Prospects for holding a May 1 referendum on legalizing some forms of gambling in the District of Columbia suffered a serious setback yesterday in the D.C. City Council.

William R. Spaulding (D-Ward 5), chairman of the council's Government Operations Committee, announced that he had withdrawn from consideration legislation calling for the referendum scheduled to be heard at a committee meeting today.

The measure, proposed by the Citizens Study Commission that submitted a report to the council last May, would create a city-run lottery, authorize pari-mutuel betting on sports events and legalize some forms of charitable gambling, such as raffles and bingo.

Spaulding's announcement drew immediate fire from Council Chairman Arrington L. Dixon, chief sponsor of the measure calling for the referendum.

"The people deserve to be heard on this issue and this resolution deserves to be voted up or down in the committee... and in the council," Dixon said.

The Government Operations Committee had lined up, 3 to 2, against the gambling measure. Dixon and some others have contended that the referendum plan would win support from a majority of the full 12-member council.

Spaulding said most of the lobbying for the gambling measure had been done by Jerry Cooper, a member of the study commission that recommended it. "But every citizens association and PTA president down the line has asked me to vote 'no,'" he told reporters.

At its meeting yesterday, the council came close to challenging the right of Congress to insist upon having council-passed legislation conform to congressional policies, but finally decided to go along with Congress.

The issue arose when the Senate subcommittee that deals with D.C. governmental matters objected -- on highly technical legal grounds -- to two provisions in the city's new personnel law, scheduled to go into effect next week. They dealt with preserving suburban residency rights of all current D.C. employes and with maintaining Hatch Act provisions restricting partisan political activity.

Initially, a three-member bloc composed of Hilda Mason (Statehood-At Large), Jerry A. Moore (R-At Large) and Wilhelmina Rolark (D-Ward 8) managed to prevent the declaration of an emergency needed to amend the measure. Rolark, later convinced that the position would harm the city, switched her vote and the measure was passed.

On other matters, the council voted to approve the appointments of Ruth O. Robinson as council secretary, James M. Christian as general counsel and mayoral aides Douglas N. Schneider Jr. and Gladys Mack as alternate directors of the Metro transit system.