US 11,873,535 B2
Authentication of botanical DNA isolated from dietary supplements
Zhengfei Lu, Alhambra, CA (US); Silva Babajanian, Porter Ranch, CA (US); Yanjun Zhang, Reseda, CA (US); Peter Chang, Newbury Park, CA (US); Gary Swanson, Mission Viejo, CA (US); and Maria Rubinsky, Redondo Beach, CA (US)
Assigned to HERBALIFE INTERNATIONAL OF AMERICA, INC., Los Angeles, CA (US)
Filed by Herbalife International of America, Inc., Los Angeles, CA (US)
Filed on Jul. 27, 2018, as Appl. No. 16/048,091.
Claims priority of provisional application 62/562,101, filed on Sep. 22, 2017.
Claims priority of provisional application 62/538,423, filed on Jul. 28, 2017.
Prior Publication US 2019/0032153 A1, Jan. 31, 2019
Int. Cl. C12Q 1/6895 (2018.01); C12Q 1/6855 (2018.01)
CPC C12Q 1/6895 (2013.01) [C12Q 1/6855 (2013.01)] 18 Claims
 
1. A method of authenticating sterilized chamomile material in a botanical dietary supplement, the method comprising:
isolating genomic DNA from a botanical dietary supplement comprising German chamomile, feverfew, Roman chamomile, and Chinese chamomile, wherein the geomic chamomile DNA isolated from the botanical dietary supplement comprises chamomile DNA fragments, wherein at least some of the chamomile DNA fragments are less than 220 base pairs, wherein the genomic chamomile DNA isolated from the botanical dietary supplement is degraded and undetectable using DNA barcoding, and wherein the chamomile DNA fragments are present in an amount of 1 pg to 1 ng;
ligating adapters to the chamomile DNA fragments to generate adapter-ligated DNA fragments;
amplifying the adapter-ligated DNA fragments using 8-12 cycles of polymerase chain reaction (PCR);
detecting the chamomile DNA fragments;
determining a size range of the chamomile DNA fragments;
hybridizing primers to a target nucleic acid sequence in the chamomile DNA fragments, wherein the primers comprise a species-specific primer set selected from:
a primer comprising a nucleic acid sequence as defined in SEQ ID NO: 1 and a primer comprising a nucleic acid sequence as defined in SEQ ID NO: 2;
a primer comprising a nucleic acid sequence as defined in SEQ ID NO: 3 and a primer comprising a nucleic acid sequence as defined in SEQ ID NO: 4;
a primer comprising a nucleic acid sequence as defined in SEQ ID NO: 5 and a primer comprising a nucleic acid sequence as defined in SEQ ID NO: 6; and
a primer comprising a nucleic acid sequence as defined in SEQ ID NO: 7 and a primer comprising a nucleic acid sequence as defined in SEQ ID NO: 8, and
amplifying the target nucleic acid sequence.