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Hyaluronic acid - Hyasis® FAQs

General questions

What is Hyasis?

Hyasis is Novozymes Bacillus subtilis-derived sodium hyaluronate. Hyasis is a premium ingredient with applications in the medical device and biopharmaceutical industries. Hyasis comes in two grades: topical and parenteral.
 

What is Bacillus subtilis?

Bacillus subtilis is the bacterial strain used in the production of Hyasis. It has been modified to express the hasA gene from Streptococcus equisimilis which encodes for the enzyme hyaluronan synthase. Artificial operons were assembled and tested, all of which contain the hasA gene along with other gene-encoding enzymes involved in the synthesis of the UDP precursor sugars required for hyaluronan synthesis.
 

Why produce Hyasis using Bacillus subtilis?

Bacillus subtilis is a well-characterized bacterial strain used for the production of a wide range of commercially available products. Bacillus subtilis has a number of unique advantages, and Novozymes possesses great expertise in handling Bacillus subtilis. Hyasis is produced without any animal-derived raw materials and directly extrudes HA into the fermentation medium. This explains the uniqueness of Novozymes manufacturing process: water-based and devoid of any organic solvents this process allows the production of a highly pure and consistent sodium hyaluronate material.
 

Sampling

How can I get a sample of Hyasis?

You can obtain a sample of Hyasis by clicking the contact us link at the top of the page. If you are interested in an experimental grade sodium hyaluronate material, we will ask you to complete a sampling agreement. This sampling agreement typically discloses the scientific field of use of the sample and sample quantity. It is signed by both you and Novozymes, and each party receives a copy of it. Once the agreement is in place, the experimental sample will be sent to you.
 

Handling and formulation

How should I store Hyasis?

In the short-term (during transport, etc.) unopened Hyasis can be stored at ambient temperature. In the long-term unopened Hyasis should be stored in a cool (2–8 °C) and dry place. Due to the well-known high hygroscopicity of sodium hyaluronate, we recommend storing opened Hyasis tightly sealed in a cool (2–8 °C) and dry place.
 

Can I freeze solutions containing Hyasis?

Our results show that the molecular weight of Hyasis remains unchanged when solutions containing Hyasis are frozen down to temperatures below -196 °C (temperature of liquid nitrogen).
 

Should I be aware of any particular precaution when formulating Hyasis?

Sodium hyaluronate is known to be sensitive to
• strong acidic and basic conditions (pH outside the range 5.0–9.0)
• strong oxidative conditions
• high temperatures in solution (> 60 °C for extended periods of time)
• sterilization by irradiation
• ultrasound
For further information on material handling, feel free to consult our material handling recommendation sheet.
 

How is Hyasis best solubilized in aqueous media?

Sodium hyaluronate is best solubilized by progressively adding it to the solution medium into the vortex created by the agitation equipment. The time required to dissolve sodium hyaluronate mainly depends on its molecular weight and concentration but also on the temperature and type of stirring device used. We advise against using ultrasonication to accelerate the solubilization process. Solubilization can however be conducted above room temperature (< 60 °C for maximum 6 hours) in order to reduce dissolving time.
 

Is Hyasis soluble in organic solvents?

Sodium hyaluronate is a highly hydrophilic molecule which, in principle, is not soluble in organic solvents. However, it can be solubilized in mixtures of water and organic solvents such as aqueous ethanol (maximum 70% v/v).
 

Which methods can be applied to sterilize solutions containing Hyasis?

Applicable methods to sterilize sodium hyaluronate solutions are filtration and autoclaving.
Filtration through a 0.22-μm porous filter has been shown to be feasible at production scale for buffered Hyasis solutions with a molecular weight up to 1 MDa and a concentration up to 1% w/v.
Autoclaving is also applicable although it may lead to some polymer degradation. If autoclaving is selected as sterilization method, we recommended minimizing the presence of Cu and Fe ionic species since these are known to catalyze depolymerization.
We advise against using irradiation since this technique leads to extensive depolymerization of sodium hyaluronate.
 

Under which conditions does Hyasis degrade?

In nature hyaluronic acid (the acidic residue of sodium hyaluronate) is degraded enzymatically by the action of hyaluronidase or other lyases. Sodium hyaluronate is also degraded chemically when subjected to strong acidic, basic, or oxidative conditions. Exposure to an elevated temperature, irradiation and/or ultrasound can also lead to polymer degradation.
 
 

Specifications

How much endotoxin does Hyasis contain?

As Bacillus subtilis does not contain endotoxins, the Hyasis manufacturing process can deliver material with very low endotoxin levels. All our grades of Hyasis comply, as a minimum, with the requirements of the European Pharmacopoeia. Please contact us if you have any questions regarding specific levels of endotoxin for your application by clicking the contact us link at the top of the page.
 

Customer technical support

What technical and application data can you share on Hyasis?

We can share data on the fundamental properties of sodium hyaluronate solutions, formulation, stability under various conditions and treatments, and compatibility with excipients and active pharmaceutical ingredients. Please visit www.biopharma.novozymes.com or clicking the contact us link at the top of the page.
 

Does Hyasis contain other metal counterions than sodium?

Hyasis primarily contains sodium counterions. However, it also contains very low amounts of calcium and magnesium ionic species. Please contact us by clicking the contact us link at the top of the page if you have any questions regarding specific levels of metal ions for your application.
 

Regulatory affairs

According to which standards is Hyasis produced?

The parenteral grade of Hyasis is produced according to ICH Q7. ICH Q7 is the Good Manufacturing Practice (GMP) for active pharmaceutical ingredients (APIs). The topical grade of Hyasis is produced according to the principles of IPEC-PQG GMP.
 

Which type of Drug Master File (DMF) is Hyasis associated with? Can I have access to this information?

A Drug Master File Type IV (used for excipients) will be submitted to the US/FDA for Hyasis 850T (topical grade). A Drug Master File Type II (used for APIs) will be submitted to the US/FDA for Hyasis 850P (parenteral grade). Submission of these two DMFs will take place as soon as the needed documentation is available. An Open Part of each DMF can be made available under a Confidential Disclosure Agreement (CDA) and a Letter of Authorization to reference the DMFs can be issued to customers, if needed.
 

Is Hyasis approved by regulatory bodies?

Pharmaceutical ingredients are not approved in their own right. However Hyasis is supported by a comprehensive regulatory support package and is currently used in several FDA/EU regulated products.

Does Hyasis possess the Generally Recognized as Safe (GRAS) status?

GRAS is a US/FDA designation attributed to a substance considered safe by experts when added to food. Since Hyasis is not intended to be added to food it does not have the GRAS status.
Hyasis is produced by fermentation of the safe bacterial strain Bacillus subtilis. Several enzymes manufactured by Novozymes possess the GRAS status. Furthermore, Bacillus subtilis is included in the list of Quality Presumption of Safety recommended by the European Food Safety Authority.
 

How can Novozymes support the registration of my product?

Novozymes can support the registration of your product by giving you access to our comprehensive regulatory support package. Please contact us by clicking the contact us link at the top of the page.
 
Last updated Janurary 2013
We strive to provide accurate and useful support, however, the information contained in this document is provided solely as guidance to the intended recipient. Furthermore, laws, regulations and/or third party rights may prevent the recipient from using the information herein in a given manner. Thus, without separate, written agreement between the recipient and Novozymes to a different effect, the information contained herein is provided “as is” and Novozymes makes no representation or warranty whatsoever with regards to said information, hereunder any warranty as to the accuracy, fitness for a particular purpose, non-infringement of intellectual property rights or regulatory/legal compliance.

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