Prosecutors say Deborah Leggio pleaded not guilty Tuesday at her arraignment in Suffolk County Court in Riverhead to second-degree grand larceny.
Leggio, who now lives in Tracy, Calif., allegedly used a Red Cross debit card to pay for personal items like nail treatments, Weight Watchers membership and tuition for her children's education.
Leggio is alleged to have taken the money between August 2005 and November 2009. An audit later uncovered the missing funds.
Bail was set at $17,500. Her attorney did not immediately return a call for comment.
Suffolk Red Cross spokesman Craig Cooper says the Long Island agency cooperated with the DA's investigation.