2.9.19. PARTICULATE CONTAMINATION: SUB-VISIBLE PARTICLES

2.9.19. 顆粒物污染：亞可見顆粒物檢查規(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.

對于顆粒物污染的測定包含兩種方法，下文進(jìn)行了詳細(xì)介紹：方法1（光阻法顆粒計數(shù)試驗）和方法2（顯微顆粒物計數(shù)試驗）。當(dāng)檢查注射劑和輸液中是否存在亞可見顆粒時，最好使用方法1。然而，可能有必要通過光阻法顆粒計數(shù)試驗和顯微顆粒計數(shù)試驗來測試某些制劑，以得出符合要求的結(jié)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.

并非所有非腸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 不適用時，應(yīng)根據(jù)方法2進(jìn)行測試，如乳液、膠體和脂質(zhì)體制劑等。 同樣，在吸入傳感器時會產(chǎn)生空氣或氣泡的產(chǎn)品也可能需要進(jìn)行顯微顆粒物計數(shù)測試。 如果待測制劑的粘度足夠高以至于無法通過任一測試方法對其進(jìn)行檢查，則可以根據(jù)需要，用適當(dāng)?shù)南♂寗┻M(jìn)行定量稀釋以降低粘度，以便進(jìn)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.

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

METHOD 1. LIGHT OBSCURATION PARTICLE COUNTTEST

光阻法顆粒計數(shù)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(jù)光阻原理，使用合適的儀器，可以根據(jù)大小自動確定顆粒大小和顆粒數(shù)量。使用合適的認(rèn)證標(biāo)準(zhǔn)物質(zhì)對儀器進(jìn)行校準(zhǔn)，該標(biāo)準(zhǔn)物質(zhì)由已知尺寸在10 µm到25 µm之間的球形顆粒的分散體組成。這些標(biāo)準(zhǔn)顆粒分散在無顆粒水中。分散過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(jìn)行，最好在層流柜中進(jìn)行。使用溫?zé)岬那鍧崉┤芤鹤屑?xì)清洗所用的玻璃器皿和過濾設(shè)備（膜式過濾器除外），并用大量水沖洗，以去除所有清潔劑痕跡。使用前，用無顆粒水從上到下、從外到內(nèi)沖洗設(shè)備。

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.

注意不要將氣泡引入待檢查的制劑中，尤其是將制劑的部分轉(zhuǎn)移到要進(jìn)行測定的容器中時。

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。

為了檢查環(huán)境是否適合試驗，玻璃器皿是否正確清潔，所使用的水是否無顆粒，應(yīng)進(jìn)行以下試驗：根據(jù)下述方法，測定5個無顆粒水樣品的顆粒污染情況，每個樣品5 mL。如果10 µm或更大粒徑的顆粒數(shù)量超過25個，則為試驗采取的預(yù)防措施是不夠的。必須重復(fù)準(zhǔn)備步驟，直到環(huán)境、玻璃器皿和水適合試驗

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(xù)20次緩慢翻轉(zhuǎn)容器，混合樣品里的內(nèi)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，并拆下封蓋，避免內(nèi)容物受到任何污染。通過適當(dāng)?shù)拇胧┫龤馀荩珈o置2分鐘或超聲波處理。

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.

對于大容量腸外注射劑，需要測試單個單元。對于體積小于25 mL的小容量腸外注射劑，將10個或更多個的內(nèi)容物組合在一個清潔的容器中，以獲得不小于25 mL的體積的溶液；在合理和授權(quán)的情況下，可通過混合適當(dāng)數(shù)量的小瓶中的內(nèi)容物，并用無顆粒水稀釋至25 mL，或在無顆粒水不適用時，用無顆粒污染的適當(dāng)溶劑稀釋至25 mL來制備試驗溶液。體積為25 mL或以上的小容量非腸道藥物試劑可單獨進(jìn)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.

用于腸外給藥的粉末用無顆粒水重新配制，或當(dāng)無顆粒水不適用時，使用適當(dāng)?shù)臒o顆粒污染的溶劑重新進(jìn)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。

測試樣本的數(shù)量必須足夠，以提供統(tǒng)計上合理的評估。根據(jù)適當(dāng)?shù)某闃佑媱潱瑢τ诖笕萘炕?/span>25 mL以上的小容量腸外靜脈，可檢測小于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，每份不少于5 mL，并計算等于或大于10 µm和25 µm的顆粒數(shù)。忽略第一份的結(jié)果，并計算待檢查制劑的平均顆粒數(shù)。

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.

對于在標(biāo)稱體積超過100 mL的容器中提供的制劑，應(yīng)采用試驗1A的標(biāo)準(zhǔn)。

對于在標(biāo)稱體積小于100 mL的容器中提供的制劑，采用試驗1B的標(biāo)準(zhǔn)。

對于在標(biāo)稱體積為100 mL的容器中提供的制劑，采用試驗1B的標(biāo)準(zhǔn)。

如果平均顆粒數(shù)超過限值，則通過顯微顆粒計數(shù)測試對制劑進(jìn)行測試。

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，裝在標(biāo)稱含量超過100 mL的容器中。

如果被測單位樣品中粒徑等于或大于10 µm的顆粒的平均數(shù)量不超過每毫升25個，且粒徑等于或大于25 µm的顆粒平均數(shù)量不超過每毫升3個，則制劑符合試驗要求。

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，裝在標(biāo)稱含量小于100 mL的容器中。

如果被測單元中粒徑等于或大于10 µm的顆粒的平均數(shù)量不超過6000個/容器，且粒徑等于或大于25 µm的顆粒平均數(shù)量不超過600個/容器，則制劑符合試驗要求。

METHOD 2. MICROSCOPIC PARTICLE COUNT TEST

顯微顆粒計數(shù)

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

使用合適的雙目顯微鏡、過濾器組件（用于保留顆粒污染物）和薄膜過濾器（用于檢查）。

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微計（使用物鏡測微計校準(zhǔn)）、一個機械工作臺（能夠容納并穿過膜過濾器的整個過濾區(qū)域）、2個合適的照明器（除了傾斜照明外，還提供反射照明），并將其調(diào)整為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).

目鏡測微計是一個圓形直徑分劃器（見圖2.9.19.-1）由一個被十字準(zhǔn)線分成四個象限的大圓、100倍放大時直徑為10 µm和25 µm的透明和黑色參考圓、以及以10 µm增量刻度的線性刻度組成。使用經(jīng)過國內(nèi)或國際標(biāo)準(zhǔn)機構(gòu)認(rèn)證的測微計對其進(jìn)行校準(zhǔn)。分劃線性刻度的相對誤差在±2%以內(nèi)是可以接受的。大圓被designated為網(wǎng)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。一個是顯微鏡內(nèi)部的一個反射式亮場照明器，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.

用于保留顆粒污染物的過濾器組件包括由玻璃或其他合適材料制成的過濾器支架，并配備有真空源和合適的膜過濾器。

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.

膜過濾器尺寸合適，顏色為黑色或深灰色，無網(wǎng)格或網(wǎng)格，標(biāo)稱孔徑為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(jìn)行，最好在層流柜中進(jìn)行。

使用溫?zé)岬那鍧崉┤芤鹤屑?xì)清洗所用的玻璃器皿和過濾器組件（膜式過濾器除外），并用大量水沖洗，以去除所有清潔劑痕跡。使用前，用無顆粒水從上到下、從外到內(nèi)沖洗膜過濾器和設(shè)備的兩側(cè)。

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.

為了檢查環(huán)境是否適合試驗，玻璃器皿和膜過濾器是否正確清潔，所使用的水是否無顆粒，應(yīng)進(jìn)行以下試驗：根據(jù)下述方法測定50 mL無顆粒水的顆粒污染情況。如果過濾區(qū)域內(nèi)存在20個以上粒徑為10 µm或更大的顆粒，或5個以上粒徑為25 µm或更大的顆粒，則試驗所采取的預(yù)防措施是不夠的。必須重復(fù)準(zhǔn)備步驟，直到環(huán)境、玻璃器皿、膜過濾器和水適合試驗。

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.

通過將容器連續(xù)緩慢翻轉(zhuǎn)20次來混合樣品內(nèi)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，并拆下封蓋，避免內(nèi)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.

對于大容量腸外注射，需要測試單個樣品。對于體積小于25 mL的小容量腸外注射劑，將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(quán)的情況下，可通過混合適當(dāng)數(shù)量的小瓶中的內(nèi)容物，并用無顆粒水稀釋至25 mL，當(dāng)無顆粒水不適用時，可用無顆粒污染的適當(dāng)溶劑稀釋至25 mL來制備試驗溶液。體積為25 mL或以上的小容量非腸道藥物可單獨進(jìn)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.

用于腸外給藥的粉末由無顆粒水配置，或當(dāng)無顆粒水不適用時，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(shù)量必須足以提供統(tǒng)計上合理的評估。對于大容量腸外注射劑或體積大于等于25 mL的小容量腸外注射劑，根據(jù)適當(dāng)?shù)娜佑媱潱蓽y試少于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內(nèi)部。將溶液或單個樣品轉(zhuǎn)移到過濾漏斗中，并施加真空。如果需要，逐步添加一部分溶液，直到整個體積被過濾。最后一次添加溶液后，使用無顆粒水水流開始沖洗過濾器支架的內(nèi)壁。保持真空，直到過濾膜表面沒有液體。將過濾膜放置在有蓋培養(yǎng)皿中，讓過濾膜風(fēng)干，蓋子稍微半開。過濾膜干燥后，將皮氏培養(yǎng)皿放在顯微鏡臺上，在照明設(shè)備的反射光下掃描整個膜過濾器，并計算等于或大于10 µm的顆粒數(shù)量和等于或大于25 µm的顆粒數(shù)量。或者，如果計算方式允許，可以對部分濾膜上的顆粒計數(shù)，并決定總濾膜上的顆粒數(shù)量。計算待檢查制劑的平均顆粒數(shù)。

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.

通過將每個顆粒的圖像轉(zhuǎn)換成一個圓，然后將其與10 µm和25 µm的分劃參考圓進(jìn)行比較，使用圓形直徑分劃的顆粒尺寸來執(zhí)行顆粒尺寸測定過程。因此，粒子不會從網(wǎng)格視場內(nèi)的初始位置移動，也不會疊加在參考圓上進(jìn)行比較。透明分劃參考圓的內(nèi)徑用于確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(jìn)行確定。

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

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.

對于在標(biāo)稱體積大于100 mL的容器中提供的制劑，應(yīng)采用試驗2 A.的標(biāo)準(zhǔn)。

對于在標(biāo)稱體積小于100 mL的容器中提供的制劑，采用試驗2 B的標(biāo)準(zhǔn)。

?對于在標(biāo)稱體積為100 mL的容器中提供的制劑，采用試驗2 B的標(biāo)準(zhǔn)。?

如果平均顆粒數(shù)超過限值，則通過顯微顆粒計數(shù)測試對制劑進(jìn)行測試。

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，裝在標(biāo)稱含量超過100 mL的容器中。

如果被測單位中顆粒尺寸等于或大于10 µm的平均數(shù)量不超過每毫升12個，且顆粒尺寸等于或大于25 µm的平均數(shù)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–在標(biāo)稱含量小于100 mL的容器中提供的輸液溶液或注射溶液。

如果被測單元中存在的尺寸等于或大于10 µm的顆粒平均數(shù)不超過3000個/容器，且尺寸大于等于25 um的顆粒平均數(shù)不超過300個/容器，則制劑符合試驗要求。

檢測結(jié)果判定標(biāo)準(zhǔn)總結(jié)：