The sanctions against 10 companies and six individuals are created to disrupt the economic ties that have allowed Pyongyang to continue funding its missile and nuclear program despite strict United Nations sanctions prohibiting it.
The Treasury Department, which oversees United States sanctions programs, said those targeted by the sanctions had helped people known to support the North's nuclear and ballistic missile programs, or had traded in North Korean energy, exploited its workers or acted as a conduit to the worldwide financial system.
US sanctions, even though they are imposed by just one country, have an outsize influence because most worldwide banking is conducted in USA dollars.
The 16 sanctioned entities are doing business with previously sanctioned companies and people, work in North Korea's energy sector, help move North Korean workers to jobs overseas and allow the government to evade sanctions and access worldwide financial markets, Treasury said.
The Office of Foreign Assets Control designated 10 "third-country" entities, including six Chinese-owned, one Russian, one North Korean and two based in Singapore.
The move follows a war of words between Washington and Pyongyang over recent missile tests that show North Korea is capable of reaching the USA mainland.
Though U.S. military officials and President Donald Trump have said that the United States is prepared to take some sort of military action against North Korea if provoked, Secretary of State Rex Tillerson has pointedly called for Pyongyang to negotiate and said repeatedly that the United States does not seek regime change.
He added: "It is unacceptable for individuals and companies in China, Russia and elsewhere to enable North Korea to generate income used to develop weapons of mass destruction and destabilize the region".
The U.S. Justice Department is also seeking forfeitures of $11 million from firms that allegedly laundered money for North Korean banks. Ltd by the U.S. Attorney for the District of Columbia, represent two of the largest North Korea-related seizures the government has ever pursued.
North Korea uses the revenue from those exports to fund its missile programs, the Treasury Department says.
In total, the US has blacklisted 23 entities and 22 individuals linked to North Korea since the launch of the Trump administration in January.
China reacted with irritation, saying Washington should "immediately correct its mistake" of imposing unilateral sanctions on Chinese companies and individuals to avoid damaging bilateral cooperation.
The latest steps stopped short of targeting Chinese financial institutions dealing with North Korea, a step that would have greatly angered Beijing.
South Korea's foreign ministry, meanwhile, welcomed the announcement, saying it hopes the sanctions will boost the allies' joint efforts to denuclearize the North.