洋葱牛肉卷1. Purpo: To establish a method for microbial limit test, and to ensure the smooth
progress of microbial limit test.
2. Scope of Application: This procedure is applicable for the microbial limit test of
health food.
前合后仰3. Responsibilities: The QC laboratorian are responsible for the implementation of
this procedure and the laboratory director shall assume the
responsibility of supervision and inspection for the effective
implementation of this procedure.
4. Contents:
美容常识4.1 On the basis of the new version of GB4789.5 -2010/2012
4.2 Brief description:
Microbial limit test method refers to a method for detecting the degree of contamination to which the non-specified sterile preparation, raw materials, excipients are subject, including the test for the amount of bacterium contamination, as well as control bacteria and pathogenic bacteria.
The test products shall be randomly sampled. Sampling amount is usually three times of testing amount (more than two minimum packing unit). The whole process of inspection shall strictly comply with aptic operations to prevent recontamination.
Unless otherwi specified, the incubation temperature of bacteria and control bacteria is 30 ~ 3 5 °C in this test method, and that of fungi and yeast is 25 ~
28 °C.
The report for the test results takes 1 g, 1 ml or 10cm2 as the unit.
4.3 Determination of total amount of bacteria and mold
4.3.1 Brief description:
Count of bacteria, fungi, yeasts is determining the number of viable bacteria growing in the non-sterile preparations within specified business units, which is an important indicator for judging the degree of microbial contamination in the product, Also, it is one of the comprehensive bas for hygiene evaluation of products, raw materials, excipients, equipments and appliances, process, production environment and the health status of the operator's in the manufacturing enterpri.
Plate count method, which is one of viable bacteria count methods, is ud for detecting bacteria, fungi and yeasts, and is currently ud in many countries.
This method takes the visible and individual colonies formed on the agar plates by each bacterium (Nutrient Agar), fungi (Ro Bengal Agar), yeasts (Ro Bengal Agar or Yeast Peptone Dextro Agar) as the counting basis. The measured results reflect the colonies number of bacteria (a group of mesophilic, aerobic and facultative anaerobic bacteria grown on Nutrient Agar), fungi and yeast growing under specified conditions. Bacteria, fungi and yeasts that have specific requirements for nutrients, oxygen, temperature, PH and other factors are excluded.
4.3.2 Equipments, instruments and appliances:
4.3.2.1 Sterile room:
4.3.2.1.1 Sterile room shall be with good natural lighting, prevented from dampness,
and away from the toilets and contaminated area, composing of buffer room
(2 rooms) and operating room.
There shall be a sample transferring box between the operating room and the
buffering room.
Six surfaces of the sterile room shall be smooth and can withstand cleaning
and disinfection. The connecting joints between walls and floors, and that
between walls and ceiling shall be concave-curved, amless, with no dead
corner. Sewer shall not be installed within the operating room.
4.3.2.1.2 Operating room: operating room shall be installed with clean bench, of which
the clean class is 100,000, and partial is 100.
4.3.2.1.3 Buffer room: washing basin, sterile clothing, hats, masks, slippers and so
on shall be placed in the buffer room. Incubators and other sundries shall not
be placed within a buffer room.
4.3.2.2 Other equipments: clean bench, constant temperature incubator, shaker,
electrothermal blowing dry box, refrigerator, autoclave.
4.3.2.3 Instruments and appliances: Drug balance, conical flasks, culture dish, pipette,
微信发不了图片怎么回事
test tubes, sterile sampling paper, scissors, tweezers, medicine spoon, cotton with ethanol, sterile overalls.
4.3.3 Diluent and its preparation:
4.3.3.1 0.9% sterile sodium chloride solution: take 9.0g of sodium chloride and dilute
with water to 1000 ml for dissolution, distribute into conical flasks, sterilize at 121 °C for 20 minutes for u.
4.3.3.2 Sterile phosphate buffer solution (pH7.2): take 2自编故事
5.8g of sodium hydrogen
phosphate and 4.4g of sodium dihydrogen phosphate, dilute with water to 1000 ml, distribute into conical flasks, sterilize at 121°C for 20 minutes for u.
4.3.4 Medium: Nutrient Agar medium, Ro Bengal Agar Medium.
4.3.5 Sampling and testing amount of test products
4.3.
5.1 Sampling:
4.3.
5.1.1 Generally random sampling method is ud, sampling size is usually
three times of testing amount which is more than two minimum packing unit
(to prepare for re-examination).
4.3.
5.1.2 When sampling, in ca of abnormal or suspicious samples, suspicious
samples shall be lected. The package with obvious breakage can not be
taken as sample.
4.3.
5.1.3 Products with mites, fungi, worms which can be en from the appearance
of products and bottle (the inner side of the lid and neck) and deterioration
are directly judged as unqualified products. And there is no need for
sampling.
4.3.
5.2 Test amount
4.3.
5.2.1 Test amount of all dosage forms are required of more than two packing units.
4.3.
5.2.2 Test amount of solid and mi-solid (viscous test products) preparation
is 10g.
4.3.
5.2.3 Test volume of liquid preparation is 10ml.
4.3.6 Method of operation:
4.3.6.1 Preparation before operation
4.3.6.1.1 The test products and all the sterile dishes, conical flasks, test tubes,
pipette (1ml, 10ml), measuring cylinder, diluent are moved to a sterile
room. All the items ud in each test must be prepared in advance. Adequate
amount is needed to avoid the in and out of the operating room. All outside
packages (kraft) are removed after coded.
4.3.6.1.2 Open UV lamp and clean bench in the sterile room, and make it work for no
less than 30 min.
4.3.6.1.3 The operator washes hands with soap and clos UV lamp before into the
buffer room, change working shoes after going into the buffer
room. Then wash hands with 0.1% benzalkonium bromide solution or other
disinfectant or swab with cotton containing ethanol, wear sterile clothing, hats,
masks and gloves.
4.3.6.1.4 Swab hands with cotton containing ethanol before operation at first, then
swab the opening of the test product bottles, boxes, bags with cotton
containing ethanol, and unal the test products with sterile scissors after the
alcohol volatilized.
4.3.6.2 Preparation of test solution:
4.3.6.2.1 Liquid test products: take 10ml of test products, add into 90ml of sterile
sodium chloride-peptone buffer (pH7.0), mix well, as the test solution (1:10).
4.3.6.2.2 Solid, mi-solid or viscous liquid test products: weigh 10g of the test
products, add into 100ml of sterile sodium
chloride-peptone buffer(pH7.0 ), heat to dissolve in a constant temperature
water bath, but the temperature shall not exceed 45 °C, and then shake on
the shaker, mix well, as the test solution (1:10).
4.3.6.2.3 Test products of enteric tablet: weigh 10 g of the test products, add into flask
with 100 ml of sterilized phosphate buffer (pH6.8), heat in water bath at 45 °C,
shake to dissolve, as the test solution (1:10).
4.3.6.3 Dilution of the test solution (diluted with increments of 10 times)
4.3.6.3.1 Take two small sterile test tubes, add sterile sodium chloride-peptone buffer
(pH7.0) with a pipette, plug the tube immediately after addition of buffer.
4.3.6.3.2 Take another 1ml sterile pipette to add 1ml of homogeneous test solution
(1:10) into a test tube with 9ml of sterilized sodium chloride-peptone buffer
tube (pH7.0), mis well, namely the test solution (1:100), and so on.
Each dilution must change a pipette.
When diluted with increments of 10 times, dip the pipette into the dilution
solution (level 1) not lower than 2.5cm from liquid surface, repeated suck and
blow for about 10 times, the liquid level shall be a little above the upper scale大学生抗疫
of the pipette when suction, then lift the pipette and stick to the tube wall to
adjust the liquid level to the mark, then slowly blow out all the test solution
along the inner wall of the dilution tube (level 2) near the liquid surface (no
touch to the liquid) (no sticking or residual liquid shall be left in the pipette). 4.3.6.4 While diluted with increments of 10 times in dishes, suck 1ml of diluted solution
of various levels into each sterile dish with the corresponding pipette (when
dilute from high level to low level the same pipette can be ud during the
suction). 2 ~ 3 dishes are prepared in each dilution level (in this ca, usually
dish in the left hand with the lid half open, pipette in the right hand). During the祝你生日快乐英文
pouring, 1 ml of the test solution will be slowly poured, no residual liquid left in
the tube, to prevent anti-flowing into the tip of the pipette. At the same time as
花与蛇1a negative control (after the completion of pouring each level of dilution
fluid, suck 1ml of diluent of each level with 1 ml pipette into four dishes,
respectively, in which two as negative controls for the number of bacteria and
