2.9.19. PARTICULATE CONTAMINATION: SUB-VISIBLE PARTICLES

2.9.19. 顆粒物污染：亞可見顆粒物檢(jiǎn)查規(guī)定

Particulate contamination of injections and infusions consists of extraneous, m o bile undissolved particles, other than gas bubbles, unintentionally present in the solutions.

注射劑(jì)和輸液的微粒污染包括在溶液中無意中存在的外來的、可移動(dòng)的未溶解顆粒，而不是氣泡。

For the determination of particulate contamination 2 procedures, Method 1 (Light Obscuration Particle Count Test) and Method 2 (Microscopic Particle Count Test), are specified hereinafter. When examining injections and infusions for sub-visible particles, Method 1 is preferably applied. However, it may be necessary to test some preparations by the light obscuration particle count test followed by the microscopic particle count test to reach a conclusion on conformance to the requirements.

對於(yú)顆粒物污染的測定包含兩種方法，下文進行瞭(le)詳細介紹：方法1（光阻法顆(kē)粒計(jì)數試驗）和方法2（顯微顆粒物計數試驗）。當(dāng)檢查注射劑(jì)和輸液中是否存在亞可見顆粒時，最好使用方法1。然而，可能有必要通過(guò)光阻法顆粒計數試驗和顯微顆粒計數試驗來測(cè)試某些制劑，以得出符合要求的結論。

Not all parenteral preparations can be examined for sub-visible particles by one or both of these methods. When Method 1 is not applicable, e.g. in case of preparations having reduced clarity or increased viscosity, the test is carried out according to Method 2. Emulsions, colloids, and liposomal preparations are examples. Similarly, products that produce air or gas bubbles when drawn into the sensor may also require microscopic particle count testing. If the viscosity of the preparation to be tested is sufficiently high so as to preclude its examination by either test method, a quantitative dilution with an appropriate diluent may be made to decrease viscosity, as necessary, to allow the analysis to be performed.

並(bìng)非所有非腸(cháng)道制劑都可以通過這些方法中的一種或兩種方法檢查亞可見顆粒。例如在制劑的透明度降低或粘度增加的情況下，方法 1 不适用時，應根據方法2進行測(cè)試，如乳液、膠體和脂質體制劑(jì)等。 同樣
，在吸入傳感器時會産(chǎn)生空氣或氣泡的産(chǎn)品也可能需要進行顯微顆粒物計數測(cè)試。 如果待測(cè)制劑的粘度足夠高以至於(yú)無法通過任一測(cè)試方法對其進行檢查，則可以根據需要，用适當的稀釋劑進行定量稀釋以降低粘度，以便進行分析。

The results obtained in examining a discrete unit or group of units for particulate contamination cannot be extrapolated with certainty to other units that remain untested. Thus, statistically sound sampling plans must be developed if valid inferences are to be drawn from observed data to characterize the level of particulate contamination in a large group of units.

在檢查一個離散單(dān)元或一組單(dān)元的微粒污染時獲得的結果不能肯定地外推到其他未測(cè)試的單(dān)元。 因此，如果要從(cóng)觀察到的數據中得出有效的推論，以表征一大組單(dān)位中的顆粒污染水平，就必須制定統計上合理的抽樣計劃。

METHOD 1. LIGHT OBSCURATION PARTICLE COUNTTEST

光阻法顆粒計數實驗

Use a suitable apparatus based on the principle of light blockage which allows an automatic determination of the size of particles and the number of particles according to size. The apparatus is calibrated using suitable certified reference materials consisting of dispersions of spherical particles of known sizes between 10 µm and 25 µm. These standard particles are dispersed in particle-free water R. Care must be taken to avoid aggregation of particles during dispersion.

根據光阻原理，使用合适的儀器，可以根據大小自動(dòng)確(què)定顆粒大小和顆粒數量。使用合适的認證标準物質對儀器進行校準，該标準物質由已知尺寸在10 µm到25 µm之間(jiān)的球形顆粒的分散體組成。這些标準顆粒分散在無顆粒水中。分散過(guò)程中必須注意避免顆粒聚集。

General precautions 一般注意事項

The test is carried out under conditions limiting particulate contamination, preferably in a laminar-flow cabinet. Very carefully wash the glassware and filtration equipment used, except for the membrane filters, with a warm detergent solution and rinse with abundant amounts of water to remove all traces of detergent. Immediately before use, rinse the equipment from top to bottom, outside and then inside, with particle-free water R.

實驗在限制顆粒污染的條件下進行，最好在層流櫃中進行。使用溫熱的清潔劑溶液仔細清洗所用的玻璃器皿和過濾設備(bèi)（膜式過濾器除外），並(bìng)用大量水沖洗，以去除所有清潔劑痕迹。使用前，用無顆粒水從上到下、從外到内沖洗設備(bèi)。

Take care not to introduce air bubbles into the preparation to be examined, especially when fractions of the preparation are being transferred to the container in which the determination is to be carried out.

注意不要将氣泡引入待檢查的制劑(jì)中，尤其是将制劑(jì)的部分轉移到要進行測(cè)定的容器中時。

In order to check that the environment is suitable for the test, that the glassware is properly cleaned and that the water to be used is particle-free, the following test is carried out: determine the particulate contamination of 5 samples of particle-free water R, each of 5 mL, according to the method described below. If the number of particles of 10 µm or greater size exceeds 25 for the combined 25 mL, the precautions taken for the test are not sufficient. The preparatory steps must be repeated until the environment, glassware and water are suitable for the test。

爲瞭(le)檢查環境是否适合試驗，玻璃器皿是否正確(què)清潔，所使用的水是否無顆粒，應進行以下試驗：根據下述方法，測定5個(gè)無顆(kē)粒水樣品的顆(kē)粒污染情況，每個(gè)樣品5 mL。如果10 µm或更大粒徑的顆粒數量超過25個，則爲試驗採(cǎi)取的預防措施是不夠的。必須重複準備(bèi)步驟，直到環境、玻璃器皿和水适合試驗

Method方法

Mix the contents of the sample by slowly inverting the container 20 times successively. If necessary, cautiously remove the sealing closure. Clean the outer surfaces of the container opening using a jet of particle-free water R and remove the closure, avoiding any contamination of the contents. Eliminate gas bubbles by appropriate measures such as allowing to stand for 2 min or sonicating.

通過連續20次緩慢翻轉容器，混合樣品裏的内容物。如有必要，小心地拆下密封蓋。使用無顆粒水水流清潔容器開口的外表面，並(bìng)拆下封蓋，避免内容物受到任何污染。通過适當(dāng)的措施消除氣泡，例如靜置2分鍾或超聲波處理。

For large-volume parenterals, single units are tested. For small-volume parenterals less than 25 mL in volume, the contents of 10 or more units are combined in a cleaned container to obtain a volume of not less than 25 mL; where justified and authorised, the test solution may be prepared by mixing the contents of a suitable number of vials and diluting to 25 mL with particle-free water R or with an appropriate solvent without contamination of particles when particle-free water R is not suitable. Small-volume parenterals having a volume of 25 mL or more may be tested individually.

對於(yú)大容量腸外注射劑，需要測(cè)試單個單元。對於(yú)體積小於(yú)25 mL的小容量腸外注射劑，将10個(gè)或更多個(gè)的内容物組合在一個(gè)清潔的容器中，以獲得不小於(yú)25 mL的體積的溶液；在合理和授權的情況下，可通過混合适當(dāng)數量的小瓶中的内容物，並(bìng)用無顆粒水稀釋至25 mL，或在無顆粒水不适用時，用無顆粒污染的适當(dāng)溶劑(jì)稀釋至25 mL來制備試驗溶液。體積爲25 mL或以上的小容量非腸(cháng)道藥物試劑可單(dān)獨進行試驗。

Powders for parenteral administration are reconstituted with particle-free water R or with an appropriate solvent without contamination of particles when particle-free water R is not suitable.

用於(yú)腸(cháng)外給藥的粉末用無顆粒水重新配制，或當無顆粒水不适用時，使用适當的無顆粒污染的溶劑重新進行配制。

The number of test specimens must be adequate to provide a statistically sound assessment. For large-volume parenterals or for small-volume parenterals having a volume of 25 mL or more, fewer than 10 units may be tested, based on an appropriate sampling plan。

測(cè)試樣本的數量必須足夠，以提供統計上合理的評估。根據适當的抽樣計劃，對於(yú)大容量或25 mL以上的小容量腸外靜脈，可檢測(cè)小於(yú)10個樣品

Remove 4 portions, each of not less than 5 mL, and count the number of particles equal to or greater than 10 µm and 25 µm. Disregard the result obtained for the first portion, and calculate the mean number of particles for the preparation to be examined.

取下4份樣品，每份不少於5 mL，並計算等於或大於10 µm和25 µm的顆粒數。忽略第一份的結果，並(bìng)計算待檢查制劑(jì)的平均顆粒數。

Evaluation評估

For preparations supplied in containers with a nominal volume of more than 100 mL, apply the criteria of test 1.A.

For preparations supplied in containers with a nominal volume of less than 100 mL, apply the criteria of test 1.B.

♦For preparations supplied in containers with a nominal volume of 100 mL, apply the criteria of test 1.B. ♦

If the average number of particles exceeds the limits, test the preparation by the microscopic particle count test.

對於在标稱體積超過100 mL的容器中提供的制劑(jì)
，應採(cǎi)用試驗1A的标準。

對於在标稱體積小於100 mL的容器中提供的制劑(jì)，採(cǎi)用試驗1B的标準。

對於在标稱體積爲100 mL的容器中提供的制劑(jì)，採(cǎi)用試驗1B的标準。

如果平均顆粒數超過限值，則通過顯微顆粒計數測(cè)試對(duì)制劑進行測(cè)試。

Test 1.A – Solutions for infusion or solutions for injection supplied in containers with a nominal content of more than 100 mL

The preparation complies with the test if the average number of particles present in the units tested does not exceed 25 per millilitre equal to or greater than 10 µm and does not exceed 3 per millilitre equal to or greater than 25 µm.

試驗1A–輸液溶液或注射溶液，裝在标稱(chēng)含量超過(guò)100 mL的容器中。

如果被測(cè)單位樣品中粒徑等於(yú)或大於(yú)10 µm的顆(kē)粒的平均數量不超過(guò)每毫升25個，且粒徑等於或大於25 µm的顆(kē)粒平均數量不超過(guò)每毫升3個(gè)，則制劑(jì)符合試驗要求。

Test 1.B – Solutions for infusion or solutions for injection supplied in containers with a nominal content of less than 100 mL

The preparation complies with the test if the average number of particles present in the units tested does not exceed 6000 per container equal to or greater than 10 µm and does not exceed 600 per container equal to or greater than 25 µm.

試驗1B–輸液溶液或注射溶液，裝在标稱(chēng)含量小於(yú)100 mL的容器中。

如果被測(cè)單元中粒徑等於(yú)或大於(yú)10 µm的顆粒的平均數量不超過6000個/容器，且粒徑等於或大於25 µm的顆粒平均數量不超過600個/容器，則制劑(jì)符合試(shì)驗要求。

METHOD 2. MICROSCOPIC PARTICLE COUNT TEST

顯微顆粒計數

Use a suitable binocular microscope, filter assembly for retaining particulate contamination and membrane filter for examination.

使用合适的雙目顯微鏡、過(guò)濾器組件（用於(yú)保留顆粒污染物）和薄膜過(guò)濾器（用於(yú)檢查）。

The microscope is equipped with an ocular micrometer calibrated with an objective micrometer, a mechanical stage capable of holding and traversing the entire filtration area of the membrane filter, 2 suitable illuminators to provide episcopic illumination in addition to oblique illumination, and is adjusted to 100 ± 10 magnifications.

該顯微鏡配有一個目鏡測(cè)微計（使用物鏡測(cè)微計校準）、一個機械工作台（能夠容納並(bìng)穿過膜過濾器的整個過濾區域）、2個合适的照明器（除瞭(le)傾斜照明外，還提供反射照明），並(bìng)将其調整爲100±10倍。

The ocular micrometer is a circular diameter graticule (see Figure 2.9.19.-1.) and consists of a large circle divided by crosshairs into quadrants, transparent and black reference circles 10 µm and 25 µm in diameter at 100 magnifications, and a linear scale graduated in 10 µm increments. It is calibrated using a stage micrometer that is certified by either a domestic or international standard institution. A relative error of the linear scale of the graticule within ± 2 per cent is acceptable. The large circle is designated the graticule field of view (GFOV).

目鏡測(cè)微計是一個(gè)圓形直徑分劃器（見圖2.9.19.-1）由一個(gè)被十字準線(xiàn)分成四個(gè)象限的大圓、100倍放大時直徑爲10 µm和25 µm的透明和黑色參(cān)考圓(yuán)
、以及以10 µm增量刻度的線性刻度組成。使用經過國内或國際标準機構認證的測(cè)微計對(duì)其進行校準。分劃線性刻度的相對(duì)誤差在±2%以内是可以接受的。大圓被designated爲網格視場（GFOV）。

2 illuminators are required. One is an episcopic brightfield illuminator internal to the microscope, the other is an external, focusable auxiliary illuminator adjustable to give reflected oblique illumination at an angle of 10-20°.

需要2個照明器。一個是顯微鏡内部的一個反射式亮場(chǎng)照明器，另一個是一個可調(diào)的外部可聚焦輔助照明器，以提供10-20°角的反射斜照明。

The filter assembly for retaining particulate contamination consists of a filter holder made of glass or other suitable material, and is equipped with a vacuum source and a suitable membrane filter.

用於(yú)保留顆粒污染物的過濾器組件包括由玻璃或其他合适材料制成的過濾器支架，並(bìng)配備有真空源和合适的膜過濾器。

The membrane filter is of suitable size, black or dark grey in colour, non-gridded or gridded, and 1.0 µm or finer in nominal pore size.

膜過(guò)濾器尺寸合适
，顔色爲黑色或深灰色，無網格或網格，标稱(chēng)孔徑爲1.0 µm或更小。

General precautions 一般注意事項

The test is carried out under conditions limiting particulate contamination, preferably in a laminar-flow cabinet.

Very carefully wash the glassware and filter assembly used, except for the membrane filter, with a warm detergent solution and rinse with abundant amounts of water to remove all traces of detergent. Immediately before use, rinse both sides of the membrane filter and the equipment from top to bottom, outside and then inside, with particle-free water R.

試驗在限制顆粒污染的條件下進行
，最好在層(céng)流櫃(guì)中進行。

使用溫熱的清潔劑溶液仔細清洗所用的玻璃器皿和過濾器組件（膜式過濾器除外），並(bìng)用大量水沖洗，以去除所有清潔劑痕迹。使用前，用無顆粒水從上到下、從外到内沖洗膜過濾器和設備(bèi)的兩側。

In order to check that the environment is suitable for the test, that the glassware and the membrane filter are properly cleaned and that the water to be used is particle-free, the following test is carried out: determine the particulate contamination of a 50 mL volume of particle-free water R according to the method described below. If more than 20 particles 10 µm or larger in size or if more than 5 particles 25 µm or larger in size are present within the filtration area, the precautions taken for the test are not sufficient. The preparatory steps must be repeated until the environment, glassware, membrane filter and water are suitable for the test.

爲瞭(le)檢查環境是否适合試驗，玻璃器皿和膜過濾器是否正確(què)清潔，所使用的水是否無顆粒，應進行以下試驗：根據下述方法測定50 mL無顆(kē)粒水的顆(kē)粒污染情況。如果過(guò)濾區域内存在20個以上粒徑爲10 µm或更大的顆粒，或5個以上粒徑爲25 µm或更大的顆粒，則試驗所採(cǎi)取的預防措施是不夠的。必須重複準備(bèi)步驟，直到環境、玻璃器皿、膜過濾器和水适合試驗。

Method方法

Mix the contents of the samples by slowly inverting the container 20 times successively. If necessary, cautiously remove the sealing closure. Clean the outer surfaces of the container opening using a jet of particle-free water R and remove the closure, avoiding any contamination of the contents.

通過将容器連續緩慢翻轉20次來混合樣品内容物。如有必要，小心地拆下密封蓋(gài)。使用無顆粒水水流清潔容器開口的外表面，並(bìng)拆下封蓋(gài)，避免内容物的任何污染
。

For large-volume parenterals, single units are tested. For small-volume parenterals less than 25 mL in volume, the contents of 10 or more units are combined in a cleaned container; where justified and authorised, the test solution may be prepared by mixing the contents of a suitable number of vials and diluting to 25 mL with particle-free water R or with an appropriate solvent without contamination of particles when particle-free water R is not suitable. Small-volume parenterals having a volume of 25 mL or more may be tested individually.

對於(yú)大容量腸外注射，需要測(cè)試單個樣品。對於(yú)體積小於(yú)25 mL的小容量腸外注射劑，将10個或更多個的樣品組合在一個清潔的容器中；在合理和授權的情況下，可通過混合适當(dāng)數量的小瓶中的内容物，並(bìng)用無顆粒水稀釋至25 mL，當(dāng)無顆粒水不适用時，可用無顆粒污染的适當(dāng)溶劑(jì)稀釋至25 mL來制備試驗溶液。體積爲25 mL或以上的小容量非腸(cháng)道藥物可單(dān)獨進行試驗
。

Powders for parenteral administration are constituted with particle-free water R or with an appropriate solvent without contamination of particles when particle-free water R is not suitable.

用於(yú)腸(cháng)外給藥的粉末由無顆粒水配置，或當無顆粒水不适用時，使用不含污染顆粒的溶劑重新配制。

The number of test specimens must be adequate to provide a statistically sound assessment. For large-volume parenterals or for small-volume parenterals having a volume of 25 mL or more, fewer than 10 units may be tested, based on an appropriate sampling plan.

試樣數量必須足以提供統計上合理的評估。對於(yú)大容量腸(cháng)外注射劑或體積大於(yú)等於(yú)25 mL的小容量腸外注射劑，根據适當的取樣計劃，可測(cè)試少於(yú)10個樣品。

Wet the inside of the filter holder fitted with the membrane filter with several millilitres of particle-free water R. Transfer to the filtration funnel the total volume of a solution pool or of a single unit, and apply vacuum. If needed, add stepwise a portion of the solution until the entire volume is filtered. After the last addition of solution, begin rinsing the inner walls of the filter holder by using a jet of particle-free water R. Maintain the vacuum until the surface of the membrane filter is free from liquid. Place the filter in a Petri dish and allow the filter to air-dry with the cover slightly ajar. After the filter has been dried, place the Petri dish on the stage of the microscope, scan the entire membrane filter under the reflected light from the illuminating device, and count the number of particles that are equal to or greater than 10 µm and the number of particles that are equal to or greater than 25 µm. Alternatively, partial filter count and determination of the total filter count by calculation is allowed. Calculate the mean number of particles for the preparation to be examined.

用幾毫升無顆粒水濕潤裝有薄膜過濾器的過濾器支架内部。将溶液或單個樣品轉移到過濾漏鬥中，並(bìng)施加真空。如果需要，逐步添加一部分溶液，直到整個體積被過濾。最後一次添加溶液後，使用無顆粒水水流開始沖洗過濾器支架的内壁。保持真空，直到過濾膜表面沒有液體。将過濾膜放置在有蓋培養皿中，讓過濾膜風幹，蓋子稍微半開。過濾膜幹燥後，将皮氏培養皿放在顯微鏡台上，在照明設備的反射光下掃描整個膜過濾器，並(bìng)計算等於(yú)或大於(yú)10 µm的顆粒數量和等於或大於25 µm的顆粒數量。或者，如果計算方式允許，可以對(duì)部分濾膜上的顆粒計數，並(bìng)決定總濾膜上的顆粒數量。計算待檢查制劑的平均顆粒數。

The particle sizing process with the use of the circular diameter graticule is carried out by transforming mentally the image of each particle into a circle and then comparing it to the 10 µm and 25 µm graticule reference circles. Thereby the particles are not moved from their initial locations within the graticule field of view and are not superimposed on the reference circles for comparison. The inner diameter of the transparent graticule reference circles is used to size white and transparent particles, while dark particles are sized by using the outer diameter of the black opaque graticule reference circles.

通過(guò)将每個(gè)顆粒的圖像轉換成一個(gè)圓，然後将其與10 µm和25 µm的分劃參考圓進行比較，使用圓形直徑分劃的顆粒尺寸來執行顆粒尺寸測定過程。因此，粒子不會從網格視場内的初始位置移動，也不會疊加在參考圓上進行比較。透明分劃參考圓的内徑用於(yú)確(què)定白色和透明粒子的大小，而黑色粒子的大小則使用黑色不透明分劃參考圓的外徑進行確(què)定。

In performing the microscopic particle count test do not attempt to size or enumerate amorphous, semi-liquid, or otherwise morphologically indistinct materials that have the appearance of a stain or discoloration on the membrane filter. These materials show little or no surface relief and present a gelatinous or film-like appearance. In such cases the interpretation of enumeration may be aided by testing a sample of the solution by the light obscuration particle count test.

在進行微觀顆粒計數試驗時，不要試圖測(cè)量或列舉無定形、半液體或其他形态不明顯的材料，這些材料在膜過濾器上出現污漬或變(biàn)色。這些材料很少或沒有表面浮雕，呈現膠狀或薄膜狀外觀。在這種情況下，可通過光阻法粒子計數測(cè)試溶液樣品來輔助解釋計數。

Evaluation評估

For preparations supplied in containers with a nominal volume of more than 100 mL, apply the criteria of test 2.A.

For preparations supplied in containers with a nominal volume of less than 100 mL, apply the criteria of test 2.B.

♦For preparations supplied in containers with a nominal volume of 100 mL, apply the criteria of test 2B.♦

If the average number of particles exceeds the limits, test the preparation by the microscopic particle count test.

對於在标稱體積大於100 mL的容器中提供的制劑(jì)，應採(cǎi)用試驗2 A.的标準。

對於在标稱體積小於100 mL的容器中提供的制劑(jì)，採(cǎi)用試驗2 B的标準。

♦對於在标稱體積爲100 mL的容器中提供的制劑(jì)，採(cǎi)用試驗2 B的标準。♦

如果平均顆粒數超過限值，則通過顯微顆粒計數測(cè)試對(duì)制劑進行測(cè)試。

Test 2.A – Solutions for infusion or solutions for injection supplied in containers with a nominal content of more than 100 mL.

The preparation complies with the test if the average number of particles present in the units tested does not exceed 12 per millilitre equal to or greater than 10 µm and does not exceed 2 per millilitre equal to or greater than 25 µm.

試驗2A–輸液溶液或注射溶液，裝在标稱(chēng)含量超過(guò)100 mL的容器中。

如果被測(cè)單位中顆粒尺寸等於(yú)或大於(yú)10 µm的平均數量不超過每毫升12個，且顆粒尺寸等於或大於25 µm的平均數量不超過每毫升2個，則制劑符合試驗要求。

Test 2.B – Solutions for infusion or solutions for injection supplied in containers with a nominal content of less than 100 mL

The preparation complies with the test if the average number of particles present in the units tested does not exceed 3000 per container equal to or greater than 10 µm and does not exceed 300 per container equal to or greater than 25 µm.

試驗2B–在标稱含量小於100 mL的容器中提供的輸(shū)液溶液或注射溶液。

如果被測(cè)單元中存在的尺寸等於(yú)或大於(yú)10 µm的顆粒平均數不超過3000個/容器，且尺寸大於等於25 um的顆粒平均數不超過300個/容器，則制劑(jì)符合試(shì)驗要求。

檢(jiǎn)測(cè)結果判定标準總結：