City water, Hospital, Pharmaceutical, Food, Medical device, Injections

Biologicals, Cellular therapies, Tissue culture, Biotechnology Products  

Aseptic process treatment, monitoring and validation of water, liquids, raw materials, products are very important at hospital, pharmaceutical, Injections, food, beverage, medical device, biologicals, tissue culture, blood and cellular therapies products production and application

1. Fast, effective aseptic monitoring & validation for clear correct aseptic

     process treatment and products production & application

2. Fast, effective aseptic purification design and application for clear, fast,

    effective aseptic process treatment and products production  

1. The conventional aseptic process treatment design and application (except RO

    and distiller unit) are hard to purify and separate the bacteria, virus, pyrogenic

     bacteria, inactive organisms, endotoxin, pyrogen clear to do the aseptic water, 

     liquid, raw materials and products effectively

The aseptic process treatment of water, liquid, raw material and products are not so easy to kill, purify and separate the bacteria, virus, pyrogenic organisms, endotoxin, pyrogen clear effectively up to bacteria-free, germ-free, endotoxin-free and pyrogen-free by the conventional chemical disinfectant, UV, heat, filter treatment (except distiller and RO unit)

The standard aseptic process treatment to sterilize, purify and separate the bacteria, virus, pyrogenic organisms, endotoxin, pyrogen clear effectively up to bacteria-free, germ-free, endotoxin-free and pyrogen-free are divided into the different kind and type of aseptic sterilization and purification design and application:  

1. The standard aseptic sterilization process treatment

    apply the clear expected sterilization temperature, dosage and functional time to destroy

    the bacteria, virus, fungi and pyrogenic organisms into inactive carcass, also break the

    endotoxin and pyrogen into dissolution and dispersion clear completely up to bacteria-

free, germ-free, endotoxin-free and pyrogen-free effectively

    a. dry heat: 250 ^oC, 2~4 hours,     b. steam: 131 ^oC, 2~4 hours  

2. The standard aseptic purification process treatment

     apply the aseptic finer membrane filter which smaller than the particles size of bacteria,

virus, fungi, pyrogenic organisms, endotoxin and pyrogen to purify and remove the

     particles of bacteria, virus, fungi, pyrogenic organisms, endotoxin and pyrogen clear

     completely up to bacteria-free, germ-free, endotoxin-free and pyrogen-free effectively

    a. the particle size of bacteria, fungi, pyrogenic organism at the range of 0.22~20 micron

    b. the particle size of virus, endotoxin, pyrogen at the range of 0.1~0.001 micron

  1) aseptic 0.45~0.2 micron membrane filter can be purified the bacteria, fungi, pyrogenic

      organisms clear effectively, but low effect to purify virus, endotoxin, pyrogen clear

  2) aseptic UF/NF/RO membrane filter can be purified and removed the bacteria, fungi, 

       pyrogenic organisms, virus, endotoxin, pyrogen clear effectively and completely       

Most of customers are normally considered that the water, liquid, raw materials, products would be bacteria free after conventional chemical disinfectant, UV, heat aseptic treatment, Eventually, the conventional aseptic treatment are only applied to inhibit the bacteria, virus,  fungi, and pyrogenic organisms into inactive organisms and lose the activity temporarily; however, when the inactive organisms are compatible to the inhibitive environment, the inactive organisms would be growth up into the further bacterial contaminations, Except the   conventional aseptic treatment are established the standard aseptic treatment to do the clear, correct aseptic sterilization and disinfection process treatment

As to aseptic purification process treatment, Most of customers are normally considered that the water, liquid would be bacteria free and pyrogen free after conventional MF/SMF/ UF/NF/RO membrane filter filtration treatment are incorrectly, Due to the conventional membrane filter are microorganisms breeding bed, If the conventional membrane filters are aseptic, the purified water and liquid should be bacteria free and pyrogen free, If the filters are not aseptic, the purified water and liquid should be contaminated much contents of bacterial contaminations after filtration  

2. The conventional aseptic monitoring and validation are hard to monitor and

validate the bacteria, virus, pyrogenic bacteria, inactive organisms, endotoxin,

pyrogen to be purified clear fastly and effectively or not, for clear correct

     aseptic process treatment, aseptic products production and application  

The aseptic monitoring and validation of bacteria, virus, pyrogenic organisms, endotoxin, pyrogen monitoring and sterilization validation are not so simple, easy to monitor and validate that the aseptic water, liquid, raw material and product are bacteria free, germ free, endotoxin free and pyrogen free or not, which must be performed the following step and procedure to monitor and validate that the aseptic water, liquid, raw material and product are really bacteria free, germ free, endotoxin free and pyrogen free or not,  

1. first step to monitor and validate the growth promotion test of culture media and 

    microbiological sensitivity test  

2. secondary step to monitor and validate the residues and contents of bacteria, virus, pyrogenic 

    bacteria, endotoxin, pyrogen test  

The conventional aseptic monitoring and validation have not much beneficial to aseptic treatment and application, Due to the aseptic analysis time are needed much times for test, Normally needed 24 hours test time, after testing, the water, liquids, raw material, products are already used and completely, and the test result and data of aseptic bacteria, virus, pyrogenic organisms, endotoxin, pyrogen monitoring and validation are always incorrectly and only acted as the reference data, due to the aseptic monitoring and validation test effect and result are always influenced by the quality of culture media varied, the inhibition of disinfectant, steriliant  

Fast, correct aseptic treatment monitoring and validation will be new direction and main topics at the future aseptic treatment, monitoring, validation development and application

The contents of bacteria, virus, pyrogenic bacteria, endotoxin and pyrogen of process water are main bacterial contaminations of the aseptic process treatment of hospital, pharmaceutical, injection solutions, food, beverage, medical device, biologicals, cellular, tissue, and blood products, and also be the main point of aseptic process treatment to sterilize, purify and reduce the contents of bacteria, virus, pyrogenic bacteria, endotoxin and pyrogen clear effectively

If the process water can be pre-treated, pre-monitored, pre-validated up to germ free, bacteria free, endotoxin free and pyrogen free water initially, The aseptic process treatment have already been completed 50% opportunity to produce out the aseptic water, liquids, raw materials and products to meet the standard of aseptic monitoring and validation  

1. aseptic process:  to establish the non-bacterial contaminations infectional process to

                               sterilize, purify and separate the bacteria, virus, pyrogenic bacteria,

                               endotoxin, pyrogen clear up to bacteria-free, germ-free, endotoxin

                               free, pyrogen free water, liquids, raw materials, and products

2. aseptic treatment: to establish the high efficiency sterilization, purification design to

                                sterilize, purify and separate the bacteria, virus, pyrogenic bacteria,

                                endotoxin, pyrogen clear completely to do bacteria-free, germ-free,

                                endotoxin free, pyrogen free water, liquids, raw materials, products

3. aseptic monitoring & validation:   to establish the clear, correct microbiological and

                                sterilization validation test to monitor and validate the residues and

                                contents of aseptic water, liquids, raw materials, products to meet

                                the standard of aseptic monitoring and validation or not  

The conventional aseptic process treatment of hospital, pharmaceutical, food, beverage, medical device, biologicals, cellular products are normally applied the high dosage of chemical inhibitor, disinfectant of formalin, methanol, NaClO, ClO₂ to insure the aseptic water, liquids, raw materials, products up to bacteria free effectively; however, The aseptic treatment of high dosage of chemical inhibitor, disinfectant of NaClO, ClO2, formalin, methanol are also transformed the aseptic water, liquids, raw materials, products into toxic water, liquids, raw materials, products for application

like: the draught beer add the formalin to be bacteria free, The city water add 0.8~1.0 ppm

         NaClO,ClO₂ to prevent the SARS, enterovirus being contaminations again

The aseptic monitoring and validation of aseptic water, liquids, raw materials, products are not more the safeguard of sanitary and safety, some of economic developed country have already performed the toxicity test for insuring the aseptic water, liquids, raw materials, products to be safety, sanitary, toxic free

Toxicity test would be replaced the aseptic test to safeguard the aseptic water, liquids, raw materials, products to be safety, sanitary, toxic free  

The conventional sterilization monitoring, validation of pharmaceutical, food, beverage, hospital, medical device, biologicals, cellular products are normally applied the steam, dry heat, EO-gas, radiation sterilization treatment to destroy, disintegrate the active organisms into the inactive organisms, carcass up to aseptic water, liquids, raw materials, products

1. disintegration sterilization: a. steam: 131^oC, 2~4 hours    b. dry heat: 250 ^oC, 2~4 hours

                                                c. before sterilization bacteria count: 100 CFU/ml

                                                d. after sterilization pyrogen residues: ≦0.2 EU

2. standard sterilization:   a. steam: 121^oC, 30 minutes    b. dry heat: 160 ^oC, 30~45 minutes

                                          c. before sterilization bacteria count: 100 CFU/ml

                                          d. after sterilization bacteria count: 10^-6 CFU/ml

3. disinfection sterilization:   a. steam: 111^oC, 135 min.  b. dry heat: 149 ^oC, 45~60min.

                                               c. before sterilization bacteria count: 100 CFU/ml

                                               d. after sterilization bacteria count: 10^-6 CFU/ml

[effect] The sterilization effect of different sterilization temperature, dosage and

              time can be evaluated by the standard sterilization formula:

              sterilization formula:   a.   F t  =  D t  x  (Log A — LogB)

                                               b.   F t  =  F o / L  

     D value: the lethal time up to 90% ate specific temperature, dosage, concentration

     F vaule: the sterilization time after specific temperature, dosage, concentration

     A value: the bacteria count before sterilization

     B value: the bacteria count after sterilization

1. conventional sterilization treatment application:

     most of customers are considered that the application of conventional steam, dry heat, EO-gas, radiation sterilization treatment should be sterilized the sterilized water, liquids, raw materials and products up to bacteria free of aseptic water, liquids, raw materials and products at the long terms and times sterilization, however, they are always found that  the sterilized water, liquids, raw materials and products are still not aseptic at the unclear bacteria contaminations, sterilization temperature, dosage, times being acted on the sterilized water, liquids, raw materials and products

2. conventional sterilization monitoring and validation application:

     most of customers are applied the chemical indicator, 10⁵ CFU/ml of biological indicator

     to monitor and validate that the sterilized water, liquids, raw materials and products are     

     sterilized up to aseptic bacteria free or not, however, most of chemical indicator, 10⁵ CFU

     /ml of biological indicator are only applied to monitor and validate the sterilization      

     temperature, dosage up to effective sterilization temperature, dosage and sterilization rate 

     up to 5D (99.999%) or not

The bacterial count of pre-sterilized water, liquids, raw materials and products are controlled at under 10⁰ CFU/ml, also applied 10⁶ CFU/ml biological indicator at the cooling point of sterilization to monitor and validate the sterilization rate up to 6D will be new aseptic sterilization monitoring, validation direction and application to ensure the residues of aseptic sterilization up to10^-6 CFU/ml

The endotoxin/pyrogenic aseptic treatment, monitoring and validation are best safeguard of aseptic water, liquids, raw materials, products process treatment, monitoring and validation of hospital, pharmaceutical, food, beverage, medical device, biologicals and cellular products, however, due to the conventional de-endotoxin/pyrogen aseptic treatment unit are too expensive and hard to do endotoxin free, pyrogen free of aseptic water, liquids, raw materials, products effectively, and the conventional aseptic monitoring and validation of endotoxin /pyrogen test are still hard to examine the endotoxin/pyrogen contents clear fastly and effectively at the rabbit test (except LAL test) for further effective endotoxin/pyrogen aseptic process treatment application

1. de-endotoxin/pyrogen aseptic process treatment

    a. steam: 131^oC, 2~4 hours, dry heat: 250^oC, 2~4 hours to disintegetrate the endotoxin,

        pyrogen into collapse

     b. aseptic SMF/UF/NF/RO membrane filter to purify the endotoxin/pyrogen contents

        clear effectively and completely

2. aseptic monitoring and validation of endotoxin/pyrogen test

    a. LAL test: to monitor and validate the endotoxin contents of gram-negative bacteria

    b. Rabbit pyrogen test: to monitor, validate the fever response of pyrogens of gram-

                                         negative, gram-positive organisms and other biological pyrogens,

                                         including, yeast, parasitic and virual pyrogens

    c. IPT pyrogen test: to monitor, validate the pyrogens of gram-negative, gram-positive

                                    organisms and other biological pyrogens, including, yeast, parasitic

                                    and virual pyrogens

3. aseptic bacterial residues monitoring and validation of endotoxin/pyrogen test

     a. LAL test: to monitor and validate the endotoxin contents low than 0.2 EU/ml, to be

                        verified the residues of gram-negative organisms low than 10^-6 CFU/ml

     b. IPT pyrogen test: to monitor, validate the pyrogens contents low than 0.2 EU/ml, to be

                        verified the residues of gram-negative, gram-positive organisms and other

                         biological, yeast, parasitic and virual organisms low than 10^-6 CFU/ml

LAL test, IPT pyrogen test as the aseptic endotoxin/pyrogen test, and Will be new innovative aseptic bacterial test to monitor and validate the bacteria residues up to low than 10^-6 CFU/ml

Nano-polymer: instant to concentrate, transform the bacteria, virus, pyrogenic organisms,

                         fungi, endotoxin, pyrogen into the coagulated separative granular silt to

                         purify the bacteria, virus, pyrogenic organisms, fungi, endotoxin, pyrogen

                         clear up to bacteria free, endotoxin free and pyrogen free water

                        a. the bacterial residues can be purified up to≦1~100 CFU/ml or low than

                           ≦10^-2~10^-6 CFU/ml effectively

                        b. the endotoxin/pyrogen residues can be purified up to 0.2 EU effectively  

conventional water aseptic process treatment

1. NaClO, ClO2 of aseptic water treatment

  [point]  apply the low dosage for low bacterial contaminations disinfection treatment

               and high dosage for high bacterial contaminations disinfection treatment

  [defect]  a. after disinfection, the chemical constitutents of NaClO, ClO2 are standing

                    in water which are becomed into the extra chemical toxic contaminants

                b. after disinfection, the aseptic inactive organisms are still standing in water

                    and hard to reduce the pyrogen/endotoxin contens effectively

                c. after disinfection, the bacteria, virus, fungi, pyrogenic organisms are only

                    inhibited to lose the active reactivity temporarily, when it’s compatible with

                    the inhibited environment, it will be growth up being formed the serious

                    bacterial contaminations again

2. MF/SMF/UF/NF/RO membrane filter of aseptic water treatment

  [point]  apply the aseptic membrane filter to purify the bacterial contaminations clear

               a. the 5~1 micron MF filter are applied to purify the bacteria, fungi, pyrogenic

                   organisms clear up to low than ≦100 CFU/ml effectively

               b. the SMF/UF/NF/RO membrane filter are applied to purify all the bacterial

                   contaminations of bacteria, virus, fungi, pyrogenic organisms, endotoxin,

                   pyrogen clear effectively

  [defect]  the filter cartridges and membrane are bacterial growth bed and needed to keep

                filter cartridges and membrane at constant bacteria free, germ free for aseptic

                water process treatment application

3. PAC polymer of aseptic process treatment

  [point]  apply the low dosage for low bacterial contaminations of aseptic treatment and

                high dosage for high bacterial contaminations of aseptic treatment 

  [defect]  a. the PAC polymer are low effect to purify bacteria, virus, fungi, pyrogenic

                    organisms, endotoxin, pyrogen clear

                b. after aseptic purification treatment, The uncoagulated chemical constitutents

                    of PAC polymer are standing in water which are becomed into the extra

                    chemical toxic contaminants  

1. 3M 6406 aerobic count plate: suitable for water, liquid bacterial contamination test

2. 3M 6414 E. Coli count plate: suitable for water, liquid E. coli contamination test

3. MW504 hygiene test: suitable for water, liquid, powder, viscous liquids, surface

                                       bacterial contamination test

4. MW503 E. Coli test: suitable for water, liquid, powder, viscous liquids, surface of

                                      E. Coli contamination test

5. MW572 salmonella test: suitable for water, liquid, powder, viscous liquids, surface of

                                            salmonella contamination test

6. growth promotion test: suitable for culture media growth monitoring and validation

7. QC organisms:  GP-01 Bacillus subtilis          GP-06 Geobacillus stearothermophilus

                     GP-02 Clostridium sporogenes       GP-07 Pseudomonas aeruginosa

                     GP-03 Candida albicans                 GP-08 Staphylococcus aureus

                     GP-04 Aspergillus niger                 GP-09 Escherichia coli

                     GP-05 Micrococcus luteus

Self contained with culture media for simple, convenient, economic, fast and

effective bacteria test of aseptic monitoring and validation  

The standard sterilization treatment, monitoring and validation of steam, dry heat, EO-gas radiation are needed to be confirm the following conditions for correct sterilization treatment, monitoring and validation

1. the bacterial contaminations of the pre-sterilization water, liquids, products should be                disinfected into under≦100 CFU/ml

2. the cooling point of water, liquids, products should be controlled at up to the expected sterilization temperature, dosage, concentration to do the clear expected sterilization time   for at least 8D (99.999999%) sterilization effect and application

3. the sterilization monitoring and validation indicator should be placed at the cooling point of water, liquids, products to monitor and validate the sterilization factor of temperature, dosage, concentration, time and effect right or not

      Sterilization formula:   F  =  D  x  (Log A — Log B)  

The conventional sterilization treatment, monitoring and validation of hospitals, pharmaceutical, medical device, biologicals, cellular products are also to do the clear sterilization treatment, monitoring and validation, but hard to meet the correct aseptic sterilization treatment, monitoring and validation requisite:

[point]  a. apply chemical indicator to monitor and validate or only to check the sterilization

                 temperature, dosage, concentration, time of sterilization unit up to expected

                 sterilization temperature, dosage, concentration and time or not

             b. apply 10⁵ CFU/ml biological indicator to monitor and validate the sterilization

                 effect up to 8D sterilization rate and the residues of aseptic sterilization up to

                 10^-6 CFU/ml or not

[defect] a. the bacterial contaminations of the pre-sterilization water, liquids, products are

                 not disinfected into under≦100 CFU/ml

             b. the indication of monitoring and validation of sterilization temperature, dosage,

                 concentration on the sterilization unit and chemical indicator are not same as the

                 cooling point of sterilization temperature, dosage, concentration

              c. the indication of monitoring and validation of sterilization time are not same as

                  the sterilization functional time under the cooling point of sterilization dosage,

                  temperature, concentration on the clear expected sterilization temperature,

                  dosage, concentration

              d. the aseptic monitoring and validation of 10⁵ CFU/ml biological indicator are

                  only approved the sterilization effect being performed the 5D (99.999%)

                  sterilization rate,

The usually situations and phenmenon of conventional sterilization treatment, monitoring and validation: although applied the long times for more effective sterilization effect, also applied chemical indicator, biological indicator for ensuring the sterilized water, liquids, raw materials, products up to bacteria free, endotoxin free, pyrogen free too, However, the sterilized water, liquids, raw materials and products are still contaminated too, hard to solve and overcome  

1. Steam sterilization validation indicator:   a. spores strips

                                                                      b. self contained biological indicator

                                                                      c. chemical process indicator

                                                                      d. chemical temperature indicator

                                                                      e. sterilization temperature indicator bag

2. Dry heat sterilization validation indicator: a. spores strips

                                                                       b. self contained biological indicator

                                                                       c. chemical temperature indicator      

                                                                       d. sterilization temperature indicator bag

3. EO-gas sterilization validation indicator:  a. spores strips

                                                                       b. self contained biological indicator

                                                                       c. EO-gas test indicator

4. Radiation sterilization validation indicator: a. spores strips

                                                                         b. self contained biological indicator

                                                                         c. radiation chemical indicator

5. Growth promotion test/QC organisms:

     GP-01  Bacillus subtilis                       GP-06  Geobacillus stearothermophilus

     GP-02   Clostridium sporogenes          GP-07  Pseudomonas aeruginosa

     GP-03  Candida albicans                      GP-08  Staphylococcus aureus

     GP-04  Aspergillus niger                      GP-09  Escherichia coli

     GP-05  Micrococcus luteus  

1. Endotoxin test: to monitor and validate the endotoxin contents of gram-negative

                            organisms, inactive carcass

  [effect]  to monitor and validate the endotoxin contents low than 0.2 EU to approve the

                gram-negative organisms and inactive carcass low than 10^-6 CFU/ml or not

  [test kit]  a. conventional gel-cloting LAL test: operation times 2~4 hours

                 b. new endpoint PTS LAL test: operation times 19~32 minutes

2. Pyrogen test: to monitor and validate the pyrogens contents of gram-negative, gram-

                         positive organisms, inactive carcass and other biological pyrogens,

                         including yeast, parasitic and viral pyrogens

  [effect]  to monitor and validate the pyrogens contents low than 0.2 EU to approve all

                the gram-negative gram-positive organisms, inactive carcass and other

                biological organisms and inactive carcass low than 10^-6 CFU/ml or not

  [test kit]  a. conventional rabbit pyrogen test: to monitor and validate the fever response

                 b. in-vitro IPT pyrogen test: to monitor and validate the pyrogens contents  

The pyrogens detection of rabbit test, LAL test and IPT pyrogen test

                            Rabbit test          LAL test                        IPT pyrogen test     

test                   fever reaction      defense mechanism        fever reaction     

gram-negative           +                          +                                       +

gram-positive            +                         –                                       +

fungi                           +                          –                                      +

virus                         +/–¹                     –                                      +

pharmaceutical           +                           +                                        +

biologicals                  +                         +/–²                                   +

medical device           +³                         +/–³                                   +

cellular therapies        –                          +/–³                                   +