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Detection method of aflatoxin in traditional Chinese medicine: 
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Aflatoxin detection protocol in Chinese herbal medicines-HPLC (Pharmacopoeia method)
Aflatoxin harm:
Aflatoxins are toxic metabolites of a class of fungi, such as Aspergillus flavus and Aspergillus parasiticus, which are highly carcinogenic and are found in medicinal materials , grains, nuts, cottonseed, and animal feed-related products. Its toxicity is much higher than that of cyanide, arsenide and organic pesticides, among which B 1 is the most toxic. When a person consumes a large amount, acute poisoning may occur, and acute hepatitis, hemorrhagic necrosis, hepatic steatosis, and bile duct hyperplasia may occur. When the trace is continuously ingested, it can cause chronic poisoning, growth disorders, and cause fibrous lesions, resulting in fibrous tissue hyperplasia.
National attention:
In recent years, food and drug supervision departments at all levels have continuously increased the supervision of Chinese medicine production and circulation, and strive to maintain the overall stability of Chinese medicine quality.
Notice of the State Food and Drug Administration on Regulating the Order of Chinese Medicines to Strictly Investigate Violations of Laws and Regulations (Guo Food and Drug Administration [2012] No. 187, requires strengthening safety indicators for heavy metals and harmful elements, pesticide residues, aflatoxins, etc. The detection and control of the Chinese herbal medicines ensure the quality and safety of the Chinese herbal medicines. In addition, the enterprises are required to have the inspection equipment and capabilities that are compatible with the production varieties, and strictly control the quality.
"National Drug Safety "Twelfth Five-Year Plan", the plan clearly states "to carry out rapid drug testing technology research, build a testing technology sharing platform", "accelerate the application of rapid drug testing technology at the grassroots level, configure rapid testing equipment", Strengthening the requirements of county-level institutions for rapid inspection capability building
The 2010 edition of the Pharmacopoeia Appendix IX V aflatoxin was determined by high performance liquid chromatography, and the additional pharmacopoeia described the method to establish an increased post-column photochemical derivatization method.
As the earliest high-tech enterprise engaged in the detection of mycotoxins in China, Huaan Meike also has a profound exploration and research on aflatoxin detection. Let us share with you the practice of immunoaffinity column-high performance liquid chromatography-post-column photochemistry. Derived and other content.
ã€principle】:
The sample is extracted with methanol-water, the extract is filtered, diluted, and the filtrate is purified by immunoaffinity column chromatography containing aflatoxin-specific antibody . The aflatoxin is cross-linked to the antibody in the chromatographic medium, and the antibody is against Aspergillus flavus. The toxins B 1 , B 2 , G 1 and G 2 have specificity, and the impurities on the immunoaffinity column are removed by water, eluted with methanol through an immunoaffinity column, and then derivatized by a photochemical derivatization column to improve detection accuracy. Sex.
[Main reagents and consumables]:
ToxinFast ® air control operating frame : air pump
ToxinFast ® aflatoxin immunoaffinity column 1ml or 3ml
ToxinFast ® Photochemical Derivatives
ToxinFast ® glass microfiber filter paper : diameter 11cm, aperture 1.5μm
ToxinFast ® glass test tube : 12mm in diameter, 75mm in length, no fluorescence
ToxinFast ® syringe : 10ml, 20ml
[Detection steps]:
Sample pulverization - extraction - eluent injection, affinity column purification - chemical derivation - detection of values ​​(all of these steps are known in the liquid phase, here is a brief mention, mainly why the gas is used The control frame and chemical derivative are explained below!)
Instructions for use of the air control operating frame:
After the sample is pulverized, it needs to be cleaned by the affinity column, which is how to facilitate the operation of the affinity column. The air control operation frame can effectively improve the working efficiency of this link (very convenient operation), as shown in the figure:
1 Connect the assembled air control operation frame, connect the immunoaffinity column and the syringe syringe to the air control operation frame as shown in the figure;
2 loading the treated solution;
3 adjust the air pump knob to control the flow rate, so that the sample liquid flows out at a speed of 1-2d/s;
4, the sample solution is completely drained, washed twice with deionized water or distilled water at a flow rate of 2-3d / s, each time 10ml;
5. After the liquid is completely drained, place the vial under the affinity column and load 1 ml of methanol to flow out at 1 d/s until it is completely drained.
The above steps are the necessary steps to detect aflatoxin in traditional Chinese medicine by immunoaffinity column-high performance liquid phase method. If the air-controlled operating frame is used, the affinity column can be well fixed and operated at a controlled flow rate, which is efficient. It also minimizes operational errors.
Chinese An Maike ToxinFast ® pneumatic operating frame having six controllable tributaries can effectively detect a plurality of samples simultaneously, is also applicable to
1ml and 3ml immunological affinity columns of various specifications.
Why use a photochemical derivative?
Due to its strong UV absorption and fluorescence-generating properties, Aspergillus flavus can be subjected to HPLC-ultraviolet or fluorescence detection, but at the same time, B 1 and G 1 in aflatoxin are prone to fluorescence quenching due to their presence in the aqueous mobile phase. Therefore, in many experiments, a derivative method is used to determine aflatoxin.
There are also many methods of derivatization: pre-column, post-column, and online. Due to the obvious advantages of post-column photochemical derivatization, it is now supplemented as a pharmacopoeia detection method. The main advantage lies in ( no chemical reagents, no No harm to personnel; no need for direct operation by personnel, avoiding errors; no need to worry about derivative liquid corrosion detector )
After reading the above picture, you can definitely feel the effect of online photochemical derivation. I hope this sharing can provide a little help for everyone to do aflatoxin detection.