe validation of the method as described under 3. Preparatory testing,(i) Assurance of criteria for the standard curve (positive controls).Solution D = water for BET (negative control). 溶液內(nèi)毒素濃度配制內(nèi)毒素的溶液平行管或微孔數(shù)A無供試品溶液不少于2個B標準曲線的中點供試品溶液不少于2個C至少3個濃度（最低濃度規(guī)定為λ）BET檢查每一濃度不少于2個D無BET檢查用水不少于2個 溶液A=稀釋倍數(shù)不超過MVD的供試品溶液。 溶液B=加入了標準曲線終點或靠近中點的一個已知濃度內(nèi)毒素的、且與溶液A有相同稀釋倍數(shù)的供試品溶液。 溶液C=如“（i）標準曲線的可靠性試驗”項下描述的、用于制備標準曲線的標準內(nèi)毒素溶液。 溶液D=BET用水（陰性對照）The test is considered valid when the following conditionsare met:– the absolute value of the correlation coefficient of the standard curve generated using solution C is greater than or equal to 。– the result with solution D does not exceed the limit of the blank value required in the description of the lysate reagent employed, or it is less than the endotoxin detection limit of the lysate reagent employed.試驗必須符合以下條件方有效： ； 溶液D（陰性對照）的試驗結(jié)果不得大于所用鱟試劑的說明書中規(guī)定的空白值的限度，或小于鱟試劑的內(nèi)毒素檢測限。Calculate the mean recovery of the added endotoxin by subtracting the mean endotoxin concentration in the solution (if any) (solution A, Table ) from that in the solution containing the added endotoxin (solution B, Table ).用含標準內(nèi)毒素的供試品溶液（溶液B，）的平均內(nèi)毒素含量減去供試品溶液（溶液A，）的平均內(nèi)毒素含量，計算內(nèi)毒素的平均回收率。The test solution is considered free of interfering factors if under the conditions of the test, the measured concentration of the endotoxin added to the test solution is within 50200 per cent of the known added endotoxin concentration, after subtraction of any endotoxin detected in the solution without added endotoxin. 當內(nèi)毒素的回收率在50％－200％之間時，可認為在此試驗條件下供試品溶液中不存在干擾因素。在溶液中不添加任何內(nèi)毒素的條件下，計算任何所測內(nèi)毒素的減少。When the endotoxin recovery is out of the specified range,the test solution is considered to contain interfering factors. Repeat the test using a greater dilution, not exceeding the MVD. Furthermore, interference of the test solution or diluted test solution not to exceed the MVD may be eliminated by suitable validated treatment, such as filtration, neutralisation, dialysis or heat treatment. To establish that the treatment chosen effectively eliminates interference without loss of endotoxins, repeat the test for interfering factors using the preparation being examined to which the standard endotoxin has been added and which has then been submitted to the chosen treatment.當內(nèi)毒素的回收率不在指定范圍內(nèi)，應根據(jù)凝膠法章節(jié)的“（ii）干擾因素實驗”項下的說明排除干擾因素。重復干擾因素實驗以驗證處理的有效性。此外，可通過其他適宜的方法（如過濾、中和、透析或加熱處理等）排除供試液或未超過MVD的稀釋供試液的干擾。為確保所選擇的處理方法能有效地排除干擾且不會使內(nèi)毒素失去活性，要使用預先添加了標準內(nèi)毒素再經(jīng)過處理的供試品溶液進行干擾實驗。4. TEST檢查法(i) ProcedureFollow the procedure described in 3. Preparatory testing,(ii) Test for interfering factors.按 “（3）預備試驗下的（ii）干擾因素試驗”項下操作步驟。(ii) CalculationCalculate the endotoxin concentration of each replicate of solution A using the standard curve generated by the positive control solution test is considered valid when the following 3 requirements are met:(1) the results obtained with solution C ply with the requirements for validation defined under 3. Preparatory testing, (i) Assurance of criteria for the standard curve,(2) the endotoxin recovery, calculated from the endotoxin concentration found in solution B after subtracting the endotoxin concentration found in solution A, is within the range of 50200 per cent,(3) the result obtained with solution D (negative control) does not exceed the limit of the blank value required in the description of the lysate employed, or it is less than the endotoxin detection limit of the lysate reagent employed.(ii)計算用系列溶液C生成的標準曲線計算溶液A的每一個平行管的內(nèi)毒素濃度。試驗必須符合以下三個條件方有效：陽性對照系列溶液C的檢查結(jié)果要符合“(1)實驗（i）標準曲線的可靠性試驗”中的要求。(2)用溶液B中的內(nèi)毒素濃度減去溶液A中的內(nèi)毒素濃度后，計算出的回收率在50％－200％的范圍內(nèi)。(3)溶液D（陰性對照）的試驗結(jié)果不得大于所用鱟試劑的說明書中規(guī)定的空白值的限度或小于鱟試劑的內(nèi)毒素檢測限。(iii) InterpretationThe preparation being examined plies with the test if the mean endotoxin concentration of the replicates of solution A,after correction for dilution and concentration, is less than the endotoxin limit for the on the test for bacterial endotoxins are given in general chapter .(iii)結(jié)果判斷若校正了稀釋倍數(shù)和濃度后，由溶液A的內(nèi)毒素濃度計算得到的供試品內(nèi)毒素平均濃度小于供試品的內(nèi)毒素限度，可判斷供試品符合細菌內(nèi)毒素檢查的要求。.內(nèi)容總結(jié)
（1）NOTE: in this chapter, the term ‘tube’ includes all types of receptacles, for example microtitre plate wells.
注：這一章中，“管”的意思包括其他任何反應容器，如微孔板中的孔