【正文】 ing the preparation being examined to which the standard endotoxin has been added and which has then been submitted to the chosen treatment.可通過其他適宜的方法（如過濾、中和、透析或加熱處理等）排除干擾。Table Solution Endotoxin concentration/Solution to which endotoxin is addedNumber of replicatesANone/Diluted test solution2B2λ/Diluted test solution2C2λ/Water for BET2DNone/Water for BET2溶液內(nèi)毒素濃度/配制內(nèi)毒素的溶液平行管數(shù)A0/供試品溶液2B2λ/供試品溶液2C2λ/BET用水2D0/BET用水2Prepare solution A and solution B (positive product control) using a dilution not greater than the MVD and treatments as described in 1. Preparatory testing, (ii) Test for interfering factors. Solutions B and C (positive controls) contain the standard endotoxin at a concentration corresponding to twice the labeled lysate sensitivity. Solution D (negative control) consists of water for BET.制備溶液A、B（供試品陽性對(duì)照），稀釋倍數(shù)不得超過MVD。(ii) Interpretation結(jié)果判斷The test is considered valid when both replicates of solution B and C are positive and those of solution D are negative.只有溶液B和C的平行管的試驗(yàn)結(jié)果均為陽性、溶液D的平行管的試驗(yàn)結(jié)果均為陰性時(shí)，試驗(yàn)方有效。In the repeat test, the preparation being examined plies with the test if a negative result is found for both replicates of solution A. The preparation does not ply with the test if a positive result is found for one or both replicates of solution A.在重復(fù)試驗(yàn)中，溶液A的平行管的試驗(yàn)結(jié)果均為陰性時(shí)，可判定供試品符合試驗(yàn)的內(nèi)毒素限度要求。、B、C、D。也可以準(zhǔn)備其它合適倍數(shù)的稀釋液。供試品的內(nèi)毒素濃度指這些平行管的反應(yīng)終點(diǎn)濃度的幾何平均值。如供試品內(nèi)毒素的濃度小于其專論中的規(guī)定濃度，可判定供試品符合檢查要求。The kineticturbidimetric test (Method C) is a method to measure either the time (onset time) needed for the reaction mixture to reach a predetermined absorbance or transmission, or the rate of turbidity development.動(dòng)態(tài)濁度法（方法C）是檢測(cè)反應(yīng)混合物的濁度達(dá)到某一預(yù)先設(shè)定的吸光度或透光率需要的反應(yīng)時(shí)間（起效時(shí)間），或是檢測(cè)濁度變化率。1℃）。The kineticchromogenic test (Method D) measures either the time (onset time) needed for the reaction mixture to reach a predetermined absorbance, or the rate of colour development.The test is carried out at the incubation temperature remended by the lysate manufacturer (usually 37 177。1℃）。用內(nèi)毒素標(biāo)準(zhǔn)溶液制成至少三個(gè)濃度的稀釋液，以獲取標(biāo)準(zhǔn)曲線。 (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(yàn)選擇標(biāo)準(zhǔn)曲線中點(diǎn)或一個(gè)靠近中點(diǎn)的內(nèi)毒素濃度。 溶液B=加入了標(biāo)準(zhǔn)曲線終點(diǎn)或靠近中點(diǎn)的一個(gè)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。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(biāo)準(zhǔn)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(jì)算內(nèi)毒素的平均回收率。重復(fù)干擾因素實(shí)驗(yàn)以驗(yàn)證處理的有效性。(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(jì)算用系列溶液C生成的標(biāo)準(zhǔn)曲線計(jì)算溶液A的每一個(gè)平行管的內(nèi)毒素濃度。(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(jì)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(xì)菌內(nèi)毒素檢查的要求。(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(jì)算出的回收率在50％－200％的范圍內(nèi)。為確保所選擇的處理方法能有效地排除干擾且不會(huì)使內(nèi)毒素失去活性，要使用預(yù)先添加了標(biāo)準(zhǔn)內(nèi)毒素再經(jīng)過處理的供試品溶液進(jìn)行干擾實(shí)驗(yàn)。在溶液中不添加任何內(nèi)毒素的條件下，計(jì)算任何所測(cè)內(nèi)毒素的減少。 溶液D=BET用水（陰性對(duì)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 。在鱟試劑生產(chǎn)商的建議測(cè)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(shí)間等）下，對(duì)這些溶液開展試驗(yàn)，每種溶液至少做兩組平行。使用動(dòng)態(tài)法檢測(cè)時(shí)，如預(yù)期范圍超過2 log，為使標(biāo)準(zhǔn)曲線反映出每個(gè)對(duì)數(shù)增長(zhǎng)，應(yīng)相應(yīng)增加標(biāo)準(zhǔn)品溶液。當(dāng)實(shí)驗(yàn)條件發(fā)生了任何可能會(huì)影響檢驗(yàn)結(jié)果的改變時(shí)，需要驗(yàn)證試驗(yàn)方法。C).動(dòng)態(tài)顯色法（方法D）是檢測(cè)反應(yīng)混合物的色度達(dá)到某一預(yù)定吸光度（或透光率）所需要的反應(yīng)時(shí)間（起效時(shí)間），或檢測(cè)色度增長(zhǎng)速度的方法。根據(jù)檢測(cè)量之間存在的量化關(guān)系來測(cè)定內(nèi)毒素含量的方法。 1 176。根據(jù)檢測(cè)原理，這種方法又可分為終點(diǎn)濁度法和動(dòng)態(tài)濁度法。如在有效的試驗(yàn)中，供試品的檢查結(jié)果均顯陰性，應(yīng)記為內(nèi)毒素濃度小于λ（如實(shí)驗(yàn)使用的是供試品的稀釋液，檢查結(jié)果應(yīng)記為內(nèi)毒素濃度小于λ乘以供試品的最小稀釋倍數(shù)）。溶液D=BET用水（陰性對(duì)照）(ii) Calculation and interpretationThe test is considered valid when the following 3 conditionsare met:(a) both replicates of solution D (negative control) are negative,(b) both replicates of solution B (positive product control)are positive,(c) the geometric mean endpoint concentration of solution Cis in the range of to 2λ.(ii)計(jì)算和結(jié)果判斷符合下列三個(gè)條件時(shí)，可將試驗(yàn)判為有效：（a） 溶液D（陰性對(duì)照）的兩組平行管均顯陰性，（b）（b）溶液B（供試品陽性對(duì)照）的兩個(gè)平行管均顯陽性，（c）—2λ之間。SolutionEndotoxin Concentration/Solution to which Endotoxin is AddedDiluentDilution FactorInitial Endotoxin ConcentrationNumber of ReplicatesAanone/sample solutionWater for BET 1248— ———2222Bb2/sample solution—122Cc2/ Water for BETWater for BET 12482 12222Ddnone/ Water for BET———2Solution A = test solution at the dilution, not exceeding the MVD, with which the test for interfering factors was carried out.