The helpline opened on Monday following allegations of sexual misconduct which have emerged at the Scottish Parliament and Westminster.
A Scottish Parliament spokesman said four calls had been received as of 5pm on Wednesday but declined to comment further.
The phone line was set up to provide advice and support to workers at Holyrood and in constituency and regional offices and is open from 9am to 5pm Monday to Friday.
In an email to staff announcing the new phone line, the parliament’s chief executive Sir Paul Grice said his priority was to “ensure that anyone suffering from harassment has the advice and support that they need”.
He added: “We want to foster a positive culture where everyone is treated with respect and everyone feels confident to speak out about unacceptable behaviour.”
Last week Mark McDonald resigned as the Scottish Government’s minister for childcare and early years over past actions he said had been deemed “inappropriate’’.
Meanwhile, Central Scotland MSP Monica Lennon revealed she was ‘’groped’’ at a social function in 2013 by a senior member of the Labour Party.
Holyrood bosses also said they had received a complaint from a worker regarding SNP backbencher Willie Coffey, alleging he used ‘’inappropriate language’’ and directed ‘’unsolicited attention’’ towards a female administrative assistant.
A spokesman for the MSP said he did not ‘’recognise the claims that are being made about his behaviour’’ and a Scottish Parliament spokeswoman said the matter had been ‘’dealt with appropriately”.